diff --git a/.dockerignore b/.dockerignore index e7590d75..fa1076d6 100644 --- a/.dockerignore +++ b/.dockerignore @@ -5,3 +5,4 @@ main/src/tests **/.ipynb_checkpoints/ README.md LICENSE +/.venv \ No newline at end of file diff --git a/.github/workflows/continuous_integration.yml b/.github/workflows/continuous_integration.yml new file mode 100644 index 00000000..8109f213 --- /dev/null +++ b/.github/workflows/continuous_integration.yml @@ -0,0 +1,84 @@ +name: Continuous Integration +on: + push: + branches: + - main + pull_request: + workflow_dispatch: + +jobs: + format: + runs-on: ubuntu-latest + steps: + - name: Checkout + uses: actions/checkout@v4 + + - name: Install uv + uses: astral-sh/setup-uv@v3 + + - name: Create Virtual Environment + run: uv venv + + - name: Install Jupyter + run: uv pip install jupyter nbconvert + + - name: Strip Notebook Output + run: uv run jupyter nbconvert --clear-output --inplace "main/COMO.ipynb" + + - name: Format Python Imports + uses: astral-sh/ruff-action@v1 + with: + args: "check --fix --select I" + + - name: Format code + uses: astral-sh/ruff-action@v1 + with: + args: "format" + + - name: Format Notebook + uses: astral-sh/ruff-action@v1 + with: + args: "format main/COMO.ipynb" + + - name: Commit Changes + uses: stefanzweifel/git-auto-commit-action@v5 + with: + commit_message: "style: format code, Jupyter Notebook(s), and Python imports with `ruff`" + file_pattern: "main/como/*.py" + + + lint: + runs-on: ubuntu-latest + steps: + - name: Checkout + uses: actions/checkout@v4 + + - name: Check Lint + uses: astral-sh/ruff-action@v1 + with: + args: "check --no-fix --verbose" + + test: + runs-on: ubuntu-latest + strategy: + matrix: + python-version: [ "3.10" ] + steps: + - name: Checkout + uses: actions/checkout@v4 + + - name: Install uv + uses: astral-sh/setup-uv@v4 + with: + enable-cache: "true" + cache-suffix: "${{ matrix.python-version }}" + cache-dependency-glob: 'uv.lock' + + - name: Install Dependencies + run: uv sync --python "${{ matrix.python-version }}" --all-extras --dev + + - name: Run tests + run: uv run --python "${{ matrix.python-version }}" pytest --cov --junitxml=junit.xml -o junit_family=legacy + + - name: Cache Clear + run: uv cache prune --ci diff --git a/.github/workflows/unit_tests.yml b/.github/workflows/unit_tests.yml deleted file mode 100644 index e40acef3..00000000 --- a/.github/workflows/unit_tests.yml +++ /dev/null @@ -1,40 +0,0 @@ -name: Unit Tests -on: - workflow_dispatch: - pull_request: - types: - - opened - - reopened - - synchronize - - -jobs: - unit-tests: - runs-on: ubuntu-latest - - # Using a matrix lets us define additional versions of Python to test against - strategy: - matrix: - python-version: [ "3.10" ] - - steps: - - name: Checkout Repository - uses: actions/checkout@v4 - - - name: Install Conda environment - uses: mamba-org/setup-micromamba@v2 - with: - environment-file: main/tests/environment.yaml - environment-name: como_tests - micromamba-version: "latest" - init-shell: bash - cache-environment: true - cache-downloads: true - post-cleanup: "none" - create-args: >- - python=${{ matrix.python-version }} - - - name: Run tests - run: "cd main && python -m pytest" - shell: micromamba-shell {0} - diff --git a/.gitignore b/.gitignore index f0c49976..6123cff0 100644 --- a/.gitignore +++ b/.gitignore @@ -12,14 +12,13 @@ docs/_site docs/Gemfile.lock # COMO-specific files +.hypothesis **/*.Rout main/logs/ -main/data/results/ -main/data/config_sheets/*_data_inputs_auto.xlsx main/data/gene_info.csv main/data/Repurposing_Hub_Preproc.tsv -main/microarray.db -main/data/config_sheets/* -main/data/GSE*_RAW -main/data/gpl*entrez.csv -main/src/microarray.db \ No newline at end of file +main/data/results/ +main/data/config_sheets + +# COMO data +main/data/data_matrices diff --git a/.pre-commit-config.yaml b/.pre-commit-config.yaml new file mode 100644 index 00000000..16d682c0 --- /dev/null +++ b/.pre-commit-config.yaml @@ -0,0 +1,8 @@ +# See https://pre-commit.com for more information +# See https://pre-commit.com/hooks.html for more hooks +repos: +- repo: https://github.com/opensource-nepal/commitlint + rev: v1.2.0 + hooks: + - id: commitlint + name: Commit Lint diff --git a/Dockerfile b/Dockerfile index 8825d49b..850064f2 100644 --- a/Dockerfile +++ b/Dockerfile @@ -1,27 +1,12 @@ -FROM jupyter/r-notebook:latest as builder +FROM python:3.10 AS app -COPY environment.yaml "${HOME}/environment.yaml" -COPY --chown=1000:100 main "${HOME}/main" - -# Install python-related items -# Remove "python" from the pinned file so we can install our own version -RUN sed -i '/^python/d' /opt/conda/conda-meta/pinned && \ - echo "auto_activate_base: true" >> "${HOME}/.condarc" && \ - jupyter trust "${HOME}/main/COMO.ipynb" && \ - echo "c.ServerApp.ip = '0.0.0.0'" >> "${HOME}/.jupyter/jupyter_notebook_config.py" && \ - echo "c.ServerApp.root_dir = '${HOME}/main'" >> "${HOME}/.jupyter/jupyter_notebook_config.py" && \ - echo "c.ServerApp.token = ''" >> "${HOME}/.jupyter/jupyter_notebook_config.py" && \ - echo "c.ServerApp.password = ''" >> "${HOME}/.jupyter/jupyter_notebook_config.py" && \ - mamba config --quiet --add channels conda-forge && \ - mamba config --quiet --add channels bioconda && \ - mamba config --quiet --add channels r && \ - rm -rf "${HOME}/main/tests" # Tests not required for running COMO - -# Update base environment -RUN mamba env update --name=base --file="${HOME}/environment.yaml" && \ - R -e "devtools::install_github('babessell1/zFPKM')" && \ - pip cache purge && \ - mamba clean --all --yes +WORKDIR /app +ENV PATH="/app/.venv/bin:$PATH" +COPY --from=ghcr.io/astral-sh/uv:latest /uv /uvx /bin/ +COPY --chown=1000:100 main /app/main/ +COPY --chown=1000:100 pyproject.toml /app/pyproject.toml +RUN uv sync && uv pip install jupyterlab EXPOSE 8888 -VOLUME /home/joyvan/main/data/local_files +VOLUME "/app/main/data/local_files" +CMD ["jupyter", "lab", "--allow-root", "--no-browser", "--ip=0.0.0.0", "--port=8888", "--notebook-dir=/app/main", "--NotebookApp.token=''", "--NotebookApp.password=''"] diff --git a/README.md b/README.md index 20d00b76..b00162bc 100644 --- a/README.md +++ b/README.md @@ -3,6 +3,8 @@ ![GitHub release (with filter)](https://img.shields.io/github/v/release/HelikarLab/COMO?filter=v*-master&style=for-the-badge&color=blue) ![GitHub Workflow Status (with event)](https://img.shields.io/github/actions/workflow/status/HelikarLab/COMO/unit_tests.yml?style=for-the-badge&logo=pytest&logoColor=white&label=Tests) ![GitHub Workflow Status (with event)](https://img.shields.io/github/actions/workflow/status/HelikarLab/COMO/container_build.yml?style=for-the-badge&logo=docker&logoColor=white&label=Docker%20Build) +[![pre-commit](https://img.shields.io/badge/pre--commit-enabled-brightgreen?logo=pre-commit)](https://github.com/pre-commit/pre-commit) + # Setting up COMO diff --git a/docker-compose.yml b/docker-compose.yml new file mode 100644 index 00000000..310b864f --- /dev/null +++ b/docker-compose.yml @@ -0,0 +1,9 @@ +services: + como: + image: ghcr.io/helikarlab/como:latest + container_name: como + ports: + - "8888:8888" + volumes: + # Map "[LOCAL_DIRECTORY]/local_files to /app/main/data/local_files + - "$(pwd)/local_files:/app/main/data/local_files" \ No newline at end of file diff --git a/main/COMO.ipynb b/main/COMO.ipynb index ebed25a0..8178cd67 100644 --- a/main/COMO.ipynb +++ b/main/COMO.ipynb @@ -235,29 +235,22 @@ ] }, { + "metadata": {}, "cell_type": "code", - "execution_count": null, - "metadata": { - "pycharm": { - "is_executing": true - }, - "tags": [] - }, "outputs": [], + "execution_count": null, "source": [ "context_names = \"naiveB\"\n", - "gene_format = \"Ensembl\" # accepts \"Entrez\", \"Ensembl\", and \"Symbol\"\n", "taxon_id = \"human\" # accepts integer (bioDBnet taxon id) or \"human\" or \"mouse\"\n", - "preprocess_mode = \"create-matrix\" # \"create-matrix\" or \"provide-matrix\"\n", + "preprocess_mode = \"create\" # \"create\" or \"provide\"\n", "\n", "# fmt: off\n", "cmd = \" \".join(\n", " [\n", " \"python3\", \"como/rnaseq_preprocess.py\",\n", - " \"--context-names\", f\"{context_names}\",\n", - " \"--gene-format\", f\"{gene_format}\",\n", - " \"--taxon-id\", f\"{taxon_id}\",\n", - " f\"--{preprocess_mode}\",\n", + " \"--context-names\", context_names,\n", + " \"--taxon-id\", taxon_id,\n", + " \"--mode\", preprocess_mode,\n", " ]\n", ")\n", "# fmt: on\n", @@ -267,11 +260,7 @@ }, { "cell_type": "markdown", - "metadata": { - "jupyter": { - "outputs_hidden": false - } - }, + "metadata": {}, "source": [ "\n", "## Identification of Gene Activity in Transcriptomic and Proteomic Datasets\n", @@ -340,12 +329,10 @@ ] }, { + "metadata": {}, "cell_type": "code", - "execution_count": null, - "metadata": { - "tags": [] - }, "outputs": [], + "execution_count": null, "source": [ "# step 2.2 RNA-seq Analysis for Total RNA-seq library preparation\n", "\n", @@ -355,22 +342,22 @@ "rep_ratio_h = 1.0\n", "group_ratio_h = 1.0\n", "technique = \"zFPKM\"\n", - "quantile = 50\n", - "min_zfpkm = -3\n", - "prep_method = \"total\"\n", + "minimum_cutoff = -3\n", + "taxon_id = \"human\"\n", "\n", "# fmt: off\n", "cmd = \" \".join(\n", " [\n", " \"python3\", \"como/rnaseq_gen.py\",\n", - " \"--config-file\", f\"{trnaseq_config_file}\",\n", - " \"--replicate-ratio\", f\"{rep_ratio}\",\n", - " \"--batch-ratio\", f\"{group_ratio}\",\n", - " \"--high-replicate-ratio\", f\"{rep_ratio_h}\",\n", - " \"--high-batch-ratio\", f\"{group_ratio_h}\",\n", + " \"--config-file\", trnaseq_config_file,\n", + " \"--replicate-ratio\", str(rep_ratio),\n", + " \"--batch-ratio\", str(group_ratio),\n", + " \"--high-replicate-ratio\", str(rep_ratio_h),\n", + " \"--high-batch-ratio\", str(group_ratio_h),\n", + " \"--minimum-cutoff\", str(minimum_cutoff),\n", " \"--filt-technique\", f\"{technique}\",\n", - " \"--min-zfpkm\", f\"{min_zfpkm}\",\n", - " \"--library-prep\", f\"{prep_method}\",\n", + " \"--library-prep\", \"total\",\n", + " \"--taxon-id\", taxon_id\n", " ]\n", ")\n", "# fmt: on\n", @@ -398,12 +385,10 @@ ] }, { + "metadata": {}, "cell_type": "code", - "execution_count": null, - "metadata": { - "tags": [] - }, "outputs": [], + "execution_count": null, "source": [ "mrnaseq_config_file = \"mrnaseq_data_inputs_auto.xlsx\"\n", "rep_ratio = 0.75\n", @@ -411,23 +396,76 @@ "rep_ratio_h = 1.0\n", "group_ratio_h = 1.0\n", "technique = \"zfpkm\"\n", - "quantile = 50\n", - "min_zfpkm = -3\n", - "prep_method = \"mrna\"\n", + "minimum_cutoff = -3\n", + "taxon_id = \"human\"\n", "\n", "# fmt: off\n", "cmd = \" \".join(\n", " [\n", " \"python3\", \"como/rnaseq_gen.py\",\n", - " \"--config-file\", f\"{mrnaseq_config_file}\",\n", - " \"--replicate-ratio\", f\"{rep_ratio}\",\n", - " \"--batch-ratio\", f\"{group_ratio}\",\n", - " \"--high-replicate-ratio\", f\"{rep_ratio_h}\",\n", - " \"--high-batch-ratio\", f\"{group_ratio_h}\",\n", + " \"--config-file\", mrnaseq_config_file,\n", + " \"--replicate-ratio\", str(rep_ratio),\n", + " \"--batch-ratio\", str(group_ratio),\n", + " \"--high-replicate-ratio\", str(rep_ratio_h),\n", + " \"--high-batch-ratio\", str(group_ratio_h),\n", + " \"--minimum-cutoff\", str(minimum_cutoff),\n", " \"--filt-technique\", f\"{technique}\",\n", - " \"--min-zfpkm\", f\"{min_zfpkm}\",\n", - " \"--quantile\", f\"{quantile}\",\n", - " \"--library-prep\", f\"{prep_method}\",\n", + " \"--library-prep\", \"mrna\",\n", + " \"--taxon-id\", taxon_id\n", + " ]\n", + ")\n", + "# fmt: on\n", + "\n", + "!{cmd}" + ] + }, + { + "cell_type": "markdown", + "metadata": {}, + "source": [ + "## Single-Cell RNA Sequencing Generation\n", + "These parameters are identical to the ones listed for [total RNA sequencing](#Total-RNA-Sequencing-Generation), but they are listed again here for ease of reference\n", + "\n", + "### Parameters\n", + "- `scrnaseq_config_file`: The configuration filename for total RNA. This file is found under the `data/config_sheets` folder\n", + "- `rep_ratio`: The proportion of replicates before a gene is considered \"active\" in a study\n", + "- `group_ratio`: The proportion of studies with expression required for a gene to be considered \"active\"\n", + "- `rep_ratio_h`: The proportion of replicates that must express a gene before that gene is considered \"high-confidience\"\n", + "- `group_ratio_h`: The proportion of studies that must express a gene before that gene is considered \"high-confidence\"\n", + "- `technique`: The only option offered for single-cell RNA sequencing us \"umi\"\n", + "- `quantile`: The cutoff Transcripts-Per-Million quantile for filtering\n", + "- `min_zfpkm`: The cutoff for Counts-Per-Million filtering\n", + "- `prep_method`: The library method used for preparation. Options are: `\"total\"`, `\"mRNA\"`, or `\"scrna\"`,\n" + ] + }, + { + "metadata": {}, + "cell_type": "code", + "outputs": [], + "execution_count": null, + "source": [ + "scrnaseq_config_file = \"scrnaseq_data_inputs_auto.xlsx\"\n", + "rep_ratio = 0.75\n", + "group_ratio = 0.75\n", + "rep_ratio_h = 1.0\n", + "group_ratio_h = 1.0\n", + "quantile = 50\n", + "minimum_cutoff = -3\n", + "taxon_id = \"human\"\n", + "\n", + "# fmt: off\n", + "cmd = \" \".join(\n", + " [\n", + " \"python3\", \"como/rnaseq_gen.py\",\n", + " \"--config-file\", scrnaseq_config_file,\n", + " \"--replicate-ratio\", str(rep_ratio),\n", + " \"--batch-ratio\", str(group_ratio),\n", + " \"--high-replicate-ratio\", str(rep_ratio_h),\n", + " \"--high-batch-ratio\", str(group_ratio_h),\n", + " \"--minimum-cutoff\", str(minimum_cutoff),\n", + " \"--filt-technique\", \"umi\",\n", + " \"--library-prep\", \"scrna\",\n", + " \"--taxon-id\", taxon_id\n", " ]\n", ")\n", "# fmt: on\n", @@ -454,9 +492,7 @@ { "cell_type": "code", "execution_count": null, - "metadata": { - "tags": [] - }, + "metadata": {}, "outputs": [], "source": [ "proteomics_config_file = \"proteomics_data_inputs_paper.xlsx\"\n", @@ -470,12 +506,12 @@ "cmd = \" \".join(\n", " [\n", " \"python3\", \"como/proteomics_gen.py\",\n", - " \"--config-file\", f\"{proteomics_config_file}\",\n", - " \"--replicate-ratio\", f\"{rep_ratio}\",\n", - " \"--high-replicate-ratio\", f\"{high_rep_ratio}\",\n", - " \"--batch-ratio\", f\"{batch_ratio}\",\n", - " \"--high-batch-ratio\", f\"{high_batch_ratio}\",\n", - " \"--quantile\", f\"{quantile}\",\n", + " \"--config-file\", proteomics_config_file,\n", + " \"--replicate-ratio\", str(rep_ratio),\n", + " \"--high-replicate-ratio\", str(high_rep_ratio),\n", + " \"--batch-ratio\", str(batch_ratio),\n", + " \"--high-batch-ratio\", str(high_batch_ratio),\n", + " \"--quantile\", str(quantile),\n", " ]\n", ")\n", "# fmt: on\n", @@ -531,19 +567,19 @@ "cmd = \" \".join(\n", " [\n", " \"python3\", \"como/cluster_rnaseq.py\",\n", - " \"--context-names\", f\"{context_names}\",\n", - " \"--filt-technique\", f\"{filt_technique}\",\n", - " \"--cluster-algorithm\", f\"{cluster_algorithm}\",\n", - " \"--label\", f\"{label}\",\n", - " \"--min-dist\", f\"{min_dist}\",\n", - " \"--replicate-ratio\", f\"{replicate_ratio}\",\n", - " \"--batch-ratio\", f\"{batch_ratio}\",\n", - " \"--n-neighbors-rep\", f\"{n_neighbors_rep}\",\n", - " \"--n-neighbors-batch\", f\"{n_neighbors_batch}\",\n", - " \"--n-neighbors-context\", f\"{n_neighbors_context}\",\n", - " \"--min-count\", f\"{min_count}\",\n", - " \"--quantile\", f\"{quantile}\",\n", - " \"--seed\", f\"{seed}\",\n", + " \"--context-names\", context_names,\n", + " \"--filt-technique\", filt_technique,\n", + " \"--cluster-algorithm\", cluster_algorithm,\n", + " \"--label\", label,\n", + " \"--min-dist\", str(min_dist),\n", + " \"--replicate-ratio\", str(replicate_ratio),\n", + " \"--batch-ratio\", str(batch_ratio),\n", + " \"--n-neighbors-rep\", str(n_neighbors_rep),\n", + " \"--n-neighbors-batch\", str(n_neighbors_batch),\n", + " \"--n-neighbors-context\", str(n_neighbors_context),\n", + " \"--min-count\", str(min_count),\n", + " \"--quantile\", str(quantile),\n", + " \"--seed\", str(seed),\n", " ]\n", ")\n", "# fmt: on\n", @@ -591,12 +627,10 @@ ] }, { + "metadata": {}, "cell_type": "code", - "execution_count": null, - "metadata": { - "tags": [] - }, "outputs": [], + "execution_count": null, "source": [ "expression_requirement = 3\n", "requirement_adjust = \"regressive\"\n", @@ -609,18 +643,18 @@ "cmd = \" \".join(\n", " [\n", " \"python3\", \"como/merge_xomics.py\",\n", - " \"--merge-distribution\",\n", - " \"--total-rnaseq-config-file\", f\"{trnaseq_config_file}\",\n", - " \"--mrnaseq-config-file\", f\"{mrnaseq_config_file}\",\n", - " # \"--scrnaseq-config-file\", f\"{scrnaseq_config_file}\", # If using single-cell data, uncomment the start of this line\n", - " # \"--proteomics-config-file\", f\"{proteomics_config_file}\",\n", - " \"--expression-requirement\", f\"{expression_requirement}\",\n", - " \"--requirement-adjust\", f\"{requirement_adjust}\",\n", - " \"--total-rnaseq-weight\", f\"{total_rna_weight}\",\n", - " \"--mrnaseq-weight\", f\"{mrna_weight}\",\n", - " # \"--single-cell-rnaseq-weight\", f\"{single_cell_weight}\", # If using single-cell data, uncomment the start of this line\n", - " \"--protein-weight\", f\"{proteomics_weight}\",\n", - " \"--no-hc\",\n", + " \"--merge-zfpkm-distribution\",\n", + " \"--total-rnaseq-config-file\", trnaseq_config_file,\n", + " \"--mrnaseq-config-file\", mrnaseq_config_file,\n", + " # \"--scrnaseq-config-file\", scrnaseq_config_file, # If using single-cell data, uncomment the start of this line\n", + " # \"--proteomics-config-file\", proteomics_config_file, # If using proteomics data, uncomment the start of this line\n", + " \"--requirement-adjust\", requirement_adjust,\n", + " \"--expression-requirement\", str(expression_requirement),\n", + " \"--total-rnaseq-weight\", str(total_rna_weight),\n", + " \"--mrnaseq-weight\", str(mrna_weight),\n", + " \"--single-cell-rnaseq-weight\", str(single_cell_weight),\n", + " \"--protein-weight\", str(proteomics_weight),\n", + " \"--no-high-confidence\",\n", " ]\n", ")\n", "# fmt: on\n", @@ -696,9 +730,124 @@ }, { "cell_type": "code", - "execution_count": null, + "execution_count": 31, "metadata": {}, - "outputs": [], + "outputs": [ + { + "name": "stdout", + "output_type": "stream", + "text": [ + "/Users/joshl/Projects/COMO/.venv/lib/python3.10/site-packages/cobamp/wrappers/external_wrappers.py:9: UserWarning: \n", + "The wrappers.external_wrappers module will be deprecated in a future release in favour of the wrappers module. \n", + " Available ModelObjectReader classes can still be loaded using cobamp.wrappers.. An appropriate model \n", + " reader can also be created using the get_model_reader function on cobamp.wrappers\n", + " warnings.warn(\n", + "\u001B[32m2024-12-04 10:27:00.656\u001B[0m | \u001B[1mINFO \u001B[0m | \u001B[36m__main__\u001B[0m:\u001B[36mcreate_context_specific_model\u001B[0m:\u001B[36m654\u001B[0m - \u001B[1mCreating 'naiveB' model using 'IMAT' reconstruction and 'GUROBI' solver\u001B[0m\n", + "\u001B[32m2024-12-04 10:27:00.656\u001B[0m | \u001B[33m\u001B[1mWARNING \u001B[0m | \u001B[36mcomo.project\u001B[0m:\u001B[36m__init__\u001B[0m:\u001B[36m30\u001B[0m - \u001B[33m\u001B[1m'data_dir' not provided to Config, using /Users/joshl/Projects/COMO/main/data\u001B[0m\n", + "\u001B[32m2024-12-04 10:27:00.656\u001B[0m | \u001B[33m\u001B[1mWARNING \u001B[0m | \u001B[36mcomo.project\u001B[0m:\u001B[36m__init__\u001B[0m:\u001B[36m36\u001B[0m - \u001B[33m\u001B[1m'config_dir' not provided to Config, using /Users/joshl/Projects/COMO/main/data/config_sheets\u001B[0m\n", + "\u001B[32m2024-12-04 10:27:00.656\u001B[0m | \u001B[33m\u001B[1mWARNING \u001B[0m | \u001B[36mcomo.project\u001B[0m:\u001B[36m__init__\u001B[0m:\u001B[36m42\u001B[0m - \u001B[33m\u001B[1m'results_dir' not provided to Config, using /Users/joshl/Projects/COMO/main/data/results\u001B[0m\n", + "Set parameter WLSAccessID\n", + "Set parameter WLSSecret\n", + "Set parameter LicenseID to value 898845\n", + "Academic license 898845 - for non-commercial use only - registered to jl___@huskers.unl.edu\n", + "No defined compartments in model GeneralModelUpdatedV2. Compartments will be deduced heuristically using regular expressions.\n", + "Using regular expression found the following compartments:c, e, g, i, l, m, n, r, x\n", + "/Users/joshl/Projects/COMO/main/como/utils.py:154: FutureWarning: Setting an item of incompatible dtype is deprecated and will raise in a future error of pandas. Value '['1' '10' '100' ... '9992' '9993' '9997']' has dtype incompatible with int64, please explicitly cast to a compatible dtype first.\n", + " expression_data.loc[:, \"entrez_gene_id\"] = expression_data[\"entrez_gene_id\"].astype(str)\n", + "\u001B[32m2024-12-04 10:27:04.957\u001B[0m | \u001B[1mINFO \u001B[0m | \u001B[36m__main__\u001B[0m:\u001B[36m_map_expression_to_reaction\u001B[0m:\u001B[36m391\u001B[0m - \u001B[1mMapped gene expression to reactions, found 2806 error(s).\u001B[0m\n", + "\u001B[32m2024-12-04 10:27:04.959\u001B[0m | \u001B[33m\u001B[1mWARNING \u001B[0m | \u001B[36m__main__\u001B[0m:\u001B[36m_build_model\u001B[0m:\u001B[36m468\u001B[0m - \u001B[33m\u001B[1mThe force reaction 'ICDH_m' was not found in the general reference_model. Check BiGG, or the relevant database for your general reference_model, for synonyms.\u001B[0m\n", + "\u001B[32m2024-12-04 10:27:04.959\u001B[0m | \u001B[33m\u001B[1mWARNING \u001B[0m | \u001B[36m__main__\u001B[0m:\u001B[36m_build_model\u001B[0m:\u001B[36m468\u001B[0m - \u001B[33m\u001B[1mThe force reaction 'CATC140_c' was not found in the general reference_model. Check BiGG, or the relevant database for your general reference_model, for synonyms.\u001B[0m\n", + "Read LP format model from file /var/folders/6s/9l9z74v90tn8lprrp7fchb9w0000gn/T/tmprdfbwfb2.lp\n", + "Reading time = 0.02 seconds\n", + ": 5837 rows, 21220 columns, 81018 nonzeros\n", + "/Users/joshl/Projects/COMO/.venv/lib/python3.10/site-packages/cobra/core/group.py:147: UserWarning: need to pass in a list\n", + " warn(\"need to pass in a list\")\n", + "Error encountered trying to >.\n", + "LibSBML error code -3: The requested action could not be performed. This can occur in a variety of contexts, such as passing a null object as a parameter in a situation where it does not make sense to permit a null object.\n", + "Error encountered trying to >.\n", + "LibSBML error code -3: The requested action could not be performed. This can occur in a variety of contexts, such as passing a null object as a parameter in a situation where it does not make sense to permit a null object.\n", + "Error encountered trying to >.\n", + "LibSBML error code -3: The requested action could not be performed. This can occur in a variety of contexts, such as passing a null object as a parameter in a situation where it does not make sense to permit a null object.\n", + "Error encountered trying to >.\n", + "LibSBML error code -3: The requested action could not be performed. This can occur in a variety of contexts, such as passing a null object as a parameter in a situation where it does not make sense to permit a null object.\n", + "Error encountered trying to >.\n", + "LibSBML error code -3: The requested action could not be performed. This can occur in a variety of contexts, such as passing a null object as a parameter in a situation where it does not make sense to permit a null object.\n", + "Error encountered trying to >.\n", + "LibSBML error code -3: The requested action could not be performed. This can occur in a variety of contexts, such as passing a null object as a parameter in a situation where it does not make sense to permit a null object.\n", + "Error encountered trying to >.\n", + "LibSBML error code -3: The requested action could not be performed. This can occur in a variety of contexts, such as passing a null object as a parameter in a situation where it does not make sense to permit a null object.\n", + "Error encountered trying to >.\n", + "LibSBML error code -3: The requested action could not be performed. This can occur in a variety of contexts, such as passing a null object as a parameter in a situation where it does not make sense to permit a null object.\n", + "Error encountered trying to >.\n", + "LibSBML error code -3: The requested action could not be performed. This can occur in a variety of contexts, such as passing a null object as a parameter in a situation where it does not make sense to permit a null object.\n", + "Error encountered trying to >.\n", + "LibSBML error code -3: The requested action could not be performed. This can occur in a variety of contexts, such as passing a null object as a parameter in a situation where it does not make sense to permit a null object.\n", + "Error encountered trying to >.\n", + "LibSBML error code -3: The requested action could not be performed. This can occur in a variety of contexts, such as passing a null object as a parameter in a situation where it does not make sense to permit a null object.\n", + "Error encountered trying to >.\n", + "LibSBML error code -3: The requested action could not be performed. This can occur in a variety of contexts, such as passing a null object as a parameter in a situation where it does not make sense to permit a null object.\n", + "Error encountered trying to >.\n", + "LibSBML error code -3: The requested action could not be performed. This can occur in a variety of contexts, such as passing a null object as a parameter in a situation where it does not make sense to permit a null object.\n", + "Error encountered trying to >.\n", + "LibSBML error code -3: The requested action could not be performed. This can occur in a variety of contexts, such as passing a null object as a parameter in a situation where it does not make sense to permit a null object.\n", + "Error encountered trying to >.\n", + "LibSBML error code -3: The requested action could not be performed. This can occur in a variety of contexts, such as passing a null object as a parameter in a situation where it does not make sense to permit a null object.\n", + "Error encountered trying to >.\n", + "LibSBML error code -3: The requested action could not be performed. This can occur in a variety of contexts, such as passing a null object as a parameter in a situation where it does not make sense to permit a null object.\n", + "Error encountered trying to >.\n", + "LibSBML error code -3: The requested action could not be performed. This can occur in a variety of contexts, such as passing a null object as a parameter in a situation where it does not make sense to permit a null object.\n", + "Error encountered trying to >.\n", + "LibSBML error code -3: The requested action could not be performed. This can occur in a variety of contexts, such as passing a null object as a parameter in a situation where it does not make sense to permit a null object.\n", + "Error encountered trying to >.\n", + "LibSBML error code -3: The requested action could not be performed. This can occur in a variety of contexts, such as passing a null object as a parameter in a situation where it does not make sense to permit a null object.\n", + "Error encountered trying to >.\n", + "LibSBML error code -3: The requested action could not be performed. This can occur in a variety of contexts, such as passing a null object as a parameter in a situation where it does not make sense to permit a null object.\n", + "Error encountered trying to >.\n", + "LibSBML error code -3: The requested action could not be performed. This can occur in a variety of contexts, such as passing a null object as a parameter in a situation where it does not make sense to permit a null object.\n", + "Error encountered trying to >.\n", + "LibSBML error code -3: The requested action could not be performed. This can occur in a variety of contexts, such as passing a null object as a parameter in a situation where it does not make sense to permit a null object.\n", + "Error encountered trying to >.\n", + "LibSBML error code -3: The requested action could not be performed. This can occur in a variety of contexts, such as passing a null object as a parameter in a situation where it does not make sense to permit a null object.\n", + "Error encountered trying to >.\n", + "LibSBML error code -3: The requested action could not be performed. This can occur in a variety of contexts, such as passing a null object as a parameter in a situation where it does not make sense to permit a null object.\n", + "Error encountered trying to >.\n", + "LibSBML error code -3: The requested action could not be performed. This can occur in a variety of contexts, such as passing a null object as a parameter in a situation where it does not make sense to permit a null object.\n", + "Error encountered trying to >.\n", + "LibSBML error code -3: The requested action could not be performed. This can occur in a variety of contexts, such as passing a null object as a parameter in a situation where it does not make sense to permit a null object.\n", + "Error encountered trying to >.\n", + "LibSBML error code -3: The requested action could not be performed. This can occur in a variety of contexts, such as passing a null object as a parameter in a situation where it does not make sense to permit a null object.\n", + "Error encountered trying to >.\n", + "LibSBML error code -3: The requested action could not be performed. This can occur in a variety of contexts, such as passing a null object as a parameter in a situation where it does not make sense to permit a null object.\n", + "Error encountered trying to >.\n", + "LibSBML error code -3: The requested action could not be performed. This can occur in a variety of contexts, such as passing a null object as a parameter in a situation where it does not make sense to permit a null object.\n", + "Error encountered trying to >.\n", + "LibSBML error code -3: The requested action could not be performed. This can occur in a variety of contexts, such as passing a null object as a parameter in a situation where it does not make sense to permit a null object.\n", + "Error encountered trying to >.\n", + "LibSBML error code -3: The requested action could not be performed. This can occur in a variety of contexts, such as passing a null object as a parameter in a situation where it does not make sense to permit a null object.\n", + "Error encountered trying to >.\n", + "LibSBML error code -3: The requested action could not be performed. This can occur in a variety of contexts, such as passing a null object as a parameter in a situation where it does not make sense to permit a null object.\n", + "Error encountered trying to >.\n", + "LibSBML error code -3: The requested action could not be performed. This can occur in a variety of contexts, such as passing a null object as a parameter in a situation where it does not make sense to permit a null object.\n", + "Error encountered trying to >.\n", + "LibSBML error code -3: The requested action could not be performed. This can occur in a variety of contexts, such as passing a null object as a parameter in a situation where it does not make sense to permit a null object.\n", + "Error encountered trying to >.\n", + "LibSBML error code -3: The requested action could not be performed. This can occur in a variety of contexts, such as passing a null object as a parameter in a situation where it does not make sense to permit a null object.\n", + "Error encountered trying to >.\n", + "LibSBML error code -3: The requested action could not be performed. This can occur in a variety of contexts, such as passing a null object as a parameter in a situation where it does not make sense to permit a null object.\n", + "Error encountered trying to >.\n", + "LibSBML error code -3: The requested action could not be performed. This can occur in a variety of contexts, such as passing a null object as a parameter in a situation where it does not make sense to permit a null object.\n", + "Error encountered trying to >.\n", + "LibSBML error code -3: The requested action could not be performed. This can occur in a variety of contexts, such as passing a null object as a parameter in a situation where it does not make sense to permit a null object.\n", + "Error encountered trying to >.\n", + "LibSBML error code -3: The requested action could not be performed. This can occur in a variety of contexts, such as passing a null object as a parameter in a situation where it does not make sense to permit a null object.\n", + "\u001B[32m2024-12-04 10:27:45.905\u001B[0m | \u001B[32m\u001B[1mSUCCESS \u001B[0m | \u001B[36m__main__\u001B[0m:\u001B[36mcreate_context_specific_model\u001B[0m:\u001B[36m690\u001B[0m - \u001B[32m\u001B[1mSaved output file to /Users/joshl/Projects/COMO/main/data/results/naiveB\u001B[0m\n", + "\u001B[32m2024-12-04 10:27:45.905\u001B[0m | \u001B[1mINFO \u001B[0m | \u001B[36m__main__\u001B[0m:\u001B[36mcreate_context_specific_model\u001B[0m:\u001B[36m691\u001B[0m - \u001B[1mNumber of Genes: 1,477\u001B[0m\n", + "\u001B[32m2024-12-04 10:27:45.905\u001B[0m | \u001B[1mINFO \u001B[0m | \u001B[36m__main__\u001B[0m:\u001B[36mcreate_context_specific_model\u001B[0m:\u001B[36m692\u001B[0m - \u001B[1mNumber of Metabolites: 3,204\u001B[0m\n", + "\u001B[32m2024-12-04 10:27:45.905\u001B[0m | \u001B[1mINFO \u001B[0m | \u001B[36m__main__\u001B[0m:\u001B[36mcreate_context_specific_model\u001B[0m:\u001B[36m693\u001B[0m - \u001B[1mNumber of Reactions: 5,263\u001B[0m\n", + "Warning: environment still referenced so free is deferred (Continue to use WLS)\n" + ] + } + ], "source": [ "# Set your objectives before running!\n", "objective_dict = {\"naiveB\": \"biomass_maintenance\", \"smB\": \"biomass_maintenance\"}\n", @@ -744,18 +893,18 @@ " cmd = \" \".join(\n", " [\n", " \"python3\", \"como/create_context_specific_model.py\",\n", - " \"--context\", f\"{context}\",\n", - " \"--reference-model-filepath\", f\"{general_model_filepath}\",\n", - " \"--active-genes-filepath\", f\"{active_genes_filepath}\",\n", - " \"--objective\", f\"{objective}\",\n", - " \"--boundary-reactions-filepath\", f\"{boundary_reactions_filepath}\",\n", - " # \"--exclude-reactions-filepath\", f\"{exclude_reactions_filepath}\",\n", - " \"--force-reactions-filepath\", f\"{force_reactions_filepath}\",\n", - " \"--algorithm\", f\"{recon_algorithm}\",\n", - " \"--low-threshold\", f\"{low_threshold}\",\n", - " \"--high-threshold\", f\"{high_threshold}\",\n", - " \"--solver\", f\"{solver}\",\n", - " \"--output-filetypes\", f\"{output_filetypes}\",\n", + " \"--context\", context,\n", + " \"--reference-model-filepath\", general_model_filepath,\n", + " \"--active-genes-filepath\", active_genes_filepath,\n", + " \"--objective\", objective,\n", + " \"--boundary-reactions-filepath\", boundary_reactions_filepath,\n", + " # \"--exclude-reactions-filepath\", exclude_reactions_filepath,\n", + " \"--force-reactions-filepath\", force_reactions_filepath,\n", + " \"--algorithm\", recon_algorithm,\n", + " \"--low-threshold\", str(low_threshold),\n", + " \"--high-threshold\", str(high_threshold),\n", + " \"--solver\", solver,\n", + " \"--output-filetypes\", output_filetypes,\n", " ]\n", " )\n", " # fmt: on\n", @@ -884,16 +1033,14 @@ { "cell_type": "code", "execution_count": null, - "metadata": { - "tags": [] - }, + "metadata": {}, "outputs": [], "source": [ "disease_names = [\"arthritis\", \"lupus_a\", \"lupus_b\"]\n", "data_source = \"rnaseq\"\n", "taxon_id = \"human\"\n", "\n", - "from como.como_utilities import stringlist_to_list\n", + "from como.utils import stringlist_to_list\n", "\n", "for context_name in stringlist_to_list(context_names):\n", " disease_config_file = f\"disease_data_inputs_{context_name}.xlsx\"\n", @@ -902,10 +1049,10 @@ " cmd = \" \".join(\n", " [\n", " \"python3\", \"como/disease_analysis.py\",\n", - " \"--context-name\", f\"{context_name}\",\n", - " \"--config-file\", f\"{disease_config_file}\",\n", - " \"--data-source\", f\"{data_source}\",\n", - " \"--taxon-id\", f\"{taxon_id}\",\n", + " \"--context-name\", context_name,\n", + " \"--config-file\", disease_config_file,\n", + " \"--data-source\", data_source,\n", + " \"--taxon-id\", str(taxon_id),\n", " ]\n", " )\n", " # fmt: on\n", @@ -950,9 +1097,7 @@ { "cell_type": "code", "execution_count": null, - "metadata": { - "tags": [] - }, + "metadata": {}, "outputs": [], "source": [ "# Knock out simulation for the analyzed tissues and diseases\n", @@ -965,7 +1110,7 @@ "import json\n", "import os\n", "\n", - "from como.como_utilities import stringlist_to_list\n", + "from como.utils import stringlist_to_list\n", "from como.project import Config\n", "\n", "config = Config()\n", @@ -992,20 +1137,20 @@ " up_regulated_disease_genes = disease_files[\"up_regulated\"]\n", "\n", " if context in model_files.keys():\n", - " tissueSpecificModelfile = model_files[context]\n", + " tissue_specific_model_filepath = model_files[context]\n", " else:\n", - " tissueSpecificModelfile = os.path.join(config.data_dir, \"results\", context, f\"{context}_SpecificModel_{recon_algorithm}.mat\")\n", + " tissue_specific_model_filepath = os.path.join(config.data_dir, \"results\", context, f\"{context}_SpecificModel_{recon_algorithm}.mat\")\n", "\n", " # fmt: off\n", " cmd = [\n", " \"python3\", \"como/knock_out_simulation.py\",\n", - " \"--context-model\", f\"{tissueSpecificModelfile}\",\n", - " \"--context-name\", f\"{context}\",\n", - " \"--disease-name\", f\"{disease}\",\n", - " \"--disease-up\", f\"{up_regulated_disease_genes}\",\n", - " \"--disease-down\", f\"{down_regulated_disease_genes}\",\n", - " \"--raw-drug-file\", f\"{drug_raw_file}\",\n", - " \"--solver\", f\"{sovler}\",\n", + " \"--context-model\", tissue_specific_model_filepath,\n", + " \"--context-name\", context,\n", + " \"--disease-name\", disease,\n", + " \"--disease-up\", up_regulated_disease_genes,\n", + " \"--disease-down\", down_regulated_disease_genes,\n", + " \"--raw-drug-file\", drug_raw_file,\n", + " \"--solver\", sovler,\n", " # \"--test-all\"\n", " ]\n", " # fmt: on\n", @@ -1034,7 +1179,7 @@ "name": "python", "nbconvert_exporter": "python", "pygments_lexer": "ipython3", - "version": "3.11.10" + "version": "3.10.15" }, "toc-autonumbering": true, "toc-showcode": true diff --git a/main/como/__init__.py b/main/como/__init__.py index 8cb36e7a..05100d09 100644 --- a/main/como/__init__.py +++ b/main/como/__init__.py @@ -1 +1,11 @@ -from .como_utilities import stringlist_to_list \ No newline at end of file +import pandas as pd + +from como.project import Config +from como.utils import stringlist_to_list + +__all__ = ["stringlist_to_list", "Config"] +__version__ = "1.10.0" + + +def return_placeholder_data() -> pd.DataFrame: + return pd.DataFrame(data=0, index=pd.Index(data=[0], name="entrez_gene_id"), columns=["expressed", "top"]) diff --git a/main/como/cluster_rnaseq.py b/main/como/cluster_rnaseq.py index 9cc3ae0c..dba5499c 100644 --- a/main/como/cluster_rnaseq.py +++ b/main/como/cluster_rnaseq.py @@ -1,21 +1,79 @@ +from __future__ import annotations + import argparse -from pathlib import Path +from curses.ascii import isdigit +from dataclasses import dataclass +from typing import Any import numpy as np -import rpy2_api -from como_utilities import stringlist_to_list -from project import Config -# read and translate R functions -configs = Config() -r_file_path = Path(configs.code_dir, "rscripts", "cluster_samples.R") +from como.utils import stringlist_to_list + + +@dataclass +class _Arguments: + context_names: list[str] + filtering_technique: str + cluster_algorithm: str + label_plot: str + min_distance: float + replicate_ratio: Any + batch_ratio: Any + num_replicate_neighbors: Any + num_batch_neighbors: Any + num_context_neighbors: Any + min_active_count: int | str + quantile: Any + seed: int + + def __post_init__(self): # noqa: C901, ignore too complex + self.filtering_technique = self.filtering_technique.lower() + self.cluster_algorithm = self.cluster_algorithm.lower() + + if self.seed == -1: + self.seed = np.random.randint(0, 100_000) + + if (isdigit(self.min_active_count) and int(self.min_active_count) < 0) or self.min_active_count != "default": + raise ValueError("--min-count must be either 'default' or an integer > 0") + + if (isdigit(self.quantile) and 0 > int(self.quantile) > 100) or self.quantile != "default": + raise ValueError("--quantile must be either 'default' or an integer between 0 and 100") + + if (isdigit(self.replicate_ratio) and 0 > self.replicate_ratio > 1.0) or self.replicate_ratio != "default": + raise ValueError("--rep-ratio must be either 'default' or a float between 0 and 1") + + if (isdigit(self.batch_ratio) and 0 > self.batch_ratio > 1.0) or self.batch_ratio != "default": + raise ValueError("--batch-ratio must be either 'default' or a float between 0 and 1") + + if self.filtering_technique.lower() not in {"quantile", "tpm", "cpm", "zfpkm"}: + raise ValueError("--technique must be either 'quantile', 'tpm', 'cpm', 'zfpkm'") + + if self.filtering_technique.lower() == "tpm": + self.filtering_technique = "quantile" + if self.cluster_algorithm.lower() not in {"mca", "umap"}: + raise ValueError("--clust_algo must be either 'mca', 'umap'") -def main() -> None: - """ - Cluster RNA-seq Data - """ + if 0 > self.min_distance > 1.0: + raise ValueError("--min_dist must be a float between 0 and 1") + if ( + isdigit(self.num_replicate_neighbors) and self.num_replicate_neighbors < 1 + ) or self.num_replicate_neighbors != "default": + raise ValueError("--n-neighbors-rep must be either 'default' or an integer > 1") + + if ( + isdigit(self.num_batch_neighbors) and self.num_batch_neighbors < 1 + ) or self.num_batch_neighbors != "default": + raise ValueError("--n-neighbors-batch must be either 'default' or an integer > 1") + + if ( + isdigit(self.num_context_neighbors) and self.num_context_neighbors < 1 + ) or self.num_context_neighbors != "default": + raise ValueError("--n-neighbors-context must be either 'default' or an integer > 1") + + +def _parse_args() -> _Arguments: parser = argparse.ArgumentParser( prog="cluster_rnaseq.py", description="Cluster RNA-seq Data using Multiple Correspondence Analysis or UMAP. Clusters at the replicate, " @@ -24,49 +82,42 @@ def main() -> None: "https://github.com/HelikarLab/MADRID or email babessell@gmail.com", ) parser.add_argument( - "-n", "--context-names", type=str, required=True, dest="context_names", - help="""Tissue/cell name of models to generate. If making multiple models in a batch, then - use the format: \"['context1', 'context2', ... etc]\". Note the outer double-quotes and the - inner single-quotes are required to be interpreted. This a string, not a python list""", + help="Tissue/cell name of models to generate", ) parser.add_argument( - "-t", - "--filt-technique", + "--filtering-technique", type=str, required=True, - dest="technique", + dest="filtering_technique", help="'zfpkm', 'quantile', or 'cpm'", ) parser.add_argument( - "-a", "--cluster-algorithm", type=str, required=False, default="umap", - dest="clust_algo", - help="""Clustering algorithm to use. 'mca' or 'umap'.""", + dest="cluster_algorithm", + help="Clustering algorithm to use. 'mca' or 'umap'.", ) parser.add_argument( - "-l", - "--label", + "--label-plot", type=str, required=False, default=True, - dest="label", - help="""True to label replicate/batch/context names on the plots. May be ugly for large sets""", + dest="label_plot", + help="Set to True to label replicate/batch/context names on the plots. May be ugly for large sets", ) parser.add_argument( - "-d", - "--min-dist", + "--min-distance", type=float, required=False, default=0.01, - dest="min_dist", - help="""Minimum distance for UMAP clustering. Must be between 0 and 1""", + dest="min_distance", + help="Minimum distance for UMAP clustering. Must be between 0 and 1", ) parser.add_argument( "-r", @@ -74,8 +125,8 @@ def main() -> None: type=str, required=False, default=0.9, - dest="rep_ratio", - help="""Ratio of genes active in replicates for a batch/study to be active""", + dest="replicate_ratio", + help="Ratio of genes active in replicates for a batch/study to be active", ) parser.add_argument( "-b", @@ -84,188 +135,57 @@ def main() -> None: required=False, default=0.9, dest="batch_ratio", - help="""Ratio of genes active in a batch/study to be active in the context""", + help="Ratio of genes active in a batch/study to be active in the context", ) parser.add_argument( - "-nr", - "--n-neighbors-rep", + "--num-replicate-neighbors", type=str or float, required=False, default="default", - dest="n_neigh_rep", - help="""N nearest neighbors for replicate clustering, 'default' is total number of replicates""", + dest="num_replicate_neighbors", + help="N nearest neighbors for replicate clustering, 'default' is total number of replicates", ) parser.add_argument( "-nb", - "--n-neighbors-batch", + "--num-batch-neighbors", type=str or float, required=False, default="default", - dest="n_neigh_batch", - help="""N nearest neighbors for batch clustering, 'default' is total number of batches""", + dest="num_batch_neighbors", + help="N nearest neighbors for batch clustering, 'default' is total number of batches", ) parser.add_argument( - "-nc", - "--n-neighbors-context", + "--num-context-neighbors", type=str or float, required=False, default="default", - dest="n_neigh_cont", - help="""N nearest neighbors for context clustering, 'default' is total number of contexts""", + dest="num_context_neighbors", + help="N nearest neighbors for context clustering, 'default' is total number of contexts", ) parser.add_argument( - "-c", - "--min-count", + "--min-active-count", type=str or int, required=False, default="default", - dest="min_count", - help="""Ratio of active genes in a batch/study to be active in the context""", + dest="min_active_count", + help="Ratio of active genes in a batch/study to be active in the context", ) parser.add_argument( - "-q", "--quantile", type=str or int, required=False, default=0.5, dest="quantile", - help="""Ratio of active genes in a batch/study to be active in the context""", + help="Ratio of active genes in a batch/study to be active in the context", ) parser.add_argument( - "-s", "--seed", type=int, required=False, default=-1, dest="seed", - help="""Random seed for clustering algorithm initialization""", + help="Random seed for clustering algorithm initialization", ) args = parser.parse_args() - - context_names = stringlist_to_list(args.context_names) - technique = args.technique.lower() - clust_algo = args.clust_algo.lower() - label = args.label - rep_ratio = args.rep_ratio - batch_ratio = args.batch_ratio - min_count = args.min_count - quantile = args.quantile - min_dist = args.min_dist - n_neigh_rep = args.n_neigh_rep - n_neigh_batch = args.n_neigh_batch - n_neigh_cont = args.n_neigh_cont - - # Set a random seed if none provided - if int(args.seed) == -1: - seed = np.random.randint(0, 100000) - else: - seed = args.seed - - if isinstance(min_count, str) and min_count.lower() == "default": - try: - min_count = int(min_count) - except ValueError: - raise ValueError("--min-count must be either 'default' or an integer > 0") - if not isinstance(min_count, str) and min_count < 0: - raise ValueError("--min-count must be either 'default' or an integer > 0") - - if isinstance(quantile, str) and not quantile.lower() == "default": - try: - quantile = int(quantile) - except ValueError: - raise ValueError("--quantile must be either 'default' or an integer between 0 and 100") - if not isinstance(quantile, str) and 0 > quantile > 100: - raise ValueError("--quantile must be either 'default' or an integer between 0 and 100") - - if isinstance(rep_ratio, str) and not rep_ratio.lower() == "default": - try: - rep_ratio = float(rep_ratio) - except ValueError: - raise ValueError("--rep-ratio must be 'default' or a float between 0 and 1") - if not isinstance(rep_ratio, str) and 0 > rep_ratio > 1.0: - raise ValueError("--rep-ratio must be 'default' or a float between 0 and 1") - - if isinstance(batch_ratio, str) and not batch_ratio.lower() == "default": - try: - batch_ratio = float(batch_ratio) - except ValueError: - raise ValueError("--batch-ratio must be 'default' or a float between 0 and 1") - if not isinstance(batch_ratio, str) and 0 > batch_ratio > 1.0: - raise ValueError("--batch-ratio must be 'default' or a float between 0 and 1") - - if technique.lower() not in ["quantile", "tpm", "cpm", "zfpkm"]: - raise ValueError("--technique must be either 'quantile', 'tpm', 'cpm', 'zfpkm'") - - if technique.lower() == "tpm": - technique = "quantile" - - if clust_algo.lower() not in ["mca", "umap"]: - raise ValueError("--clust_algo must be either 'mca', 'umap'") - - if not isinstance(min_dist, str) and 0 > min_dist > 1.0: - raise ValueError("--min_dist must be a float between 0 and 1") - - if isinstance(n_neigh_rep, str) and not n_neigh_rep.lower() == "default": - try: - n_neigh_rep = int(n_neigh_rep) - except ValueError: - raise ValueError( - "--n_neigh_rep must be either 'default' or an integer greater than 1 and less than or equal to " - "the total number of replicates being clustered across all contexts." - ) - if not isinstance(n_neigh_rep, str) and n_neigh_rep < 2: - raise ValueError( - "--n_neigh_rep must be either 'default' or an integer greater than 1 and less than or equal to " - "the total number of replicates being clustered across all contexts." - ) - - if isinstance(n_neigh_batch, str) and not n_neigh_batch.lower() == "default": - try: - n_neigh_batch = int(n_neigh_batch) - except ValueError: - raise ValueError( - "--n_neigh_batch must be either 'default' or an integer greater than 1 and less than or equal to " - "the total number of batches being clustered across all contexts." - ) - if not isinstance(n_neigh_batch, str) and n_neigh_batch < 2: - raise ValueError( - "--n_neigh_batch must be either 'default' or an integer greater than 1 and less than or equal to " - "the total number of batches being clustered across all contexts." - ) - - if isinstance(n_neigh_cont, str) and not n_neigh_cont.lower() == "default": - try: - n_neigh_cont = int(n_neigh_cont) - except ValueError: - raise ValueError( - "--n_neigh_batch must be either 'default' or an integer greater than 1 and less than or equal to " - "the total number of batches being clustered across all contexts." - ) - if not isinstance(n_neigh_cont, str) and n_neigh_cont < 2: - raise ValueError( - "--n_neigh_context must be either 'default' or an integer greater than 1 and less than or equal to " - "the total number of contexts being clustered." - ) - - cluster_samples = rpy2_api.Rpy2( - r_file_path=r_file_path, - wd=configs.result_dir, - context_names=context_names, - technique=technique, - clust_algo=clust_algo, - label=label, - min_dist=min_dist, - n_neigh_rep=n_neigh_rep, - n_neigh_batch=n_neigh_batch, - n_neigh_cont=n_neigh_cont, - rep_ratio=rep_ratio, - batch_ratio=batch_ratio, - quantile=quantile, - min_count=min_count, - seed=seed, - ) - cluster_samples.call_function("cluster_samples_main") - - -if __name__ == "__main__": - main() + args.context_names = stringlist_to_list(args.context_names) + return _Arguments(**vars(args)) diff --git a/main/como/cluster_sources.py b/main/como/cluster_sources.py index 4f40c1c9..c9e91dbd 100644 --- a/main/como/cluster_sources.py +++ b/main/como/cluster_sources.py @@ -1,4 +1,5 @@ from pathlib import Path + import rpy2_api # read and translate R functions @@ -17,5 +18,5 @@ source_type=source_type, use_trna=use_trna, use_mrna=use_mrna, - binarize_data=binarize_data + binarize_data=binarize_data, ).call_function("cluster_sources_main") diff --git a/main/como/combine_distributions.py b/main/como/combine_distributions.py index 2ef2d289..ed69413b 100644 --- a/main/como/combine_distributions.py +++ b/main/como/combine_distributions.py @@ -1,19 +1,463 @@ from pathlib import Path -from typing import Literal, Union +import numpy as np +import pandas as pd +import plotly.express as px +from loguru import logger -def parse_contexts_zfpkm(wd: Union[str, Path], contexts: list[str], prep: Literal["mrna", "total"]): - wd: Path = Path(wd) - batches = [] +def _get_batch_name(x): + return Path(x).stem + + +def _parse_contexts_zfpkm(wd, contexts, prep): + batches = {} + for context in contexts: + files = (wd / context / prep).glob(f"rnaseq_{prep}_{context}.csv") + batches[context] = [_get_batch_name(f) for f in files] + return batches + + +def _parse_contexts_zumi(wd, contexts, prep): + batches = {} for context in contexts: - dir_name = Path(wd, context, prep) - files = dir_name.glob(f"zFPKM_Matrix_{prep}_*.csv") - batches += [Path(file).stem for file in files] + files = (wd / context / prep).glob(f"*{prep}*.csv") + batches[context] = [_get_batch_name(f) for f in files] + return batches + +def _parse_contexts_zscore_prot(wd, contexts): + batches = {} + for context in contexts: + files = (wd / context / "proteomics").glob("protein_zscore_Matrix_*.csv") + batches[context] = [_get_batch_name(f) for f in files] return batches -if __name__ == "__main__": - result = parse_contexts_zfpkm(wd="/Users/joshl/PycharmProjects/COMO/main/data/results", contexts=["naiveB"], prep="total") - print(result) +def _merge_batch(wd, context, batch): + files = list(wd.glob(f"*{batch}*")) + nrep = [] + if not files: + raise ValueError(f"No files found for {context} - {batch}") + + for f in files: + zmat = pd.read_csv(f) + zmat.columns = pd.Index([c.lower() for c in zmat.columns]) + zmat["entrez_gene_id"] = zmat["entrez_gene_id"].astype(str).str.split("//").str[0] + + zmat = zmat.astype({col: float for col in zmat.columns if col != "entrez_gene_id"}) + zmat = zmat.astype({"entrez_gene_id": str}) + zmat = zmat.groupby("entrez_gene_id").max().reset_index() + zmat = zmat.dropna() + + nrep.append(zmat.shape[1] - 1) + entrez_gene = zmat["entrez_gene_id"] + rep_names = zmat.columns + zmat = pd.concat([zmat[col] for col in zmat.columns[1:]], axis=1) + zmat = pd.concat([entrez_gene, zmat], axis=1) + zmat = zmat.dropna() + zmat.columns = rep_names + + stack_df = pd.concat( + [ + pd.DataFrame( + {"entrez_gene_id": zmat["entrez_gene_id"], "zscore": zmat[col].astype(float), "source": col} + ) + for col in zmat.columns[1:] + ] + ) + + plot_name_png = wd / "figures" / f"plot_{context}_{Path(f).stem}.png" + + fig = px.histogram( + stack_df, + x="zscore", + color="source", + nbins=100, # Adjust as needed + marginal="rug", + title=f"Z-score Distribution for {context} - {Path(f).stem}", + ) + + fig.update_layout(xaxis_title="Z-score", yaxis_title="Frequency", font={"family": "sans-serif", "size": 12}) + + # Simplified plot for many sources (optional) + if len(stack_df["source"].unique()) > 10: + fig.update_layout(showlegend=False) + + plot_name_png.parent.mkdir(parents=True, exist_ok=True) + fig.write_image(plot_name_png) + + return zmat, nrep + + +def _combine_batch_zdistro(wd, context, batch, zmat): + plot_name_png = wd / "figures" / f"plot_{context}_{batch}_combine_distro.png" + + def weighted_z(x): + floor_score = -6 + ceil_score = 6 + x = np.array(x, dtype=float) + result = np.sum(x) / np.sqrt(len(x)) + return np.clip(result, floor_score, ceil_score) + + combine_z = zmat + if zmat.shape[1] > 2: + combine_z = np.apply_along_axis(weighted_z, axis=1, arr=zmat.iloc[:, 1:].values) + merge_df = pd.concat([zmat, pd.Series(combine_z, name="combined")], axis=1) + combine_z = pd.DataFrame({"entrez_gene_id": zmat["entrez_gene_id"].astype(str), "combine_z": combine_z}) + + stack_df = pd.concat( + [ + pd.DataFrame( + {"entrez_gene_id": merge_df["entrez_gene_id"], "zscore": merge_df[col].astype(float), "source": col} + ) + for col in merge_df.columns[1:] + ] + ) + + # Simplified plot for many sources (optional) + if len(stack_df["source"].unique()) > 10: + stack_df = stack_df[stack_df["source"] == "combined"] + + fig = px.histogram( + stack_df, + x="zscore", + color="source", + nbins=100, + marginal="rug", + title=f"Combined Z-score Distribution for {context} - {batch}", + ) + fig.update_layout(xaxis_title="Z-score", yaxis_title="Frequency", font={"family": "sans-serif", "size": 12}) + fig.write_image(plot_name_png) + + return combine_z + + +def _combine_context_zdistro(wd, context, n_reps, zmat): + plot_name_png = wd / "figures" / f"plot_{context}_combine_batches_distro.png" + + def weighted_z(x, n_reps): + floor_score = -6 + ceil_score = 6 + x = np.array(x, dtype=float) + nas = np.where(np.isnan(x))[0] + if len(nas) > 0: + x = np.delete(x, nas) + n_reps = np.delete(n_reps, nas) + weights = n_reps / np.sum(n_reps) + numer = np.sum(weights * x) + denom = np.sqrt(np.sum(weights**2)) + result = numer / denom + return np.clip(result, floor_score, ceil_score) + + if zmat.shape[1] > 2: + combine_z = np.apply_along_axis(weighted_z, axis=1, arr=zmat.iloc[:, 1:].values, n_reps=n_reps) + merge_df = pd.concat([zmat, pd.Series(combine_z, name="combined")], axis=1) + combine_z = pd.DataFrame({"entrez_gene_id": zmat["entrez_gene_id"].astype(str), "combine_z": combine_z}) + + stack_df = pd.concat( + [ + pd.DataFrame( + {"entrez_gene_id": merge_df["entrez_gene_id"], "zscore": merge_df[col].astype(float), "source": col} + ) + for col in merge_df.columns[1:] + ] + ) + + fig = px.histogram( + stack_df, + x="zscore", + color="source", + nbins=100, # Adjust as needed + marginal="rug", + title=f"Combined Batches Z-score Distribution for {context}", + ) + + fig.update_layout(xaxis_title="Z-score", yaxis_title="Frequency", font=dict(family="sans-serif", size=12)) # noqa: C408 + + fig.write_image(plot_name_png) + + else: + combine_z = zmat + combine_z.columns = ["entrez_gene_id", "combine_z"] + + return combine_z + + +def _combine_omics_zdistros( + wd, + context, + comb_batches_z_trna, + comb_batches_z_mrna, + comb_batches_z_scrna, + comb_batches_z_prot, + trna_weight: int, + mrna_weight: int, + scrna_weight: int, + proteomics_weight: int, + keep_gene_scores=True, +): + fig_path = wd / context / "figures" + if not fig_path.exists(): + fig_path.mkdir(parents=True) + plot_name_png = fig_path / f"plot_{context}_combine_omics_distro.png" + + weights = [] + names = [] + dfs = [] + counter = 0 + if comb_batches_z_trna is not None: + counter += 1 + weights.append(trna_weight) + names.append("total") + dfs.append(comb_batches_z_trna) + if comb_batches_z_mrna is not None: + counter += 1 + weights.append(mrna_weight) + names.append("polyA") + dfs.append(comb_batches_z_mrna) + if comb_batches_z_scrna is not None: + counter += 1 + weights.append(scrna_weight) + names.append("singleCell") + dfs.append(comb_batches_z_scrna) + if comb_batches_z_prot is not None: + counter += 1 + weights.append(proteomics_weight) + names.append("proteome") + dfs.append(comb_batches_z_prot) + + def weighted_z(x, weights): + floor_score = -6 + ceil_score = 10 + x = np.array(x, dtype=float) + nas = np.where(np.isnan(x))[0] + if len(nas) > 0: + x = np.delete(x, nas) + weights = np.delete(weights, nas) + weights = weights / np.sum(weights) + numer = np.sum(weights * x) + denom = np.sqrt(np.sum(weights**2)) + result = numer / denom + return np.clip(result, floor_score, ceil_score) + + zmat = dfs[0].copy() + zmat.columns = ["entrez_gene_id", names[0]] + for i in range(1, counter): + add_df = dfs[i] + add_df.columns = ["entrez_gene_id", names[i]] + zmat = pd.merge(zmat, add_df, on="entrez_gene_id", how="outer") + + if zmat.shape[1] > 2: + combine_z = np.apply_along_axis(weighted_z, axis=1, arr=zmat.iloc[:, 1:].values, weights=weights) + else: + combine_z = zmat.iloc[:, 1:].values + + combine_z = combine_z.ravel() + merge_df = pd.concat([zmat, pd.Series(combine_z, name="combined")], axis=1) + combine_z = pd.DataFrame({"entrez_gene_id": zmat["entrez_gene_id"].astype(str), "combine_z": combine_z}) + + stack_df = pd.concat( + [ + pd.DataFrame( + {"entrez_gene_id": merge_df["entrez_gene_id"], "zscore": merge_df[col].astype(float), "source": col} + ) + for col in merge_df.columns[1:] + ] + ) + + fig = px.histogram( + stack_df, + x="zscore", + color="source", + nbins=100, # Adjust as needed + marginal="rug", + title=f"Combined Omics Z-score Distribution for {context}", + ) + + fig.update_layout(xaxis_title="Z-score", yaxis_title="Frequency", font={"family": "sans-serif", "size": 12}) + + fig.write_image(plot_name_png) + + return combine_z + + +def _combine_zscores( # noqa: C901 + working_dir, + context_names, + global_use_mrna, + global_use_trna, + global_use_scrna, + global_use_proteins, + keep_gene_scores, + global_trna_weight, + global_mrna_weight, + global_scrna_weight, + global_protein_weight, +): + working_dir = Path(working_dir) + figure_output_dir = working_dir / "figures" + figure_output_dir.mkdir(parents=True, exist_ok=True) + + global_trna_batches = _parse_contexts_zfpkm(working_dir, context_names, "total") + global_mrna_batches = _parse_contexts_zfpkm(working_dir, context_names, "mrna") + global_scrna_batches = _parse_contexts_zumi(working_dir, context_names, "scrna") + global_protein_batches = _parse_contexts_zscore_prot(working_dir, context_names) + + for context in context_names: + context_use_trna = global_use_trna + context_use_mrna = global_use_mrna + context_use_scrna = global_use_scrna + context_use_proteins = global_use_proteins + + context_trna_batch = global_trna_batches.get(context, []) + if len(context_trna_batch) == 0 and global_use_trna: + context_use_trna = False + logger.warning(f"No total RNA-seq zFPKM Matrix files found for {context}. Will not use for this context.") + + context_mrna_batch = global_mrna_batches.get(context, []) + if len(context_mrna_batch) == 0 and global_use_mrna: + context_use_mrna = False + logger.warning(f"No polyA RNA-seq zFPKM Matrix files found for {context}. Will not use for this context.") + + context_scrna_batch = global_scrna_batches.get(context, []) + if len(context_scrna_batch) == 0 and global_use_scrna: + context_use_scrna = False + logger.warning(f"No SC RNA-seq zFPKM Matrix files found for {context}. Will not use for this context.") + + context_protein_batch = global_protein_batches.get(context, []) + if len(context_protein_batch) == 0 and global_use_proteins: + context_use_proteins = False + logger.warning(f"No proteomics z-score Matrix files found for {context}. Will not use for this context.") + + comb_batches_z_trna = None + if context_use_trna: + logger.info("Will merge total RNA-seq distributions") + trna_workdir = working_dir / context / "total" + num_reps = [] + merge_z_data = pd.DataFrame() + + for batch in context_trna_batch: + res = _merge_batch(trna_workdir, context, batch) + z_matrix = res[0] + num_reps.extend(res[1]) + combine_z_matrix = _combine_batch_zdistro(trna_workdir, context, batch, z_matrix) + combine_z_matrix.columns = ["entrez_gene_id", batch] + merge_z_data = ( + combine_z_matrix + if merge_z_data.empty + else pd.merge(merge_z_data, combine_z_matrix, on="entrez_gene_id", how="outer") + ) + + comb_batches_z_trna = _combine_context_zdistro(trna_workdir, context, num_reps, merge_z_data) + filename = trna_workdir / f"combined_zFPKM_{context}.csv" + comb_batches_z_trna.to_csv(filename, index=False) + + if not context_use_proteins and not context_use_mrna and not context_use_scrna: + filename = trna_workdir / f"model_scores_{context}.csv" + comb_batches_z_trna.to_csv(filename, index=False) + + comb_batches_z_mrna = None + if context_use_mrna: + logger.info("Will merge polyA enriched RNA-seq distributions") + mrna_workdir = working_dir / context / "mrna" + num_reps = [] + merge_z_data = pd.DataFrame() + for batch in context_mrna_batch: + res = _merge_batch(mrna_workdir, context, batch) + z_matrix = res[0] + num_reps.extend(res[1]) + combine_z_matrix = _combine_batch_zdistro(mrna_workdir, context, batch, z_matrix) + combine_z_matrix.columns = ["entrez_gene_id", batch] + merge_z_data = ( + combine_z_matrix + if merge_z_data.empty + else pd.merge(merge_z_data, combine_z_matrix, on="entrez_gene_id", how="outer") + ) + + comb_batches_z_mrna = _combine_context_zdistro(mrna_workdir, context, num_reps, merge_z_data) + filename = mrna_workdir / f"combined_zFPKM_{context}.csv" + comb_batches_z_mrna.to_csv(filename, index=False) + + if not context_use_proteins and not context_use_trna and not context_use_scrna: + filename = mrna_workdir / f"model_scores_{context}.csv" + comb_batches_z_mrna.to_csv(filename, index=False) + + comb_batches_z_scrna = None + if context_use_scrna: + logger.info(f"Will merge single-cell RNA-seq distributions for {context}") + scrna_workdir = working_dir / context / "scrna" + num_reps = [] + merge_z_data = pd.DataFrame() + for batch in context_scrna_batch: + res = _merge_batch(scrna_workdir, context, batch) + z_matrix = res[0] + num_reps.extend(res[1]) + combine_z_matrix = _combine_batch_zdistro(scrna_workdir, context, batch, z_matrix) + combine_z_matrix.columns = ["entrez_gene_id", batch] + merge_z_data = ( + combine_z_matrix + if merge_z_data.empty + else pd.merge(merge_z_data, combine_z_matrix, on="entrez_gene_id", how="outer") + ) + + comb_batches_z_scrna = _combine_context_zdistro(scrna_workdir, context, num_reps, merge_z_data) + filename = scrna_workdir / f"combined_zFPKM_{context}.csv" + comb_batches_z_scrna.to_csv(filename, index=False) + + if not context_use_proteins and not context_use_trna and not context_use_mrna: + filename = scrna_workdir / f"model_scores_{context}.csv" + comb_batches_z_scrna.to_csv(filename, index=False) + + comb_batches_z_prot = None + if context_use_proteins: + logger.info("Will merge protein abundance distributions") + protein_workdir = working_dir / context / "proteomics" + num_reps = [] + merge_z_data = pd.DataFrame() + for batch in context_protein_batch: + res = _merge_batch(protein_workdir, context, batch) + z_matrix = res[0] + num_reps.extend(res[1]) + combine_z_matrix = _combine_batch_zdistro(protein_workdir, context, batch, z_matrix) + combine_z_matrix.columns = ["entrez_gene_id", batch] + merge_z_data = ( + combine_z_matrix + if merge_z_data.empty + else pd.merge(merge_z_data, combine_z_matrix, on="entrez_gene_id", how="outer") + ) + + comb_batches_z_prot = _combine_context_zdistro(protein_workdir, context, num_reps, merge_z_data) + filename = protein_workdir / f"combined_zscore_proteinAbundance_{context}.csv" + comb_batches_z_prot.to_csv(filename, index=False) + + if not context_use_mrna and not context_use_trna and not context_use_scrna: + filename = protein_workdir / f"model_scores_{context}.csv" + comb_batches_z_prot.to_csv(filename, index=False) + + if ( + comb_batches_z_mrna is None + and comb_batches_z_trna is None + and comb_batches_z_scrna is None + and comb_batches_z_prot is None + ): + logger.critical( + f"The context '{context}' was found in the configuration file but no data was found on disk!" + ) + continue + + comb_omics_z = _combine_omics_zdistros( + wd=working_dir, + context=context, + comb_batches_z_trna=comb_batches_z_trna, + comb_batches_z_mrna=comb_batches_z_mrna, + comb_batches_z_scrna=comb_batches_z_scrna, + comb_batches_z_prot=comb_batches_z_prot, + trna_weight=global_trna_weight, + mrna_weight=global_mrna_weight, + scrna_weight=global_scrna_weight, + proteomics_weight=global_protein_weight, + ) + + filename = working_dir / context / f"model_scores_{context}.csv" + comb_omics_z.to_csv(filename, index=False) diff --git a/main/como/como_utilities.py b/main/como/como_utilities.py deleted file mode 100644 index f1062b7f..00000000 --- a/main/como/como_utilities.py +++ /dev/null @@ -1,141 +0,0 @@ -import io -import sys -import contextlib -import pandas as pd -from typing import Iterator - - -class Compartments: - """ - This enum will be used to convert from compartment "long-hand" to "short-hand" - - Shorthand from: https://cobrapy.readthedocs.io/en/latest/_modules/cobra/medium/annotations.html - - "Extracellular" -> "e" - "golgi" -> "g" - """ - - SHORTHAND = { - "ce": ["cell envelope"], - "c": [ - "cytoplasm", - "cytosol", - "default", - "in", - "intra cellular", - "intracellular", - "intracellular region", - "intracellular space", - ], - "er": ["endoplasmic reticulum"], - "erm": ["endoplasmic reticulum membrane"], - "e": [ - "extracellular", - "extraorganism", - "out", - "extracellular space", - "extra organism", - "extra cellular", - "extra-organism", - "external", - "external medium", - ], - "f": ["flagellum", "bacterial-type flagellum"], - "g": ["golgi", "golgi apparatus"], - "gm": ["golgi membrane"], - "h": ["chloroplast"], - "l": ["lysosome"], - "im": ["mitochondrial intermembrane space"], - "mm": ["mitochondrial membrane"], - "m": ["mitochondrion", "mitochondria"], - "n": ["nucleus"], - "p": ["periplasm", "periplasmic space"], - "x": ["peroxisome", "glyoxysome"], - "u": ["thylakoid"], - "vm": ["vacuolar membrane"], - "v": ["vacuole"], - "w": ["cell wall"], - "s": ["eyespot", "eyespot apparatus", "stigma"], - } - - _REVERSE_LOOKUP = {value.lower(): key for key, values in SHORTHAND.items() for value in values} - - @classmethod - def get(cls, longhand: str) -> str: - return cls._REVERSE_LOOKUP.get(longhand.lower(), None) - - -def stringlist_to_list(stringlist: str | list[str]) -> list[str]: - """ - We are attempting to move to a new method of gathering a list of items from the command line - In doing so, we must deprecate the use of the current method - - If '[' and ']' are present in the first and last items of the list, assume we are using the "old" method of providing context names - - :param stringlist: The "string list" gathered from the command line. Example input: "['mat', 'xml', 'json']" - """ - if isinstance(stringlist, str): - - if stringlist.startswith("[") and stringlist.endswith("]"): - # Remove any brackets from the first and last items; replace quotation marks and commas with nothing - new_list: list[str] = stringlist.strip("[]").replace("'", "").replace(" ", "").split(",") - - # Show a warning if more than one item is present in the list (this means we are using the old method) - print( - "DeprecationWarning: Please use the new method of providing context names, i.e. --output-filetypes 'type1 type2 type3'.") - print( - "If you are using COMO, this can be done by setting the 'context_names' variable to a simple string separated by spaces. Here are a few examples!") - print("context_names = 'cellType1 cellType2 cellType3'") - print("output_filetypes = 'output1 output2 output3'") - print( - "\nYour current method of passing context names will be removed in the future. Please update your variables above accordingly!\n\n") - - else: - new_list: list[str] = stringlist.split(" ") - - return new_list - return stringlist - - -def split_gene_expression_data(expression_data, recon_algorithm="GIMME"): - """ - Splits genes that have mapped to multiple Entrez IDs are formated as "gene12//gene2//gene3" - """ - if recon_algorithm in ["IMAT", "TINIT"]: - expression_data.rename(columns={"ENTREZ_GENE_ID": "Gene", "combine_z": "Data"}, inplace=True) - else: - expression_data.rename(columns={"ENTREZ_GENE_ID": "Gene", "Active": "Data"}, inplace=True) - - expression_data = expression_data.loc[:, ["Gene", "Data"]] - expression_data["Gene"] = expression_data["Gene"].astype(str) - single_gene_names = expression_data[~expression_data.Gene.str.contains("//")].reset_index(drop=True) - multiple_gene_names = expression_data[expression_data.Gene.str.contains("//")].reset_index(drop=True) - breaks_gene_names = pd.DataFrame(columns=["Gene", "Data"]) - - for index, row in multiple_gene_names.iterrows(): - for genename in row["Gene"].split("///"): - breaks_gene_names = pd.concat( - [ - breaks_gene_names, - pd.DataFrame([{"Gene": genename, "Data": row["Data"]}]), - ], - axis=0, - ignore_index=True, - ) - - gene_expressions = pd.concat( - [single_gene_names, breaks_gene_names], axis=0, ignore_index=True - ) - gene_expressions.set_index("Gene", inplace=True) - - return gene_expressions - - -@contextlib.contextmanager -def suppress_stdout() -> Iterator[None]: - with io.StringIO() as buffer: - try: - sys.stdout = buffer - yield - finally: - sys.stdout = sys.__stdout__ diff --git a/main/como/create_context_specific_model.py b/main/como/create_context_specific_model.py index 4c45c076..c8fa22d6 100644 --- a/main/como/create_context_specific_model.py +++ b/main/como/create_context_specific_model.py @@ -1,84 +1,165 @@ +from __future__ import annotations + import argparse +import ast import collections import re +from collections.abc import Sequence +from dataclasses import dataclass from enum import Enum from pathlib import Path -from typing import Optional -from warnings import warn +from typing import NamedTuple import cobra import numpy as np +import numpy.typing as npt import pandas as pd -from cobamp.wrappers import COBRAModelObjectReader from cobra import Model from cobra.flux_analysis import pfba +from loguru import logger from troppo.methods.reconstruction.fastcore import FASTcore, FastcoreProperties from troppo.methods.reconstruction.gimme import GIMME, GIMMEProperties from troppo.methods.reconstruction.imat import IMAT, IMATProperties from troppo.methods.reconstruction.tINIT import tINIT, tINITProperties -from como.como_utilities import Compartments, split_gene_expression_data, stringlist_to_list from como.project import Config +from como.utils import Algorithm, Compartments, split_gene_expression_data, stringlist_to_list -def correct_bracket(rule: str, name: str) -> str: - """ - Correct GPR rules to format readable by - """ - rmatch = re.search(r"or|and", rule) - nmatch = re.search(r"or|and", name) - if rmatch is None or nmatch is None: - lrule = rule - lname = name.strip() - rrule = "" - rname = "" - operator = "" - else: - lrule = rule[0 : rmatch.span()[0]] - lname = name[0 : nmatch.span()[0]].strip() - rrule = rule[rmatch.span()[1] :] - rname = name[nmatch.span()[1] :] - operator = rmatch.group() - - rlist_new = [] - for ch in list(lrule): - if ch.isspace() or ch.isdigit(): # re.match(r'\w+', ch) - rlist_new.append(ch) - elif len(lname) > 0: - if ch == lname[0]: - rlist_new.append(ch) - lname = lname[1:] - rule_left = "".join(rlist_new) - - if rmatch is None: - rule_right = "" - else: - rule_right = correct_bracket(rrule, rname) - - return " ".join([rule_left, operator, rule_right]) +class _Solver(Enum): + """Solver used to seed context specific model.""" + GLPK = "GLPK" + GUROBI = "GUROBI" + SCIPY = "SCIPY" + GLPK_EXACT = "GLPK_EXACT" + + @staticmethod + def from_string(value: str) -> _Solver: + """Convert string to Solver enum.""" + match value.lower(): + case "glpk": + return _Solver.GLPK + case "gurobi": + return _Solver.GUROBI + case "scipy": + return _Solver.SCIPY + case "glpk_exact": + return _Solver.GLPK_EXACT + case _: + raise ValueError(f"Unknown solver: {value}") + + +class _BoundaryReactions(NamedTuple): + """Boundary reactions to be used in the context specific model.""" + + reactions: list[str] + lower_bounds: list[float] + upper_bounds: list[float] + + +class _BuildResults(NamedTuple): + """Results of building a context specific model.""" + + model: cobra.Model + expression_index_list: list[int] + infeasible_reactions: pd.DataFrame + + +@dataclass +class _Arguments: + """Arguments for creating a context specific model.""" + + context_name: str + reference_model: Path + active_genes_filepath: Path + objective: str + boundary_reactions_filepath: Path + exclude_reactions_filepath: Path + force_reactions_filepath: Path + recon_algorithm: Algorithm + solver: _Solver + low_threshold: int + high_threshold: int + output_filetypes: list[str] + + def __post_init__(self): + self.reference_model = Path(self.reference_model) + self.active_genes_filepath = Path(self.active_genes_filepath) + self.boundary_reactions_filepath = ( + Path(self.boundary_reactions_filepath) if self.boundary_reactions_filepath is not None else None + ) + self.exclude_reactions_filepath = ( + Path(self.exclude_reactions_filepath) if self.exclude_reactions_filepath is not None else None + ) + self.force_reactions_filepath = ( + Path(self.force_reactions_filepath) if self.force_reactions_filepath is not None else None + ) -def gene_rule_logical(expression_in: str, level: int = 0) -> str: - """ - creates an expression from GPR rule which can be evaluated as true or false - """ + if not self.reference_model.exists(): + raise FileNotFoundError(f"Reference model not found at {self.reference_model}") + if not self.active_genes_filepath.exists(): + raise FileNotFoundError(f"Active genes file not found at {self.active_genes_filepath}") + if self.boundary_reactions_filepath and not self.boundary_reactions_filepath.exists(): + raise FileNotFoundError(f"Boundary reactions file not found at {self.boundary_reactions_filepath}") + if self.exclude_reactions_filepath and not self.exclude_reactions_filepath.exists(): + raise FileNotFoundError(f"Exclude reactions file not found at {self.exclude_reactions_filepath}") + if self.force_reactions_filepath and not self.force_reactions_filepath.exists(): + raise FileNotFoundError(f"Force reactions file not found at {self.force_reactions_filepath}") + + if self.high_threshold < self.low_threshold: + raise ValueError( + f"Low threshold must be less than high threshold. " + f"Received low threshold: {self.low_threshold}, high threshold: {self.high_threshold}" + ) + + +def _correct_bracket(rule: str, name: str) -> str: + """Correct GPR rules to format readable by.""" + rule_match = re.search(r"or|and", rule) + name_match = re.search(r"or|and", name) + if rule_match is None or name_match is None: + left_rule = rule + left_name = name.strip() + right_rule = "" + right_name = "" + operator = "" + else: + left_rule = rule[: rule_match.span()[0]] + left_name = name[: name_match.span()[0]].strip() + right_rule = rule[rule_match.span()[1] :] + right_name = name[name_match.span()[1] :] + operator = rule_match.group() + + new_right_rule = [] + for char in list(left_rule): + if char.isspace() or char.isdigit(): + new_right_rule.append(char) + elif len(left_name) > 0 and char == left_name[0]: + new_right_rule.append(char) + left_name = left_name[1:] + new_left_rule = "".join(new_right_rule) + final_right_rule = "" if rule_match is None else _correct_bracket(right_rule, right_name) + return " ".join([new_left_rule, operator, final_right_rule]) + + +def _gene_rule_logical(gpr_expression: str, level: int = 0) -> str: + """Create an expression from GPR rule which can be evaluated as true or false.""" try: - loc_r = expression_in.index(")") - except BaseException: - if "and" in expression_in: - expression_in = expression_in.replace("and", ",") - expression_in = "min{" + expression_in + "}" - elif "or" in expression_in: - expression_in = expression_in.replace("or", ",") - expression_in = "max{" + expression_in + "}" + loc_r = gpr_expression.index(")") + except ValueError: + if "and" in gpr_expression: + gpr_expression = gpr_expression.replace("and", ",") + return "min{" + gpr_expression + "}" + elif "or" in gpr_expression: + gpr_expression = gpr_expression.replace("or", ",") + return "max{" + gpr_expression + "}" else: - expression_in = expression_in.replace("[", "") - expression_in = expression_in.replace("]", "") + gpr_expression = gpr_expression.replace("[", "") + return gpr_expression.replace("]", "") - return expression_in - - loc_l = expression_in[0:loc_r].rindex("(") - inner_string = expression_in[loc_l : loc_r + 1] + loc_l = gpr_expression[:loc_r].rindex("(") + inner_string = gpr_expression[loc_l : loc_r + 1] inner_string = inner_string.replace("(", "[") inner_string = inner_string.replace(")", "]") if "and" in inner_string: @@ -91,25 +172,23 @@ def gene_rule_logical(expression_in: str, level: int = 0) -> str: inner_string = inner_string.replace("[", "") inner_string = inner_string.replace("]", "") - expression_out = f"{expression_in[0:loc_l]}{inner_string}{expression_in[loc_r + 1 :]}" - expression_out = gene_rule_logical(expression_out, level + 1) + expression_out = f"{gpr_expression[:loc_l]}{inner_string}{gpr_expression[loc_r + 1:]}" + expression_out = _gene_rule_logical(expression_out, level + 1) return expression_out -def gene_rule_evaluable(expression_in: str) -> str: - """ - Make expression rule evaluable - """ - gene_reaction_by_rule = gene_rule_logical(expression_in) +def _gene_rule_evaluable(expression_in: str) -> str: + """Make expression rule evaluable.""" + gene_reaction_by_rule = _gene_rule_logical(expression_in) gene_reaction_by_rule = gene_reaction_by_rule.replace("{", "(") gene_reaction_by_rule = gene_reaction_by_rule.replace("}", ")") return gene_reaction_by_rule -def set_boundaries(model_cobra: cobra.Model, bound_rxns: list, bound_lb, bound_ub) -> tuple[cobra.Model, list]: - all_rxns = model_cobra.reactions # get all reactions +def _set_boundaries(model: cobra.Model, bound_rxns: list, bound_lb, bound_ub) -> tuple[cobra.Model, list]: + all_rxns = model.reactions # get all reactions bound_rm_rxns = [] # get boundary reactions @@ -117,76 +196,64 @@ def set_boundaries(model_cobra: cobra.Model, bound_rxns: list, bound_lb, bound_u sink_rxns = [rxn.id for rxn in all_rxns if re.search("sink_", rxn.id)] demand_rxns = [rxn.id for rxn in all_rxns if re.search("DM_", rxn.id)] - # set flag that allows all boundary reactions to be used if none are given - if len(bound_rxns) == 0: - allow_all_boundary_rxns = True - else: - allow_all_boundary_rxns = False + # Allows all boundary reactions to be used if none are given + allow_all_boundary_rxns = not bound_rxns # close sinks and demands not in boundary reactions unless no boundary reactions were given if not allow_all_boundary_rxns: for i, rxn in enumerate(sink_rxns): # set sinks to 0 if rxn not in bound_rxns: # only allow sink accumulation - getattr(model_cobra.reactions, rxn).lower_bound = 0 - getattr(model_cobra.reactions, rxn).upper_bound = 1000 + getattr(model.reactions, rxn).lower_bounds = 0 + getattr(model.reactions, rxn).upper_bounds = 1000 else: # set from file - getattr(model_cobra.reactions, rxn).lower_bound = bound_lb[i] - getattr(model_cobra.reactions, rxn).upper_bound = bound_ub[i] + getattr(model.reactions, rxn).lower_bounds = bound_lb[i] + getattr(model.reactions, rxn).upper_bounds = bound_ub[i] for i, rxn in enumerate(demand_rxns): - if rxn not in bound_rxns: # demand is one way - outside the system - getattr(model_cobra.reactions, rxn).lower_bound = 0 - getattr(model_cobra.reactions, rxn).upper_bound = 1000 - else: - getattr(model_cobra.reactions, rxn).lower_bound = 0 - getattr(model_cobra.reactions, rxn).upper_bound = bound_ub[i] + getattr(model.reactions, rxn).lower_bounds = 0 + getattr(model.reactions, rxn).upper_bounds = bound_ub[i] if rxn in bound_rxns else 0 # Reaction media - medium = model_cobra.medium # get reaction media to modify + medium = model.medium # get reaction media to modify for rxn in exchange_rxns: # open exchanges from exchange file, close unspecified exchanges if rxn not in bound_rxns: medium[rxn] = 0.0 else: medium[rxn] = -float(bound_lb[bound_rxns.index(rxn)]) - model_cobra.medium = medium # set new media + model.medium = medium # set new media - return model_cobra, bound_rm_rxns + return model, bound_rm_rxns -def feasibility_test(model_cobra: cobra.Model, step: str): +def _feasibility_test(model_cobra: cobra.Model, step: str): # check number of unsolvable reactions for reference model under media assumptions - model_cobra_rm = cobra.flux_analysis.fastcc( - model_cobra, flux_threshold=15, zero_cutoff=1e-7 - ) # create flux consistant model (rmemoves some reactions) + # create flux consistant model (rmemoves some reactions) + model_cobra_rm = cobra.flux_analysis.fastcc(model_cobra, flux_threshold=15, zero_cutoff=1e-7) incon_rxns = set(model_cobra.reactions.list_attr("id")) - set(model_cobra_rm.reactions.list_attr("id")) incon_rxns_cnt = len(incon_rxns) if step == "before_seeding": - print(f"Under given boundary assumptions, there are {str(incon_rxns_cnt)} infeasible reactions in the " "reference model.\n") - print( - "These reactions will not be considered active in context specific model construction. If any infeasible " - "reactions are found to be active according to expression data, or, are found in the force reactions " - "list, they can be found found in 'InfeasibleRxns.csv'\n" - ) - print( - "It is normal for this value to be quite large, however, if many of these reactions are active " - "according to your expression data, it is likely that you are missing some critical exchange (media) " - "reactions.\n" + logger.warning( + f"Under given boundary assumptions, there are {incon_rxns_cnt} infeasible reactions in" + f" the reference model. These reactions will not be considered active in " + f"context specific model construction. If any infeasible reactions are found to be " + f"active according to expression data, or are found in the force reactions list, " + f"they can be found found in 'InfeasibleRxns.csv'. It is normal for this value to be quite large; " + f"however, if many of these reactions are active according to your expression data, " + f"it is likely that you are missing some critical exchange (media) reactions." ) elif step == "after_seeding": - print( - f"Under given boundary assumptions, with infeasible reactions from the general model not considered " - f"there are {incon_rxns_cnt} new infeasible reactions in the context-specific model.\n" + logger.warning( + f"Under given boundary assumptions, with infeasible reactions from the general model not " + f"considered there are {incon_rxns_cnt} new infeasible reactions in the context-specific model. " + f"These reactions will be removed from the output model to ensure the model is solvable. " + f"Note that this value should be very low compared to the reference model." ) - print("These reactions will be removed from the output model to ensure the model is solvable") - print("Note that this value should be very low compared to the reference model.") - else: - pass return incon_rxns, model_cobra_rm -def seed_gimme(cobra_model, s_matrix, lb, ub, idx_objective, expr_vector): +def _build_with_gimme(cobra_model, s_matrix, lb, ub, idx_objective, expr_vector): # `Becker and Palsson (2008). Context-specific metabolic networks are # consistent with experiments. PLoS Comput. Biol. 4, e1000082.` properties = GIMMEProperties( @@ -204,7 +271,7 @@ def seed_gimme(cobra_model, s_matrix, lb, ub, idx_objective, expr_vector): context_cobra_model.remove_reactions(to_remove_ids, True) r_ids = [r.id for r in context_cobra_model.reactions] - psol = pfba(context_cobra_model) + psol = pfba(context_cobra_model) # noqa: F841 # psol = context_cobra_model.optimize() # to_remove_ids = [r_ids[r] for r in np.where(abs(psol.fluxes) < 1e-8)[0]] # context_cobra_model.remove_reactions(to_remove_ids, True) @@ -212,13 +279,16 @@ def seed_gimme(cobra_model, s_matrix, lb, ub, idx_objective, expr_vector): return context_cobra_model -def seed_fastcore(cobra_model, s_matrix, lb, ub, exp_idx_list, solver): +def _build_with_fastcore(cobra_model, s_matrix, lb, ub, exp_idx_list, solver): # 'Vlassis, Pacheco, Sauter (2014). Fast reconstruction of compact # context-specific metabolic network models. PLoS Comput. Biol. 10, # e1003424.' - warn("Fastcore requires a flux consistant model is used as refererence, " "to achieve this fastcc is required which is NOT reproducible.") - print("Creating feasible model...") - incon_rxns, cobra_model = feasibility_test(cobra_model, "other") + logger.warning( + "Fastcore requires a flux consistant model is used as refererence, " + "to achieve this fastcc is required which is NOT reproducible." + ) + logger.debug("Creating feasible model") + incon_rxns, cobra_model = _feasibility_test(cobra_model, "other") properties = FastcoreProperties(core=exp_idx_list, solver=solver) algorithm = FASTcore(s_matrix, lb, ub, properties) context_rxns = algorithm.fastcore() @@ -230,28 +300,28 @@ def seed_fastcore(cobra_model, s_matrix, lb, ub, exp_idx_list, solver): return context_cobra_model -def seed_imat(cobra_model, s_matrix, lb, ub, expr_vector, expr_thesh, idx_force, context_name): - config = Config() +def _build_with_imat( + cobra_model: cobra.Model, + s_matrix: npt.NDArray, + lb: Sequence[float], + ub: Sequence[float], + expr_vector: npt.NDArray, + expr_thesh: tuple[float, float], + force_gene_ids: Sequence[int], +) -> (cobra.Model, pd.DataFrame): expr_vector = np.array(expr_vector) - properties = IMATProperties(exp_vector=expr_vector, exp_thresholds=expr_thesh, core=idx_force, epsilon=0.01) - print("Setting properties") - algorithm = IMAT(s_matrix, lb, ub, properties) - - print("Setting algorithm") - context_rxns = algorithm.run() - - print("Running") - fluxes = algorithm.sol.to_series() - - print("Obtained flux values") + properties = IMATProperties(exp_vector=expr_vector, exp_thresholds=expr_thesh, core=force_gene_ids, epsilon=0.01) + algorithm = IMAT(s_matrix, np.array(lb), np.array(ub), properties) + context_rxns: npt.NDArray = algorithm.run() + fluxes: pd.Series = algorithm.sol.to_series() context_cobra_model = cobra_model.copy() - r_ids = [r.id for r in context_cobra_model.reactions] - pd.DataFrame({"rxns": r_ids}).to_csv(config.data_dir / "rxns_test.csv") - remove_rxns = [r_ids[int(i)] for i in range(s_matrix.shape[1]) if not np.isin(i, context_rxns)] + reaction_ids = [r.id for r in context_cobra_model.reactions] + + remove_rxns = [reaction_ids[i] for i in range(s_matrix.shape[1]) if i not in context_rxns] flux_df = pd.DataFrame(columns=["rxn", "flux"]) for idx, (_, val) in enumerate(fluxes.items()): if idx <= len(cobra_model.reactions) - 1: - r_id = str(context_cobra_model.reactions.get_by_id(r_ids[idx])).split(":")[0] + r_id = str(context_cobra_model.reactions.get_by_id(reaction_ids[idx])).split(":")[0] getattr(context_cobra_model.reactions, r_id).fluxes = val flux_df.loc[len(flux_df.index)] = [r_id, val] @@ -260,8 +330,7 @@ def seed_imat(cobra_model, s_matrix, lb, ub, expr_vector, expr_thesh, idx_force, return context_cobra_model, flux_df -def seed_tinit(cobra_model: cobra.Model, s_matrix, lb, ub, expr_vector, solver, idx_force) -> Model: - expr_vector = np.array(expr_vector) +def _build_with_tinit(cobra_model: cobra.Model, s_matrix, lb, ub, expr_vector, solver, idx_force) -> Model: properties = tINITProperties( reactions_scores=expr_vector, solver=solver, @@ -273,121 +342,135 @@ def seed_tinit(cobra_model: cobra.Model, s_matrix, lb, ub, expr_vector, solver, algorithm = tINIT(s_matrix, lb, ub, properties) algorithm.preprocessing() algorithm.build_problem() - return Model() + raise NotImplementedError("tINIT is not yet implemented") -def map_expression_to_rxn(model_cobra, gene_expression_file, recon_algorithm, low_thresh=None, high_thresh=None): - """ - Map gene ids to a reaction based on GPR (gene to protein to reaction) association rules - which are defined in general genome-scale metabolic model +def _map_expression_to_reaction( + model_cobra, + gene_expression_file, + recon_algorithm: Algorithm, + low_thresh=None, + high_thresh=None, +): + """Map gene ids to a reaction based on GPR (gene to protein to reaction) association rules. + + These rules should be defined in the general genome-scale metabolic model """ expression_data = pd.read_csv(gene_expression_file) gene_expressions = split_gene_expression_data(expression_data, recon_algorithm=recon_algorithm) expression_rxns = collections.OrderedDict() - cnt = 0 - if recon_algorithm in ["IMAT", "TINIT"]: - # unknown_val = min(gene_expressions["Data"].tolist()) + unknown_val = 1 + if recon_algorithm in {Algorithm.IMAT, Algorithm.TINIT}: unknown_val = np.mean([low_thresh, high_thresh]) # put unknowns in mid bin - elif recon_algorithm == "GIMME": + elif recon_algorithm == Algorithm.GIMME: unknown_val = -1 - elif recon_algorithm == "FASTCORE": + elif recon_algorithm == Algorithm.FASTCORE: unknown_val = 0 - else: - unknown_val = 1 - test_counter = 0 + error_count = 0 for rxn in model_cobra.reactions: - test_counter += 1 - gene_reaction_rule = correct_bracket(rxn.gene_reaction_rule, rxn.gene_name_reaction_rule) + gene_reaction_rule = _correct_bracket(rxn.gene_reaction_rule, rxn.gene_name_reaction_rule) gene_ids = re.findall(r"\d+", gene_reaction_rule) expression_rxns[rxn.id] = unknown_val if gene_reaction_rule.strip() == "": continue for gid in gene_ids: if gid in gene_expressions.index: - rep_val = " {} ".format(gene_expressions.at[gid, "Data"]) + rep_val = f' {gene_expressions.at[gid, "active"]} ' else: - rep_val = f" {str(unknown_val)} " - gene_reaction_rule = " " + gene_reaction_rule + " " # pad white space to prevent gene matches inside floats - gene_reaction_rule = gene_reaction_rule.replace(" {} ".format(gid), rep_val, 1) + rep_val = f" {unknown_val!s} " + gene_reaction_rule = f" {gene_reaction_rule} " # pad white space to prevent gene matches inside floats + gene_reaction_rule = gene_reaction_rule.replace(f" {gid} ", rep_val, 1) try: - gene_reaction_by_rule = gene_rule_evaluable(gene_reaction_rule) + gene_reaction_by_rule = _gene_rule_evaluable(gene_reaction_rule) gene_reaction_by_rule = gene_reaction_by_rule.strip() - expression_rxns[rxn.id] = eval(gene_reaction_by_rule) + expression_rxns[rxn.id] = ast.literal_eval(gene_reaction_by_rule) except BaseException: - cnt += 1 + error_count += 1 - print("Map gene expression to reactions, {} errors.".format(cnt)) + logger.info(f"Mapped gene expression to reactions, found {error_count} error(s).") expr_vector = np.array(list(expression_rxns.values()), dtype=float) return expression_rxns, expr_vector -def _create_context_specific_model( - general_model_file, - gene_expression_file, - recon_algorithm, - objective, - exclude_rxns, - force_rxns, - bound_rxns, - bound_lb, - bound_ub, - solver, - context_name, - low_thresh, - high_thresh, -): - """ - Seed a context specific model. Core reactions are determined from GPR associations with gene expression logicals. +def _build_model( # noqa: C901 + context_name: str, + general_model_file: Path, + gene_expression_file: Path, + recon_algorithm: Algorithm, + objective: str, + bound_rxns: list[str], + exclude_rxns: list[str], + force_rxns: list[str], + bound_lb: list[float], + bound_ub: list[float], + solver: str, + low_thresh: float, + high_thresh: float, +) -> _BuildResults: + """Seed a context specific reference_model. + + Core reactions are determined from GPR associations with gene expression logicals. Core reactions that do not necessarily meet GPR association requirements can be forced if in the force reaction file. Metabolite exchange (media), sinks, and demands are determined from exchanges file. Reactions can also be force excluded even if they meet GPR association requirements using the force exclude file. """ config = Config() + reference_model: cobra.Model match general_model_file.suffix: case ".mat": - cobra_model = cobra.io.load_matlab_model(general_model_file) + reference_model = cobra.io.load_matlab_model(general_model_file) case ".xml": - cobra_model = cobra.io.read_sbml_model(general_model_file) + reference_model = cobra.io.read_sbml_model(general_model_file) case ".json": - cobra_model = cobra.io.load_json_model(general_model_file) + reference_model = cobra.io.load_json_model(general_model_file) case _: - raise NameError("reference model format must be .xml, .mat, or .json") + raise NameError( + f"Reference reference_model format must be .xml, .mat, or .json, found '{general_model_file.suffix}'" + ) - cobra_model.objective = {getattr(cobra_model.reactions, objective): 1} # set objective + reference_model.objective = {getattr(reference_model.reactions, objective): 1} # set objective if objective not in force_rxns: force_rxns.append(objective) # set boundaries - cobra_model, bound_rm_rxns = set_boundaries(cobra_model, bound_rxns, bound_lb, bound_ub) + reference_model, bound_rm_rxns = _set_boundaries(reference_model, bound_rxns, bound_lb, bound_ub) # set solver - cobra_model.solver = solver.lower() + reference_model.solver = solver.lower() # check number of unsolvable reactions for reference model under media assumptions - # incon_rxns, cobra_model = feasibility_test(cobra_model, "before_seeding") + # incon_rxns, cobra_model = _feasibility_test(cobra_model, "before_seeding") incon_rxns = [] - # CoBAMP methods - cobamp_model = COBRAModelObjectReader(cobra_model) # load model in readable format for CoBAMP - cobamp_model.get_irreversibilities(True) - s_matrix = cobamp_model.get_stoichiometric_matrix() - lb, ub = cobamp_model.get_model_bounds(False, True) - rx_names = cobamp_model.get_reaction_and_metabolite_ids()[0] + s_matrix = cobra.util.array.create_stoichiometric_matrix(reference_model, array_type="dense") + lb = [] + ub = [] + rx_names = [] + for reaction in reference_model.reactions: + lb.append(reaction.lower_bound) + ub.append(reaction.upper_bound) + rx_names.append(reaction.id) # get expressed reactions - expression_rxns, expr_vector = map_expression_to_rxn( - cobra_model, gene_expression_file, recon_algorithm, high_thresh=high_thresh, low_thresh=low_thresh + expression_rxns, expr_vector = _map_expression_to_reaction( + reference_model, + gene_expression_file, + recon_algorithm, + high_thresh=high_thresh, + low_thresh=low_thresh, ) - # find reactions in the force reactions file that are not in general model and warn user for rxn in force_rxns: if rxn not in rx_names: - warn(f"{rxn} from force reactions not in general model, check BiGG (or relevant database used in your " "general model) for synonyms") + logger.warning( + f"The force reaction '{rxn}' was not found in the general reference_model. " + f"Check BiGG, or the relevant database for your general reference_model, for synonyms." + ) # collect list of reactions that are infeasible but active in expression data or user defined infeas_exp_rxns = [] @@ -395,7 +478,7 @@ def _create_context_specific_model( infeas_exp_cnt = 0 infeas_force_cnt = 0 - for idx, (rxn, exp) in enumerate(expression_rxns.items()): + for idx, rxn in enumerate(expression_rxns): # log reactions in expressed and force lists that are infeasible that the user may wish to review if rxn in incon_rxns and expr_vector[idx] == 1: infeas_exp_cnt += 1 @@ -405,259 +488,216 @@ def _create_context_specific_model( infeas_force_rxns.append(rxn) # make changes to expressed reactions base on user defined force/exclude reactions - # TODO: if not using bound reactions file, add two sets of exchange reactoins to be put in either low or mid bin + # TODO: if not using bound reactions file, add two sets of exchange reactions to be put in either low or mid bin if rxn in force_rxns: - expr_vector[idx] = high_thresh + 0.1 if recon_algorithm in ["TINIT", "IMAT"] else 1 + expr_vector[idx] = high_thresh + 0.1 if recon_algorithm.value in {"TINIT", "IMAT"} else 1 if rxn in incon_rxns or rxn in exclude_rxns: - expr_vector[idx] = low_thresh - 0.1 if recon_algorithm in ["TINIT", "IMAT"] else 0 + expr_vector[idx] = low_thresh - 0.1 if recon_algorithm.value in {"TINIT", "IMAT"} else 0 idx_obj = rx_names.index(objective) idx_force = [rx_names.index(rxn) for rxn in force_rxns if rxn in rx_names] - exp_idx_list = [idx for (idx, val) in enumerate(expr_vector) if val > 0] + exp_idx_list = [i for (i, val) in enumerate(expr_vector) if val > 0] # type: ignore exp_thresh = (low_thresh, high_thresh) - # switch case dictionary runs the functions making it too slow, better solution then elif ladder? - if recon_algorithm == "GIMME": - context_model_cobra = seed_gimme(cobra_model, s_matrix, lb, ub, idx_obj, expr_vector) - elif recon_algorithm == "FASTCORE": - context_model_cobra = seed_fastcore(cobra_model, s_matrix, lb, ub, exp_idx_list, solver) - elif recon_algorithm == "IMAT": - flux_df: pd.DataFrame + if recon_algorithm == Algorithm.GIMME: + context_model_cobra = _build_with_gimme(reference_model, s_matrix, lb, ub, idx_obj, expr_vector) + elif recon_algorithm == Algorithm.FASTCORE: + context_model_cobra = _build_with_fastcore(reference_model, s_matrix, lb, ub, exp_idx_list, solver) + elif recon_algorithm == Algorithm.IMAT: context_model_cobra: cobra.Model - context_model_cobra, flux_df = seed_imat( - cobra_model, - s_matrix, - lb, - ub, - expr_vector, - exp_thresh, - idx_force, - context_name, + context_model_cobra, flux_df = _build_with_imat( + reference_model, s_matrix, lb, ub, expr_vector, exp_thresh, idx_force ) imat_reactions = flux_df.rxn model_reactions = [reaction.id for reaction in context_model_cobra.reactions] reaction_intersections = set(imat_reactions).intersection(model_reactions) flux_df = flux_df[~flux_df["rxn"].isin(reaction_intersections)] - flux_df.to_csv(config.data_dir / "results" / context_name / f"{recon_algorithm}_flux.csv") - - elif recon_algorithm == "TINIT": - context_model_cobra = seed_tinit(cobra_model, s_matrix, lb, ub, expr_vector, solver, idx_force) - else: - raise ValueError(f"Unsupported reconstruction algorithm: {recon_algorithm}. Must be 'IMAT', 'GIMME', or 'FASTCORE'") + flux_df.to_csv(config.data_dir / "results" / context_name / f"{recon_algorithm.value}_flux.csv") + elif recon_algorithm == Algorithm.TINIT: + context_model_cobra = _build_with_tinit(reference_model, s_matrix, lb, ub, expr_vector, solver, idx_force) - # check number of unsolvable reactions for seeded model under media assumptions - # incon_rxns_cs, context_model_cobra = feasibility_test(context_model_cobra, "after_seeding") incon_rxns_cs = [] - - # if recon_algorithm in ["IMAT"]: - # final_rxns = [rxn.id for rxn in context_model_cobra.reactions] - # imat_rxns = flux_df.rxn - # for rxn in imat_rxns: - # if rxn not in final_rxns: - # flux_df = flux_df[flux_df.rxn != rxn] - # - incon_df = pd.DataFrame({"general_infeasible_rxns": list(incon_rxns)}) infeas_exp_df = pd.DataFrame({"expressed_infeasible_rxns": infeas_exp_rxns}) infeas_force_df = pd.DataFrame({"infeasible_rxns_in_force_list": infeas_exp_rxns}) incon_df_cs = pd.DataFrame({"infeasible_rxns_from_seeding": list(incon_rxns_cs)}) - infeasible_df = pd.concat( - [incon_df, infeas_exp_df, infeas_force_df, incon_df_cs], - ignore_index=True, - axis=1, - ) - infeasible_df.columns = [ - "InfeasRxns", - "ExpressedInfeasRxns", - "ForceInfeasRxns", - "ContextInfeasRxns", - ] + infeasible_df = pd.concat([incon_df, infeas_exp_df, infeas_force_df, incon_df_cs], ignore_index=True, axis=1) + infeasible_df.columns = ["InfeasRxns", "ExpressedInfeasRxns", "ForceInfeasRxns", "ContextInfeasRxns"] - return context_model_cobra, exp_idx_list, infeasible_df + return _BuildResults( + model=context_model_cobra, + expression_index_list=exp_idx_list, + infeasible_reactions=infeasible_df, + ) -class Algorithm(Enum): - GIMME = "GIMME" - FASTCORE = "FASTCORE" - IMAT = "IMAT" - TINIT = "TINIT" +def _create_df(path: Path) -> pd.DataFrame: + match path.suffix: + case ".csv": + df = pd.read_csv(path, header=0, sep=",") + case ".tsv": + df = pd.read_csv(path, header=0, sep="\t") + case ".xlsx" | ".xls": + df = pd.read_excel(path, header=0) + case _: + raise FileNotFoundError(f"File not found! Must be a csv, tsv, or Excel file. Searching for: {path}") + df.columns = [c.lower() for c in df.columns] + return df + + +def _collect_boundary_reactions(path: Path) -> _BoundaryReactions: + df = _create_df(path) + for column in df.columns: + if column not in [ + "reaction", + "abbreviation", + "compartment", + "minimum reaction rate", + "maximum reaction rate", + ]: + raise ValueError( + f"Boundary reactions file must have columns named 'Reaction', 'Abbreviation', 'Compartment', " + f"'Minimum Reaction Rate', and 'Maximum Reaction Rate'. Found: {column}" + ) + + reactions: list[str] = [] + boundary_type: list[str] = df["reaction"].tolist() + reaction_abbreviation: list[str] = df["abbreviation"].tolist() + reaction_compartment: list[str] = df["compartment"].tolist() + lower_bounds = df["minimum reaction rate"].tolist() + upper_bounds = df["maximum reaction rate"].tolist() + for i in range(len(boundary_type)): + current_type: str = boundary_type[i] + temp_reaction: str = "" + + match current_type.lower(): + case "exchange": + temp_reaction += "EX_" + case "demand": + temp_reaction += "DM_" + case "sink": + temp_reaction += "SK_" + + shorthand_compartment = Compartments.get(reaction_compartment[i]) + temp_reaction += f"{reaction_abbreviation[i]}[{shorthand_compartment}]" + reactions.append(temp_reaction) + return _BoundaryReactions( + reactions=reactions, + lower_bounds=lower_bounds, + upper_bounds=upper_bounds, + ) -class Solver(Enum): - GLPK = "GLPK" - GUROBI = "GUROBI" +def _write_model_to_disk( + model: cobra.Model, + output_directory: Path, + context_name: str, + output_filetypes: list[str], + algorithm: Algorithm, +) -> None: + if "mat" in output_filetypes: + cobra.io.save_matlab_model(model, output_directory / f"{context_name}_SpecificModel_{algorithm.value}.mat") + if "xml" in output_filetypes: + cobra.io.write_sbml_model(model, output_directory / f"{context_name}_SpecificModel_{algorithm.value}.xml") + if "json" in output_filetypes: + cobra.io.save_json_model(model, output_directory / f"{context_name}_SpecificModel_{algorithm.value}.json") -def create_context_specific_model( +def create_context_specific_model( # noqa: C901 context_name: str, reference_model: Path, genes_file: Path, objective: str = "biomass_reaction", - boundary_rxns_filepath: Optional[str] = None, - exclude_rxns_filepath: Optional[str] = None, - force_rxns_filepath: Optional[str] = None, + boundary_rxns_filepath: str | Path | None = None, + exclude_rxns_filepath: str | Path | None = None, + force_rxns_filepath: str | Path | None = None, algorithm: Algorithm = Algorithm.GIMME, low_threshold: float = -5, high_threshold: float = -3, - solver: Solver = Solver.GLPK, - output_filetypes: list[str] = ["mat"], + solver: _Solver = _Solver.GLPK, + output_filetypes: list[str] | None = None, ): + """Create a context-specific model using the provided data.""" if not reference_model.exists(): raise FileNotFoundError(f"Reference model not found at {reference_model}") if not genes_file.exists(): raise FileNotFoundError(f"Active genes file not found at {genes_file}") + if output_filetypes is None: + output_filetypes = ["mat"] - config = Config() - print(f"Active Genes: {genes_file}") + for output_type in output_filetypes: + if output_type not in {"xml", "mat", "json"}: + raise ValueError(f"Output file type {output_type} not recognized. Must be one of: 'xml', 'mat', 'json'") - boundary_rxns = [] - boundary_rxns_upper: list[float] = [] - boundary_rxns_lower: list[float] = [] + if algorithm not in Algorithm: + raise ValueError(f"Algorithm {algorithm} not supported. Please use one of: GIMME, FASTCORE, or IMAT") + + if solver not in _Solver: + raise ValueError(f"Solver '{solver}' not supported. Use 'GLPK' or 'GUROBI'") if boundary_rxns_filepath: - boundary_rxns_filepath: Path = Path(boundary_rxns_filepath) + boundary_reactions = _collect_boundary_reactions(boundary_rxns_filepath) - print(f"Boundary Reactions: {str(boundary_rxns_filepath)}") - if boundary_rxns_filepath.suffix == ".csv": - df: pd.DataFrame = pd.read_csv(boundary_rxns_filepath, header=0, sep=",") - elif boundary_rxns_filepath.suffix in [".xlsx", ".xls"]: - df: pd.DataFrame = pd.read_excel(boundary_rxns_filepath, header=0) - else: - raise FileNotFoundError(f"Boundary reactions file not found! Must be a csv or Excel file. Searching for: {boundary_rxns_filepath}") - - # convert all columns to lowercase - df.columns = [column.lower() for column in df.columns] - - # Make sure the columns are named correctly. They should be "Reaction", "Abbreviation", "Compartment", "Minimum Reaction Rate", and "Maximum Reaction Rate" - for column in df.columns: - if column not in ["reaction", "abbreviation", "compartment", "minimum reaction rate", "maximum reaction rate"]: - raise ValueError( - f"Boundary reactions file must have columns named 'Reaction', 'Abbreviation', 'Compartment', 'Minimum Reaction Rate', and 'Maximum Reaction Rate'. Found: {column}" - ) - - reaction_type: list[str] = df["reaction"].tolist() - reaction_abbreviation: list[str] = df["abbreviation"].tolist() - reaction_compartment: list[str] = df["compartment"].tolist() - boundary_rxns_lower = df["minimum reaction rate"].tolist() - boundary_rxns_upper = df["maximum reaction rate"].tolist() - - reaction_formula: list[str] = [] - for i in range(len(reaction_type)): - current_type: str = reaction_type[i] - temp_reaction: str = "" - - match current_type.lower(): - case "exchange": - temp_reaction += "EX_" - case "demand": - temp_reaction += "DM_" - case "sink": - temp_reaction += "SK_" - - shorthand_compartment = Compartments.get(reaction_compartment[i]) - temp_reaction += f"{reaction_abbreviation[i]}[{shorthand_compartment}]" - boundary_rxns.append(temp_reaction) - # reaction_formula.append(temp_reaction) - - del df - - exclude_rxns = [] + exclude_rxns: list[str] = [] if exclude_rxns_filepath: exclude_rxns_filepath: Path = Path(exclude_rxns_filepath) + df = _create_df(exclude_rxns_filepath) + if "abbreviation" not in df.columns: + raise ValueError("The exclude reactions file should have a single column with a header named Abbreviation") + exclude_rxns = df["abbreviation"].tolist() - print(f"Reading {exclude_rxns_filepath} for exclude reactions") - if exclude_rxns_filepath.suffix == ".csv": - df = pd.read_csv(exclude_rxns_filepath, header=0) - elif exclude_rxns_filepath.suffix == ".xlsx" or exclude_rxns_filepath.suffix == ".xls": - df = pd.read_excel(exclude_rxns_filepath, header=0) - else: - raise ValueError("exclude_rxns_filepath must be a path to a csv or xlsx file with one column.") - - exclude_rxns = df["Abbreviation"].tolist() - - force_rxns = [] + force_rxns: list[str] = [] if force_rxns_filepath: force_rxns_filepath: Path = Path(force_rxns_filepath) - - print(f"Force Reactions: {force_rxns_filepath}") - if force_rxns_filepath.suffix == ".csv": - df = pd.read_csv(force_rxns_filepath, header=0) - elif force_rxns_filepath.suffix == ".xlsx" or force_rxns_filepath.suffix == ".xls": - df = pd.read_excel(force_rxns_filepath, header=0) - else: - raise ValueError("--force-reactions-file must be a path to a csv or xlsx file with one column.") - force_rxns = df["Abbreviation"].tolist() - - # Assert output types are valid - for output_type in output_filetypes: - if output_type not in ["xml", "mat", "json"]: - raise ValueError(f"Output file type {output_type} not recognized. Must be one of: 'xml', 'mat', 'json'") - - if algorithm not in Algorithm: - raise ValueError(f"Algorithm {algorithm} not supported. Please use one of: GIMME, FASTCORE, or IMAT") - - if solver not in Solver: - print(f"Solver {solver} not supported. Please use 'GLPK' or 'GUROBI'") - - print(f"Creating '{context_name}' model using '{algorithm}' reconstruction and '{solver}' solver") - context_model, core_list, infeas_df = _create_context_specific_model( - reference_model, - genes_file, - algorithm.value, - objective, - exclude_rxns, - force_rxns, - boundary_rxns, - boundary_rxns_lower, - boundary_rxns_upper, - solver.value, - context_name, - low_threshold, - high_threshold, + df = _create_df(force_rxns_filepath) + if "abbreviation" not in df.columns: + raise ValueError("The force reactions file should have a single column with a header named Abbreviation") + force_rxns = df["abbreviation"].tolist() + + logger.info(f"Creating '{context_name}' model using '{algorithm.value}' reconstruction and '{solver.value}' solver") + build_results: _BuildResults = _build_model( + context_name=context_name, + general_model_file=reference_model, + gene_expression_file=genes_file, + recon_algorithm=algorithm, + objective=objective, + bound_rxns=boundary_reactions.reactions, + bound_lb=boundary_reactions.lower_bounds, + bound_ub=boundary_reactions.upper_bounds, + exclude_rxns=exclude_rxns, + force_rxns=force_rxns, + solver=solver.value, + low_thresh=low_threshold, + high_thresh=high_threshold, ) - infeas_df.to_csv( - config.result_dir / context_name / f"{context_name}_infeasible_rxns.csv", - index=False, + config = Config() + build_results.infeasible_reactions.to_csv( + config.result_dir / context_name / f"{context_name}_infeasible_rxns.csv", index=False ) if algorithm == Algorithm.FASTCORE: - pd.DataFrame(core_list).to_csv( - config.result_dir / context_name / f"{context_name}_core_rxns.csv", - index=False, + pd.DataFrame(build_results.expression_index_list).to_csv( + config.result_dir / context_name / f"{context_name}_core_rxns.csv", index=False ) output_directory = config.result_dir / context_name - if "mat" in output_filetypes: - cobra.io.save_matlab_model( - context_model, - output_directory / f"{context_name}_SpecificModel_{algorithm.value}.mat", - ) - if "xml" in output_filetypes: - cobra.io.write_sbml_model( - context_model, - output_directory / f"{context_name}_SpecificModel_{algorithm.value}.xml", - ) - if "json" in output_filetypes: - cobra.io.save_json_model( - context_model, - output_directory / f"{context_name}_SpecificModel_{algorithm.value}.json", - ) - - print("") - print(f"Saved output file to {output_directory}") - print(f"Number of Genes: {len(context_model.genes):,}") - print(f"Number of Metabolites: {len(context_model.metabolites):,}") - print(f"Number of Reactions: {len(context_model.reactions):,}") - print("\nModel successfully created!") - + _write_model_to_disk( + model=build_results.model, + output_directory=output_directory, + context_name=context_name, + output_filetypes=output_filetypes, + algorithm=algorithm, + ) -def print_filetype_help(): - print("Unsupported model format. Current support is for: 'xml', 'mat', and 'json'." "Or use multiple with: 'xml mat json'") + logger.success(f"Saved output file to {output_directory}") + logger.info(f"Number of Genes: {len(build_results.model.genes):,}") + logger.info(f"Number of Metabolites: {len(build_results.model.metabolites):,}") + logger.info(f"Number of Reactions: {len(build_results.model.reactions):,}") -def parse_args(): +def _parse_args(): parser = argparse.ArgumentParser( prog="create_context_specific_model.py", description="Seed a context-specific model from a list of expressed genes, a reference", @@ -677,7 +717,7 @@ def parse_args(): "--reference-model-filepath", type=str, required=True, - dest="modelfile", + dest="reference_model", help="Name of Genome-scale metabolic model to seed the context model to. For example, the " "GeneralModelUpdatedV2.mat, is a modified Recon3D model. We also provide iMM_madrid for mouse." "OT can be .mat, .xml, or .json.", @@ -687,7 +727,7 @@ def parse_args(): "--active-genes-filepath", type=str, required=True, - dest="genefile", + dest="active_genes_filepath", help="Path to logical table of active genes output from merge_xomics.py called " "ActiveGenes_contextName_Merged.csv. Should be in the corresponding context/context folder " "inside /main/data/results/contextName/. The json file output from the function using " @@ -697,7 +737,6 @@ def parse_args(): "-o", "--objective", type=str, - required=False, default="biomass_reaction", dest="objective", help="Reaction ID of the objective function in the model. Generally a biomass function.", @@ -706,9 +745,8 @@ def parse_args(): "-b", "--boundary-reactions-filepath", type=str, - required=False, default=None, - dest="boundary_rxns_filepath", + dest="boundary_reactions_filepath", help="Path to file contains the exchange (media), sink, and demand reactions which " "the model should use to fulfill the reactions governed by transcriptomic and proteomics " "data inputs. It must be a csv or xlsx with three columns: Rxn, Lowerbound, Upperbound. If not " @@ -719,9 +757,8 @@ def parse_args(): "-x", "--exclude-reactions-filepath", type=str, - required=False, default=None, - dest="exclude_rxns_filepath", + dest="exclude_reactions_filepath", help="Filepath to file that contains reactions which will be removed from active reactions " "the model to use when seeding, even if considered active from transcriptomic and " "proteomics data inputs. It must be a csv or xlsx with one column of reaction IDs consistent with " @@ -731,9 +768,8 @@ def parse_args(): "-f", "--force-reactions-filepath", type=str, - required=False, default=None, - dest="force_rxns_filepath", + dest="force_reactions_filepath", help="Filepath to file that contains reactions which will be added to active reactions for " "the model to use when seeding (unless it causes the model to be unsolvable), regardless " "of results of transcriptomic and proteomics data inputs. It must be a csv or xlsx with one " @@ -743,22 +779,31 @@ def parse_args(): "-a", "--algorithm", type=str, - required=False, default="GIMME", - dest="algorithm", - help="Algorithm used to seed context specific model to the Genome-scale model. Can be either " "GIMME, FASTCORE, iMAT, or tINIT.", + dest="recon_algorithm", + help="Algorithm used to seed context specific model to the Genome-scale model. " + "Can be either GIMME, FASTCORE, iMAT, or tINIT.", ) parser.add_argument( - "-lt", "--low-threshold", type=float, required=False, default=-5, dest="low_threshold", help="Low to mid bin cutoff for iMAT solution" + "-lt", + "--low-threshold", + type=float, + default=-5, + dest="low_threshold", + help="Low to mid bin cutoff for iMAT solution", ) parser.add_argument( - "-ht", "--high-threshold", type=float, required=False, default=-3, dest="high_threshold", help="Mid to high bin cutoff for iMAT solution" + "-ht", + "--high-threshold", + type=float, + default=-3, + dest="high_threshold", + help="Mid to high bin cutoff for iMAT solution", ) parser.add_argument( "-s", "--solver", type=str, - required=False, default="glpk", dest="solver", help="Solver used to seed model and attempt to solve objective. Default is GLPK, also takes " @@ -771,7 +816,6 @@ def parse_args(): "--output-filetypes", type=str, nargs="+", - required=False, default="mat", dest="output_filetypes", help="Filetypes to save seeded model type. Can be either a string with one filetype such as " @@ -780,179 +824,25 @@ def parse_args(): "Note the outer quotes required to be interpreted by cmd. This a string, not a python list", ) args = parser.parse_args() - return args - - -def main(): - """ - Seed a context-specific model from a list of expressed genes, a reference - """ - args = parse_args() - - context_name: str = args.context_name - reference_model: Path = Path(args.modelfile) - genefile: Path = Path(args.genefile) - objective: str = args.objective - boundary_rxns_filepath = args.boundary_rxns_filepath - exclude_rxns_filepath = args.exclude_rxns_filepath - force_rxns_filepath = args.force_rxns_filepath - recon_alg = args.algorithm.upper() - low_threshold = args.low_threshold - high_threshold = args.high_threshold - solver = args.solver.upper() - output_filetypes = stringlist_to_list(args.output_filetypes) - - if not reference_model.exists(): - raise FileNotFoundError(f"Reference model not found at {reference_model}") - - if not genefile.exists(): - raise FileNotFoundError(f"Active genes file not found at {genefile}") - - print(f"Active Genes: {genefile}") - - boundary_rxns = [] - boundary_rxns_upper: list[float] = [] - boundary_rxns_lower: list[float] = [] - - if boundary_rxns_filepath: - boundary_rxns_filepath: Path = Path(boundary_rxns_filepath) - - print(f"Boundary Reactions: {str(boundary_rxns_filepath)}") - if boundary_rxns_filepath.suffix == ".csv": - df: pd.DataFrame = pd.read_csv(boundary_rxns_filepath, header=0, sep=",") - elif boundary_rxns_filepath.suffix in [".xlsx", ".xls"]: - df: pd.DataFrame = pd.read_excel(boundary_rxns_filepath, header=0) - else: - raise FileNotFoundError(f"Boundary reactions file not found! Must be a csv or Excel file. Searching for: {boundary_rxns_filepath}") - - # convert all columns to lowercase - df.columns = [column.lower() for column in df.columns] - - # Make sure the columns are named correctly. They should be "Reaction", "Abbreviation", "Compartment", "Minimum Reaction Rate", and "Maximum Reaction Rate" - for column in df.columns: - if column not in ["reaction", "abbreviation", "compartment", "minimum reaction rate", "maximum reaction rate"]: - raise ValueError( - f"Boundary reactions file must have columns named 'Reaction', 'Abbreviation', 'Compartment', 'Minimum Reaction Rate', and 'Maximum Reaction Rate'. Found: {column}" - ) - - reaction_type: list[str] = df["reaction"].tolist() - reaction_abbreviation: list[str] = df["abbreviation"].tolist() - reaction_compartment: list[str] = df["compartment"].tolist() - boundary_rxns_lower = df["minimum reaction rate"].tolist() - boundary_rxns_upper = df["maximum reaction rate"].tolist() - - reaction_formula: list[str] = [] - for i in range(len(reaction_type)): - current_type: str = reaction_type[i] - temp_reaction: str = "" - - match current_type.lower(): - case "exchange": - temp_reaction += "EX_" - case "demand": - temp_reaction += "DM_" - case "sink": - temp_reaction += "SK_" - - shorthand_compartment = Compartments.get(reaction_compartment[i]) - temp_reaction += f"{reaction_abbreviation[i]}[{shorthand_compartment}]" - boundary_rxns.append(temp_reaction) - # reaction_formula.append(temp_reaction) - - del df - - exclude_rxns = [] - if exclude_rxns_filepath: - exclude_rxns_filepath: Path = Path(exclude_rxns_filepath) - if not exclude_rxns_filepath.exists(): - raise FileNotFoundError(f"Exclude reactions file not found at {exclude_rxns_filepath}") - if exclude_rxns_filepath.suffix not in [".csv", ".xlsx", ".xls"]: - raise FileNotFoundError(f"Exclude reactions file not found! Must be a csv or Excel file. Searching for: {exclude_rxns_filepath}") - - print(f"Reading {exclude_rxns_filepath} for exclude reactions") - df = pd.DataFrame() - if exclude_rxns_filepath.suffix == ".csv": - df = pd.read_csv(exclude_rxns_filepath, header=0) - elif exclude_rxns_filepath.suffix == ".xlsx" or exclude_rxns_filepath.suffix == ".xls": - df = pd.read_excel(exclude_rxns_filepath, header=0) - if "Abbreviation" not in df.columns: - raise ValueError("Exclude reactions file must have a column called 'Abbreviation'") - - exclude_rxns = df["Abbreviation"].tolist() - - force_rxns = [] - if force_rxns_filepath: - force_rxns_filepath: Path = Path(force_rxns_filepath) - if not force_rxns_filepath.exists(): - raise FileNotFoundError(f"Force reactions file not found at {force_rxns_filepath}") - if force_rxns_filepath.suffix not in [".csv", ".xlsx", ".xls"]: - raise FileNotFoundError(f"Force reactions file not found! Must be a csv or Excel file. Searching for: {force_rxns_filepath}") - print(f"Force Reactions: {force_rxns_filepath}") - - df = pd.DataFrame() - if force_rxns_filepath.suffix == ".csv": - df = pd.read_csv(force_rxns_filepath, header=0) - elif force_rxns_filepath.suffix == ".xlsx" or force_rxns_filepath.suffix == ".xls": - df = pd.read_excel(force_rxns_filepath, header=0) - if "Abbreviation" not in df.columns: - raise ValueError("Force reactions file must have a column called 'Abbreviation'") - force_rxns = df["Abbreviation"].tolist() - - config = Config() - - # Assert output types are valid - for output_type in output_filetypes: - if output_type not in ["xml", "mat", "json"]: - raise ValueError("Output file type must be one of: xml, mat, json") - - if recon_alg not in ["FASTCORE", "GIMME", "IMAT"]: - raise ValueError("Recon algorithm must be one of: FASTCORE, GIMME, IMAT") - - if solver not in ["GUROBI", "GLPK"]: - raise ValueError("Solver must be one of: GUROBI, GLPK") - - print(f"Creating '{context_name}' model using '{recon_alg}' reconstruction and '{solver}' solver") - context_model, core_list, infeas_df = _create_context_specific_model( - reference_model, - genefile, - recon_alg, - objective, - exclude_rxns, - force_rxns, - boundary_rxns, - boundary_rxns_lower, - boundary_rxns_upper, - solver, - context_name, - low_threshold, - high_threshold, - ) - - infeas_df.to_csv( - config.result_dir / context_name / f"{context_name}_infeasible_rxns.csv", - index=False, - ) - - if recon_alg == "FASTCORE": - pd.DataFrame(core_list).to_csv( - config.result_dir / context_name / f"{context_name}_core_rxns.csv", - index=False, - ) - - output_directory = config.result_dir / context_name - if "mat" in output_filetypes: - cobra.io.save_matlab_model(context_model, output_directory / f"{context_name}_SpecificModel_{recon_alg}.mat") - if "xml" in output_filetypes: - cobra.io.write_sbml_model(context_model, output_directory / f"{context_name}_SpecificModel_{recon_alg}.xml") - if "json" in output_filetypes: - cobra.io.save_json_model(context_model, output_directory / f"{context_name}_SpecificModel_{recon_alg}.json") - - print("\nModel successfully created!") - print(f"Saved output file to {output_directory}") - print(f"Number of Genes: {len(context_model.genes):,}") - print(f"Number of Metabolites: {len(context_model.metabolites):,}") - print(f"Number of Reactions: {len(context_model.reactions):,}") + args.output_filetypes = stringlist_to_list(args.output_filetypes) + args.solver = _Solver.from_string(args.solver) # type: ignore + args.recon_algorithm = Algorithm.from_string(args.recon_algorithm) # type: ignore + return _Arguments(**vars(args)) if __name__ == "__main__": - main() + args = _parse_args() + create_context_specific_model( + context_name=args.context_name, + reference_model=args.reference_model, + genes_file=args.active_genes_filepath, + objective=args.objective, + boundary_rxns_filepath=args.boundary_reactions_filepath, + exclude_rxns_filepath=args.exclude_reactions_filepath, + force_rxns_filepath=args.force_reactions_filepath, + algorithm=args.recon_algorithm, + low_threshold=args.low_threshold, + high_threshold=args.high_threshold, + solver=args.solver, + output_filetypes=args.output_filetypes, + ) diff --git a/main/como/custom_types.py b/main/como/custom_types.py new file mode 100644 index 00000000..a4fcd19b --- /dev/null +++ b/main/como/custom_types.py @@ -0,0 +1,23 @@ +from enum import Enum + + +class RNASeqPreparationMethod(Enum): + TOTAL = "total" + MRNA = "mrna" + SCRNA = "scrna" + + @staticmethod + def from_string(value: str) -> "RNASeqPreparationMethod": + """Build a preparation method object from a string.""" + match_value = "".join(c for c in value if c.isascii()).lower() + + match match_value: + case "total" | "trna": + return RNASeqPreparationMethod.TOTAL + case "mrna": + return RNASeqPreparationMethod.MRNA + case "scrna": + return RNASeqPreparationMethod.SCRNA + case _: + possible_values = [t.value for t in RNASeqPreparationMethod] + raise ValueError(f"Filtering technique must be one of {possible_values}; got: {value}") diff --git a/main/como/disease_analysis.py b/main/como/disease_analysis.py index db0b17c1..8eb7b52e 100644 --- a/main/como/disease_analysis.py +++ b/main/como/disease_analysis.py @@ -1,4 +1,6 @@ -#!/usr/bin/python3 +# ruff: noqa + + import argparse import json from pathlib import Path diff --git a/main/como/knock_out_simulation.py b/main/como/knock_out_simulation.py index 57367493..fe96c784 100644 --- a/main/como/knock_out_simulation.py +++ b/main/como/knock_out_simulation.py @@ -1,9 +1,10 @@ +# ruff: noqa + import argparse -from concurrent.futures import Future, as_completed, ProcessPoolExecutor, ThreadPoolExecutor import os import re import sys -from concurrent.futures import Future, ProcessPoolExecutor, as_completed +from concurrent.futures import Future, ProcessPoolExecutor, ThreadPoolExecutor, as_completed from dataclasses import dataclass from pathlib import Path from typing import Union @@ -32,9 +33,7 @@ def _perform_knockout( gene_id: str, reference_solution, ) -> tuple[str, pd.Series]: - """ - This function will perform a single gene knockout. It will be used in multiprocessing - """ + """This function will perform a single gene knockout. It will be used in multiprocessing""" with model: gene: cobra.Gene = model.genes.get_by_id(gene_id) gene.knock_out() @@ -119,12 +118,12 @@ def knock_out_simulation( flux_solution_diffs = flux_solution.sub(wild_type_model["fluxes"], axis=0) return KnockoutResults( - model, - gene_ind2genes, - genes_with_metabolic_effects, - flux_solution, - flux_solution_ratios, - flux_solution_diffs, + model=model, + gene_ind2genes=gene_ind2genes, + genes_with_metabolic_effects=genes_with_metabolic_effects, + flux_solution=flux_solution, + flux_solution_ratios=flux_solution_ratios, + flux_solution_diffs=flux_solution_diffs, ) @@ -190,7 +189,7 @@ def score_gene_pairs(gene_pairs, filename, input_reg): d_score.at[p_gene, "score"] = d_s d_score.index.name = "Gene ID" - d_score.to_csv(os.path.join(configs.data_dir, filename)) + d_score.to_csv(configs.data_dir / filename) return d_score @@ -281,7 +280,8 @@ def main(argv): parser = argparse.ArgumentParser( prog="knock_out_simulation.py", description="This script is responsible for mapping drug targets in metabolic models, performing knock out simulations, and comparing simulation results with disease genes. It also identified drug targets and repurposable drugs.", - epilog="For additional help, please post questions/issues in the MADRID GitHub repo at " "https://github.com/HelikarLab/COMO", + epilog="For additional help, please post questions/issues in the MADRID GitHub repo at " + "https://github.com/HelikarLab/COMO", ) parser.add_argument( "-m", @@ -382,7 +382,7 @@ def main(argv): output_dir = Path(configs.data_dir, "results", context, disease) inhibitors_filepath = Path(output_dir, f"{context}_{disease}_inhibitors.tsv") - biodbnet = BioDBNet(show_progress=True, cache=False) + biodbnet = BioDBNet(cache=False) thread_pool = ThreadPoolExecutor(max_workers=1) print(f"Output directory: '{output_dir.as_posix()}'") @@ -416,14 +416,7 @@ def main(argv): cobra_model.solver = solver thread_pool.shutdown() - ( - model, - gene_ind2genes, - has_effects_gene, - fluxsolution, - flux_solution_ratios, - flux_solution_diffs, - ) = knock_out_simulation( + knockout_results = knock_out_simulation( model=cobra_model, inhibitors_filepath=inhibitors_filepath, drug_db=drug_info_df, @@ -437,24 +430,24 @@ def main(argv): gene_pairs_down = create_gene_pairs( configs.data_dir, - model, - gene_ind2genes, - fluxsolution, - flux_solution_ratios, - flux_solution_diffs, - has_effects_gene, + knockout_results.model, + knockout_results.gene_ind2genes, + knockout_results.flux_solution, + knockout_results.flux_solution_ratios, + knockout_results.flux_solution_diffs, + knockout_results.genes_with_metabolic_effects, disease_genes_filename=disease_down_file, ) gene_pairs_down.to_csv(os.path.join(output_dir, f"{context}_Gene_Pairs_Inhi_Fratio_DOWN.txt"), index=False) gene_pairs_up = create_gene_pairs( configs.data_dir, - model, - gene_ind2genes, - fluxsolution, - flux_solution_ratios, - flux_solution_diffs, - has_effects_gene, + knockout_results.model, + knockout_results.gene_ind2genes, + knockout_results.flux_solution, + knockout_results.flux_solution_ratios, + knockout_results.flux_solution_diffs, + knockout_results.genes_with_metabolic_effects, disease_genes_filename=disease_up_file, ) gene_pairs_up.to_csv(os.path.join(output_dir, f"{context}_Gene_Pairs_Inhi_Fratio_UP.txt"), index=False) diff --git a/main/como/merge_xomics.py b/main/como/merge_xomics.py index 9d264fa1..7079d456 100644 --- a/main/como/merge_xomics.py +++ b/main/como/merge_xomics.py @@ -1,22 +1,24 @@ -#!/usr/bin/python3 +from __future__ import annotations import argparse import json import re import sys from collections import Counter +from dataclasses import dataclass from enum import Enum +from pathlib import Path import numpy as np import pandas as pd from fast_bioservices import BioDBNet, Input, Output +from loguru import logger -from como import proteomics_gen, rnaseq_gen, rpy2_api -from como.como_utilities import split_gene_expression_data +from como import proteomics_gen, return_placeholder_data +from como.combine_distributions import _combine_zscores +from como.custom_types import RNASeqPreparationMethod from como.project import Config - -config = Config() -r_file_path = config.code_dir / "rscripts" / "combine_distributions.R" +from como.utils import split_gene_expression_data class _MergedHeaderNames: @@ -40,34 +42,133 @@ class _HighExpressionHeaderNames: SCRNASEQ = f"{_MergedHeaderNames.SCRNASEQ}_high" -# Merge Output -def merge_logical_table(df: pd.DataFrame): +class AdjustmentMethod(Enum): + """Adjustment method for expression requirement based on differences in number of provided data source types.""" + + PROGRESSIVE = "progressive" + REGRESSIVE = "regressive" + FLAT = "flat" + CUSTOM = "custom" + + @classmethod + def from_string(cls, value: str) -> AdjustmentMethod: + """Convert a string to an AdjustmentMethod enum.""" + if value.lower() not in [t.value for t in cls]: + raise ValueError(f"Adjustment method must be one of {cls}; got: {value}") + return cls(value) + + +@dataclass +class _Arguments: + trnaseq_filename: str + mrnaseq_filename: str + scrnaseq_filename: str + proteomics_filename: str + expression_requirement: str | int + adjustment_method: AdjustmentMethod + no_high_confidence: bool + no_na: bool + merge_zfpkm_distribution: bool + keep_transcriptomics_scores: bool + trna_weight: int + mrna_weight: int + scrna_weight: int + proteomics_weight: int + custom_expression_filename: str + + def __post_init__(self): + if self.expression_requirement.isdigit(): + self.expression_requirement = int(self.expression_requirement) + if self.expression_requirement < 1: + logger.warning( + f"Expression requirement should be at least 1, setting to 1 now. Got {self.expression_requirement}" + ) + self.expression_requirement = 1 + elif self.expression_requirement != "default": + raise ValueError( + f"Expression requirement should be an integer or 'default', got {self.expression_requirement}" + ) + + if self.adjustment_method.value.lower() not in ["progressive", "regressive", "flat", "custom"]: + raise ValueError("Adjust method must be either 'progressive', 'regressive', 'flat', or 'custom'") + + +def _load_rnaseq_tests(filename, context_name, prep_method: RNASeqPreparationMethod) -> tuple[str, pd.DataFrame]: + """Load rnaseq results. + + Returns a dictionary of test (context, context, cell, etc ) names and rnaseq expression data """ - Merge the Rows of Logical Table belongs to the same ENTREZ_GENE_ID - :param df: + config = Config() + + def load_dummy_dict(): + df = return_placeholder_data() + return "dummy", df + + if not filename or filename == "None": # not using this data type, use empty dummy data matrix + return load_dummy_dict() + + inquiry_full_path = config.data_dir / "config_sheets" / filename + if not inquiry_full_path.exists(): # check that config file exist (isn't needed to load, but helps user) + raise FileNotFoundError(f"Error: Config file not found at {inquiry_full_path}") + + match prep_method: + case RNASeqPreparationMethod.TOTAL: + filename = f"rnaseq_total_{context_name}.csv" + case RNASeqPreparationMethod.MRNA: + filename = f"rnaseq_mrna_{context_name}.csv" + case RNASeqPreparationMethod.SCRNA: + filename = f"rnaseq_scrna_{context_name}.csv" + case _: + raise ValueError( + f"Unsupported RNA-seq library type: {prep_method.value}. " + f"Must be an option defined in 'RNASeqPreparationMethod'." + ) + + save_filepath = config.result_dir / context_name / prep_method.value / filename + if save_filepath.exists(): + data = pd.read_csv(save_filepath, index_col="entrez_gene_id") + return context_name, data + + else: + logger.warning( + f"'{prep_method.value}' gene expression file for '{context_name}' was not found at '{save_filepath}'. " + f"If this is not intentional, please fix the filename to match '{save_filepath}'." + ) + return load_dummy_dict() + + +# Merge Output +def _merge_logical_table(df: pd.DataFrame): + """Merge rows of Logical Table belonging to the same entrez_gene_id. + + :param df: pandas dataframe of logical table :return: pandas dataframe of merged table """ # step 1: get all plural ENTREZ_GENE_IDs in the input table, extract unique IDs df.reset_index(drop=False, inplace=True) - df.dropna(axis=0, subset=["ENTREZ_GENE_ID"], inplace=True) - df["ENTREZ_GENE_ID"] = df["ENTREZ_GENE_ID"].str.replace(" /// ", "//").astype(str) + df.dropna(axis=0, subset=["entrez_gene_id"], inplace=True) + df["entrez_gene_id"] = df["entrez_gene_id"].astype(str).str.replace(" /// ", "//").astype(str) + + id_list: list[str] = df.loc[ + ~df["entrez_gene_id"].str.contains("//"), "entrez_gene_id" + ].tolist() # Collect "single" ids, like "123" + multiple_entrez_ids: list[str] = df.loc[ + df["entrez_gene_id"].str.contains("//"), "entrez_gene_id" + ].tolist() # Collect "double" ids, like "123//456" - single_entrez_ids: list[str] = df[~df["ENTREZ_GENE_ID"].str.contains("//")]["ENTREZ_GENE_ID"].tolist() - multiple_entrez_ids: list[str] = df[df["ENTREZ_GENE_ID"].str.contains("//")]["ENTREZ_GENE_ID"].tolist() - id_list: list[str] = [] for i in multiple_entrez_ids: ids = i.split("//") id_list.extend(ids) duplicate_rows = pd.DataFrame([]) for j in ids: - rows = df.loc[df["ENTREZ_GENE_ID"] == i].copy() - rows["ENTREZ_GENE_ID"] = j + rows = df.loc[df["entrez_gene_id"] == i].copy() + rows["entrez_gene_id"] = j duplicate_rows = pd.concat([duplicate_rows, rows], axis=0) df = pd.concat([df, pd.DataFrame(duplicate_rows)], axis=0, ignore_index=True) - df.drop(df[df["ENTREZ_GENE_ID"] == i].index, inplace=True) + df.drop(df[df["entrez_gene_id"] == i].index, inplace=True) full_entrez_id_sets: set[str] = set() entrez_dups_list: list[list[str]] = [] @@ -92,14 +193,13 @@ def merge_logical_table(df: pd.DataFrame): new_entrez_id = " /// ".join(sortlist) full_entrez_id_sets.add(new_entrez_id) - for i in full_entrez_id_sets: - entrez_dups_list.append(i.split(" /// ")) + entrez_dups_list.extend(i.split(" /// ") for i in full_entrez_id_sets) entrez_dups_dict = dict(zip(full_entrez_id_sets, entrez_dups_list)) for merged_entrez_id, entrez_dups_list in entrez_dups_dict.items(): - df["ENTREZ_GENE_ID"].replace(to_replace=entrez_dups_list, value=merged_entrez_id, inplace=True) + df["entrez_gene_id"].replace(to_replace=entrez_dups_list, value=merged_entrez_id, inplace=True) - df.set_index("ENTREZ_GENE_ID", inplace=True) + df.set_index("entrez_gene_id", inplace=True) df = df.fillna(-1).groupby(level=0).max() df.replace(-1, np.nan, inplace=True) @@ -117,28 +217,20 @@ def merge_logical_table(df: pd.DataFrame): return df -def get_transcriptmoic_details(merged_df: pd.DataFrame) -> pd.DataFrame: - """ +async def _get_transcriptmoic_details(merged_df: pd.DataFrame) -> pd.DataFrame: + """Get details of transcriptomic data. + This function will get the following details of transcriptomic data: - Gene Symbol - Gene Name - - ENTREZ_GENE_ID + - entrez_gene_id The resulting dataframe will have its columns created in the order listed above It will return a pandas dataframe with this information - :param merged_df: A dataframe containing all the transcriptomic and proteomic data after determining which genes are active + :param merged_df: A dataframe containing all active transcriptomic and proteomic genes :return: A dataframe with the above-listed columns """ - # import numpy as np - # merged_df["prote_exp"] = np.random.choice([0, 1], size=len(merged_df)) - # merged_df["prote_high"] = pd.NA - # for row in merged_df.itertuples(): - # if row.prote_exp == 1: - # merged_df.at[row.Index, "TotalExpressed"] += 1 - - # merged_df["TotalExpressed"] += 1 if merged_df["prote_exp"] == 1 else 0 - # If _ExpressedHeaderNames.PROTEOMICS.value is in the dataframe, lower the required expression by 1 # We are only trying to get details for transcriptomic data if _ExpressedHeaderNames.PROTEOMICS in merged_df.columns: @@ -153,7 +245,6 @@ def get_transcriptmoic_details(merged_df: pd.DataFrame) -> pd.DataFrame: if required_expression > 1: required_expression -= 1 - # Create a new dataframe without [_ExpressedHeaderNames.PROTEOMICS.value, _HighExpressionHeaderNames.PROTEOMICS.value] columns transcriptomic_df: pd.DataFrame = merged_df.drop( columns=[ _ExpressedHeaderNames.PROTEOMICS, @@ -173,8 +264,8 @@ def get_transcriptmoic_details(merged_df: pd.DataFrame) -> pd.DataFrame: transcriptomic_df: pd.DataFrame = merged_df.copy() biodbnet = BioDBNet() - gene_details: pd.DataFrame = biodbnet.db2db( - input_values=transcriptomic_df.index.astype(str).values.tolist(), + gene_details: pd.DataFrame = await biodbnet.async_db2db( + values=transcriptomic_df.index.astype(str).values.tolist(), input_db=Input.GENE_ID, output_db=[ Output.GENE_SYMBOL, @@ -194,7 +285,8 @@ def get_transcriptmoic_details(merged_df: pd.DataFrame) -> pd.DataFrame: ] gene_details["gene_info_type"] = [ - i.group(1) if isinstance(i, re.Match) else "None" for i in gene_details["Gene Info"].apply(lambda x: re.search(r"\[Gene Type: (.*)\]", x)) + i.group(1) if isinstance(i, re.Match) else "None" + for i in gene_details["Gene Info"].apply(lambda x: re.search(r"\[Gene Type: (.*)\]", x)) ] gene_details["ensembl_info_type"] = [ i.group(1) if isinstance(i, re.Match) else "None" @@ -227,7 +319,7 @@ def get_transcriptmoic_details(merged_df: pd.DataFrame) -> pd.DataFrame: return gene_details -def _merge_xomics( +async def _merge_xomics( context_name: str, expression_requirement, proteomics_file=None, @@ -237,9 +329,7 @@ def _merge_xomics( no_hc=False, no_na=False, ): - """ - Merges rnaseq and/or proteomics active gene logicals from outputs of their respective "_gen.py" - scripts. + """Merge rnaseq and/or proteomics active genes. :param proteomics_file: filename of proteomics config file in /main/data/config_sheets/ :param trnaseq_file: filename of Total RNA-seq config file in /main/data/config_sheets/ @@ -252,15 +342,19 @@ def _merge_xomics( :return: dictionary where keys are contexts, (tissue name, control type etc) and values are expression tables """ config = Config() - print(f"Merging data for {context_name}") + logger.info(f"Merging data for {context_name}") # load data for each source if it exists. IF not load an empty dummy dataset - trnaseq = rnaseq_gen.load_rnaseq_tests(filename=trnaseq_file, context_name=context_name, lib_type="total") - mrnaseq = rnaseq_gen.load_rnaseq_tests(filename=mrnaseq_file, context_name=context_name, lib_type="mrna") - scrnaseq = rnaseq_gen.load_rnaseq_tests(filename=scrnaseq_file, context_name=context_name, lib_type="scrna") + trnaseq = _load_rnaseq_tests( + filename=trnaseq_file, context_name=context_name, prep_method=RNASeqPreparationMethod.TOTAL + ) + mrnaseq = _load_rnaseq_tests( + filename=mrnaseq_file, context_name=context_name, prep_method=RNASeqPreparationMethod.MRNA + ) + scrnaseq = _load_rnaseq_tests( + filename=scrnaseq_file, context_name=context_name, prep_method=RNASeqPreparationMethod.SCRNA + ) proteomics = proteomics_gen.load_proteomics_tests(filename=proteomics_file, context_name=context_name) - files_dict = dict() - expression_list = [] high_confidence_list = [] merge_data = None @@ -317,7 +411,12 @@ def _merge_xomics( ) merge_data = prote_data if merge_data is None else merge_data.join(prote_data, how="outer") - merge_data = merge_logical_table(merge_data) + if merge_data is None: + logger.critical( + f"No data is available for the '{context_name}' context. If this is intentional, ignore this error." + ) + return {} + merge_data = _merge_logical_table(merge_data) num_sources = len(expression_list) merge_data["Active"] = 0 @@ -330,7 +429,9 @@ def _merge_xomics( ) else: # subtract one from requirement per NA merge_data.loc[:, "Required"] = merge_data[expression_list].apply( - lambda x: expression_requirement - (num_sources - x.count()) if (expression_requirement - (num_sources - x.count()) > 0) else 1, + lambda x: expression_requirement - (num_sources - x.count()) + if (expression_requirement - (num_sources - x.count()) > 0) + else 1, axis=1, ) @@ -345,128 +446,124 @@ def _merge_xomics( merge_data = merge_data filepath = config.result_dir / context_name / f"merged_{context_name}.csv" - merge_data.to_csv(filepath, index_label="ENTREZ_GENE_ID") + merge_data.to_csv(filepath, index_label="entrez_gene_id") filepath = config.result_dir / context_name / f"ActiveGenes_{context_name}_Merged.csv" merge_data.reset_index(drop=False, inplace=True) split_entrez = split_gene_expression_data(merge_data) - split_entrez.rename(columns={"Gene": "ENTREZ_GENE_ID", "Data": "Active"}, inplace=True) - split_entrez.to_csv(filepath, index_label="ENTREZ_GENE_ID") - files_dict[context_name] = filepath.as_posix() # Must return as string to be able to serialize + split_entrez.to_csv(filepath, index_label="entrez_gene_id") + files_dict = {context_name: filepath.as_posix()} - transcriptomic_details = get_transcriptmoic_details(merge_data) + transcriptomic_details = await _get_transcriptmoic_details(merge_data) transcriptomic_details_filepath = filepath.parent / f"TranscriptomicDetails_{context_name}.csv" transcriptomic_details.to_csv(transcriptomic_details_filepath, index=False) - print(f"{context_name}: Save to {filepath}\n") + logger.success(f"{context_name}: Save to {filepath}\n") return files_dict -def handle_context_batch( - trnaseq_file, - mrnaseq_file, - scrnaseq_file, - proteomics_file, - tweight, - mweight, - sweight, - pweight, - expression_requirement, - adjust_method, - no_hc, - no_na, - custom_df, - merge_distro, - keep_gene_score, +async def _handle_context_batch( # noqa: C901 + trnaseq_file: Path | None, + mrnaseq_file: Path | None, + scrnaseq_file: Path | None, + proteomics_file: Path | None, + tweight: float, + mweight: float, + sweight: float, + pweight: float, + expression_requirement: int, + adjust_method: AdjustmentMethod, + no_hc: bool, + no_na: bool, + merge_zfpkm_distribution: bool, + keep_gene_score: bool, ): - """ - Handle merging of different data sources for each context type - """ + """Merge different data sources for each context type.""" + if all(file is None for file in [trnaseq_file, mrnaseq_file, scrnaseq_file, proteomics_file]): + raise ValueError("No configuration file was passed!") + config = Config() sheet_names = [] - for file in [ - trnaseq_file, - mrnaseq_file, - scrnaseq_file, - proteomics_file, - ]: + for file in [trnaseq_file, mrnaseq_file, scrnaseq_file, proteomics_file]: if file is not None: config_filepath = config.config_dir / file - xl = pd.ExcelFile(config_filepath, engine="openpyxl") + try: + xl = pd.ExcelFile(config_filepath, engine="openpyxl") + except Exception as e: + raise ValueError(f"Unable to read file '{config_filepath}'") from e sheet_names += xl.sheet_names - use_trna = True if trnaseq_file is not None else False - use_mrna = True if mrnaseq_file is not None else False - use_scrna = True if scrnaseq_file is not None else False - use_proteins = True if proteomics_file is not None else False + use_trna = trnaseq_file is not None + use_mrna = mrnaseq_file is not None + use_scrna = scrnaseq_file is not None + use_proteins = proteomics_file is not None counts = Counter(sheet_names) - sheet_names = sorted(list(set(sheet_names))) - print("The data provided for each context listed will be merged. Data BETWEEN contexts will not be merged, only WITHIN a context") - for i in sheet_names: - # Print the sheet names in a list, like so - # name1, name2, and name3 - print(i, end="") - if counts[i] > 1: - print(f" ({counts[i]}x)", end="") - print(", ", end="") - print("\b\b") - + sheet_names = sorted(set(sheet_names)) + logger.info("Beginning to merge data within contexts") dict_list = {} max_inputs = max(counts.values()) min_inputs = min(counts.values()) - if merge_distro: - print(f"Using {merge_distro} distribution for merging") - rpy2_api.Rpy2( - r_file_path, - config.result_dir.as_posix(), - sheet_names, - use_mrna, - use_trna, - use_scrna, - use_proteins, - keep_gene_score, - tweight, - mweight, - sweight, - pweight, - ).call_function("combine_zscores_main") + if merge_zfpkm_distribution: + logger.debug("Using zFPKM distribution for merging") + _combine_zscores( + working_dir=config.result_dir.as_posix(), + context_names=sheet_names, + global_use_mrna=use_mrna, + global_use_trna=use_trna, + global_use_scrna=use_scrna, + global_use_proteins=use_proteins, + keep_gene_scores=keep_gene_score, + global_trna_weight=tweight, + global_mrna_weight=mweight, + global_scrna_weight=sweight, + global_protein_weight=pweight, + ) for context_name in sheet_names: num_sources = counts[context_name] - if adjust_method == "progressive": - exp_req = (num_sources - min_inputs) + expression_requirement - elif adjust_method == "regressive": - exp_req = expression_requirement - (max_inputs - num_sources) - elif adjust_method == "flat": - exp_req = expression_requirement - else: - exp_req = int(custom_df.iloc[custom_df["context"] == context_name, "req"].iloc[0]) - - print(f"Expression requirement of {expression_requirement} adjusted to {exp_req} using {adjust_method} adjustment method for {context_name}.") + match adjust_method: + case AdjustmentMethod.PROGRESSIVE: + adjusted_expression_requirement = (num_sources - min_inputs) + expression_requirement + case AdjustmentMethod.REGRESSIVE: + adjusted_expression_requirement = expression_requirement - (max_inputs - num_sources) + case AdjustmentMethod.FLAT: + adjusted_expression_requirement = expression_requirement + case _: + adjusted_expression_requirement = int( + custom_df.iloc[custom_df["context"] == context_name, "req"].iloc[0] + ) + + if adjusted_expression_requirement != expression_requirement: + logger.debug( + f"Expression requirement of '{expression_requirement}' adjusted to " + f"'{adjusted_expression_requirement}' using '{adjust_method.value}' adjustment method " + f"for '{context_name}'." + ) - if exp_req > num_sources: - print( - f"WARNING: Expression requirement for {context_name} was calculated to be greater than max number of input data sources." + if adjusted_expression_requirement > num_sources: + logger.warning( + f"Expression requirement for {context_name} was calculated to be greater " + f"than max number of input data sources. " f"Will be force changed to {num_sources} to prevent output from having 0 active genes. " f"Consider lowering the expression requirement or changing the adjustment method." ) - exp_req = num_sources + adjusted_expression_requirement = num_sources - if exp_req < 1: # never allow expression requirement to be less than one - print( - "WARNING: Expression requirement for {context_name} was calculated to be less than 1. " - "Will be force changed to 1 to prevent output from having 0 active genes. " + if adjusted_expression_requirement < 1: # never allow expression requirement to be less than one + logger.warning( + f"Expression requirement for {context_name} was calculated to be less than 1. " + "Will be changed to 1 to prevent output from having 0 active genes. " ) - exp_req = 1 + adjusted_expression_requirement = 1 - files_dict = _merge_xomics( + files_dict = await _merge_xomics( context_name, - expression_requirement=exp_req, + expression_requirement=adjusted_expression_requirement, proteomics_file=proteomics_file, trnaseq_file=trnaseq_file, mrnaseq_file=mrnaseq_file, @@ -475,88 +572,72 @@ def handle_context_batch( no_na=no_na, ) - dict_list.update(files_dict) + dict_list |= files_dict files_json = config.result_dir / "step1_results_files.json" files_json.parent.mkdir(parents=True, exist_ok=True) - with open(files_json.as_posix(), "w") as fp: - json.dump(dict_list, fp) + with files_json.open("w") as f: + json.dump(dict_list, f) # type: ignore return -class AdjustMethod(Enum): - PROGRESSIVE = "progressive" - REGRESSIVE = "regressive" - FLAT = "flat" - CUSTOM = "custom" - - -def merge_xomics( - trnaseq_file: str = None, - mrnaseq_file: str = None, - scrnaseq_file: str = None, - proteomics_file: str = None, - tweight: float = 1, - mweight: float = 1, - sweight: float = 1, - pweight: float = 2, - expression_requirement: int = None, - adjust_method: AdjustMethod = AdjustMethod.FLAT, - no_hc: bool = False, +async def merge_xomics( + trnaseq_filepath: str | Path | None = None, + mrnaseq_filepath: str | Path | None = None, + scrnaseq_filepath: str | Path | None = None, + proteomics_filepath: str | Path | None = None, + trna_weight: float = 1, + mrna_weight: float = 1, + scrna_weight: float = 1, + proteomics_weight: float = 2, + expression_requirement: int | None = None, + adjust_method: AdjustmentMethod = AdjustmentMethod.FLAT, + no_high_confidence: bool = False, no_na: bool = False, - custom_file: str = None, - merge_distro: bool = False, - keep_gene_score: bool = True, + merge_zfpkm_distribution: bool = False, + keep_transcriptomics_score: bool = True, ): - config = Config() - # read custom expression requirment file if used - if custom_file is not None: - custom_filepath = config.data_dir / custom_file - custom_df = pd.read_excel(custom_filepath, sheet_name=0) - custom_df.columns = ["context", "req"] - else: - custom_df = pd.DataFrame([]) - + """Merge expression tables of multiple sources (RNA-seq, proteomics) into one.""" if expression_requirement is None: expression_requirement = sum( - 1 + test is not None for test in [ - trnaseq_file, - mrnaseq_file, - scrnaseq_file, - proteomics_file, + trnaseq_filepath, + mrnaseq_filepath, + scrnaseq_filepath, + proteomics_filepath, ] - if test is not None ) elif expression_requirement < 1: raise ValueError("Expression requirement must be at least 1!") - if adjust_method not in AdjustMethod: - raise ValueError("Adjust method must be either 'progressive', 'regressive', 'flat', or 'custom'") - - handle_context_batch( - trnaseq_file, - mrnaseq_file, - scrnaseq_file, - proteomics_file, - tweight, - mweight, - sweight, - pweight, + trnaseq_filepath = Path(trnaseq_filepath) if trnaseq_filepath else None + mrnaseq_filepath = Path(mrnaseq_filepath) if mrnaseq_filepath else None + scrnaseq_filepath = Path(scrnaseq_filepath) if scrnaseq_filepath else None + proteomics_filepath = Path(proteomics_filepath) if proteomics_filepath else None + + await _handle_context_batch( + trnaseq_filepath, + mrnaseq_filepath, + scrnaseq_filepath, + proteomics_filepath, + trna_weight, + mrna_weight, + scrna_weight, + proteomics_weight, expression_requirement, - adjust_method.value, - no_hc, + adjust_method, + no_high_confidence, no_na, - custom_df, - merge_distro, - keep_gene_score, + merge_zfpkm_distribution, + keep_transcriptomics_score, ) -def main(): - """ - Merge expression tables of multiple sources, (RNA-seq, proteomics) into one list +def _parse_args() -> _Arguments: + """Merge expression tables of multiple sources, (RNA-seq, proteomics) into one list. + User can specify the number of sources with an active gene in order for it to be considered active in the model. Otherwise, it defaults to the number of sources provided. High-confidence genes from any source will be considered active in the model, regardless of agreement with other sources. @@ -570,11 +651,11 @@ def main(): parser.add_argument( "-d", - "--merge-distribution", + "--merge-zfpkm-distribution", action="store_true", required=False, default=False, - dest="merge_distro", + dest="merge_zfpkm_distribution", help="Flag to merge zFPKM distributions. Required if using iMAT reconstruction algorithm in " "create_context_specific_model.py. Must have run rnaseq_gen.py with 'zFPKM' as " "'--technique'. If --proteomics-config-file is given will merge proteomics distributions " @@ -583,11 +664,11 @@ def main(): parser.add_argument( "-k", - "--keep-gene-scores", + "--keep-transcriptomics-scores", action="store_true", required=False, default=True, - dest="keep_gene_score", + dest="keep_transcriptomics_scores", help="When merging z-score distributions of expression, if using both protein abundance and transcipt zFPKM " "flag true if you wish to keep z-score of genes with no protein data, flag false if you wish to discard " "and treat as no expression", @@ -599,7 +680,7 @@ def main(): type=str, required=False, default=None, - dest="trnaseq_file", + dest="trnaseq_filename", help="Name of total RNA-seq config .xlsx file in the /main/data/config_files/.", ) @@ -609,7 +690,7 @@ def main(): type=str, required=False, default=None, - dest="mrnaseq_file", + dest="mrnaseq_filename", help="Name of mRNA-seq config .xlsx file in the /main/data/config_files/.", ) @@ -619,7 +700,7 @@ def main(): type=str, required=False, default=None, - dest="scrnaseq_file", + dest="scrnaseq_filename", help="Name of RNA-seq config .xlsx file in the /main/data/config_files/.", ) @@ -629,7 +710,7 @@ def main(): type=str, required=False, default=None, - dest="proteomics_file", + dest="proteomics_filename", help="Name of proteomics config .xlsx file in the /main/data/config_files/.", ) @@ -640,7 +721,8 @@ def main(): required=False, default=None, dest="expression_requirement", - help="Number of sources with active gene for it to be considered active even if it is not a " "high confidence-gene", + help="Number of sources with active gene for it to be considered active " + "even if it is not a high confidence-gene", ) parser.add_argument( @@ -649,27 +731,29 @@ def main(): type=str, required=False, default="flat", - dest="adjust_method", - help="Technique to adjust expression requirement based on differences in number of provided " "data source types.", + dest="adjustment_method", + help="Technique to adjust expression requirement based on differences in number of provided data source types.", ) parser.add_argument( "-c", "--custom-requirement-file", required="custom" in sys.argv, # required if --requriement-adjust is "custom", - dest="custom_file", - default="SKIP", - help="Name of .xlsx file where first column is context names and second column is expression " "requirement for that context, in /main/data/", + dest="custom_expression_filename", + default=None, + help="Name of .xlsx file where first column is context names and " + "second column is expression requirement for that context", ) parser.add_argument( - "-hc", "--no-hc", + "--no-high-confidence", action="store_true", required=False, default=False, - dest="no_hc", - help="Flag to prevent high-confidence genes forcing a gene to be used in final model " "irrespective of other other data sources", + dest="no_high_confidence", + help="Prevent high-confidence genes forcing a gene to be used in final model, " + "irrespective of other other data sources", ) parser.add_argument( @@ -679,7 +763,7 @@ def main(): required=False, default=False, dest="no_na", - help="Flag to prevent genes missing in a data source library, but present in others from " + help="Prevent genes missing in a data source library, but present in others from " "subtracting 1 from the expression requirement per data source that gene is missing in", ) @@ -689,7 +773,7 @@ def main(): required=False, default=1, type=float, - dest="tweight", + dest="trna_weight", help="Total RNA-seq weight for merging zFPKM distribution", ) @@ -699,7 +783,7 @@ def main(): required=False, default=1, type=float, - dest="mweight", + dest="mrna_weight", help="PolyA enriched (messenger) RNA-seq weight for merging zFPKM distribution", ) @@ -709,7 +793,7 @@ def main(): required=False, default=1, type=float, - dest="sweight", + dest="scrna_weight", help="Single-cell RNA-seq weight for merging zFPKM distribution", ) @@ -719,84 +803,55 @@ def main(): required=False, default=2, type=float, - dest="pweight", + dest="proteomics_weight", help="Proteomics weight for merging z-score distribution", ) - args = parser.parse_args() + args.adjustment_method = AdjustmentMethod.from_string(str(args.adjustment_method)) + return _Arguments(**vars(args)) + + +if __name__ == "__main__": + import asyncio - proteomics_file = args.proteomics_file - trnaseq_file = args.trnaseq_file - mrnaseq_file = args.mrnaseq_file - scrnaseq_file = args.scrnaseq_file - expression_requirement = args.expression_requirement - adjust_method = args.adjust_method.lower() - custom_file = args.custom_file - no_hc = args.no_hc - no_na = args.no_na - merge_distro = args.merge_distro - keep_gene_score = args.keep_gene_score - tweight = args.tweight - mweight = args.mweight - sweight = args.sweight - pweight = args.pweight + args = _parse_args() config = Config() # read custom expression requirment file if used - if custom_file != "SKIP": - custom_filepath = config.data_dir / custom_file + if args.custom_expression_filename: + custom_filepath = config.data_dir / args.custom_expression_filename custom_df = pd.read_excel(custom_filepath, sheet_name=0) custom_df.columns = ["context", "req"] else: custom_df = pd.DataFrame([]) def_exp_req = sum( - [ - 1 - for test in [ - trnaseq_file, - mrnaseq_file, - scrnaseq_file, - proteomics_file, - ] - if test is None + test is None + for test in [ + args.trnaseq_filename, + args.mrnaseq_filename, + args.scrnaseq_filename, + args.proteomics_filename, ] ) - if expression_requirement.lower() == "default": - expression_requirement = def_exp_req - - else: - try: - expression_requirement = int(expression_requirement) - if int(expression_requirement) < 1: - raise ValueError("Expression requirement must be at least 1!") - except ValueError: - raise ValueError("Expression requirement must be able to be converted to an integer!") - - if adjust_method not in ["progressive", "regressive", "flat", "custom"]: - raise ValueError("Adjust method must be either 'progressive', 'regressive', 'flat', or 'custom'") - - handle_context_batch( - trnaseq_file=trnaseq_file, - mrnaseq_file=mrnaseq_file, - scrnaseq_file=scrnaseq_file, - proteomics_file=proteomics_file, - tweight=tweight, - mweight=mweight, - sweight=sweight, - pweight=pweight, - expression_requirement=expression_requirement, - adjust_method=adjust_method, - no_hc=no_hc, - no_na=no_na, - custom_df=custom_df, - merge_distro=merge_distro, - keep_gene_score=keep_gene_score, + asyncio.run( + merge_xomics( + trnaseq_filepath=args.trnaseq_filename, + mrnaseq_filepath=args.mrnaseq_filename, + scrnaseq_filepath=args.scrnaseq_filename, + proteomics_filepath=args.proteomics_filename, + trna_weight=args.trna_weight, + mrna_weight=args.mrna_weight, + scrna_weight=args.scrna_weight, + proteomics_weight=args.proteomics_weight, + expression_requirement=args.expression_requirement, + adjust_method=args.adjustment_method, + no_high_confidence=args.no_high_confidence, + no_na=args.no_na, + merge_zfpkm_distribution=args.merge_zfpkm_distribution, + keep_transcriptomics_score=args.keep_transcriptomics_scores, + ) ) - print("\nDone!") - - -if __name__ == "__main__": - main() + logger.success("Finished merging!") diff --git a/main/como/migrations.py b/main/como/migrations.py new file mode 100644 index 00000000..61ac6abf --- /dev/null +++ b/main/como/migrations.py @@ -0,0 +1,6 @@ +import pandas as pd + + +def gene_info_migrations(df: pd.DataFrame) -> pd.DataFrame: + """Migrate gene info DataFrame to the latest version.""" + return df.rename(columns={"hgnc_symbol": "gene_symbol"}) if "hgnc_symbol" in df.columns else df diff --git a/main/como/project.py b/main/como/project.py index 2a252be2..b6a21bf9 100644 --- a/main/como/project.py +++ b/main/como/project.py @@ -1,14 +1,16 @@ +from __future__ import annotations + from pathlib import Path +from typing import ClassVar + +from loguru import logger class SingletonMeta(type): - _instances = {} + _instances: ClassVar[dict] = {} def __call__(cls, *args, **kwargs): - """ - Possible changes to the value of the `__init__` argument do not affect - the returned instance. - """ + """Validate that changes to the `__init__` argument do not affect the returned instance.""" if cls not in cls._instances: instance = super().__call__(*args, **kwargs) cls._instances[cls] = instance @@ -16,22 +18,64 @@ def __call__(cls, *args, **kwargs): class Config(metaclass=SingletonMeta): - def __init__(self, data_dir: Path = None, config_dir: Path = None, result_dir: Path = None, code_dir: Path = None) -> None: - self.data_dir = data_dir + def __init__( + self, + data_dir: Path | None = None, + config_dir: Path | None = None, + result_dir: Path | None = None, + ) -> None: + """Initialize the Config object.""" + current_dir = Path.cwd() + + self.data_dir = Path(data_dir) if data_dir else None if self.data_dir is None: - self.data_dir = Path.cwd() / "data" - self.data_dir.mkdir(exist_ok=True) + logger.warning(f"'data_dir' not provided to Config, using {Path.cwd() / 'data'}") + self.data_dir = current_dir / "data" + self.data_dir.mkdir(parents=True, exist_ok=True) - self.config_dir = config_dir + self.config_dir = Path(config_dir) if config_dir else None if self.config_dir is None: - self.config_dir = Path.cwd() / "data" / "config_sheets" - self.config_dir.mkdir(exist_ok=True) + logger.warning(f"'config_dir' not provided to Config, using {self.data_dir / 'config_sheets'}") + self.config_dir = self.data_dir / "config_sheets" + self.config_dir.mkdir(parents=True, exist_ok=True) - self.result_dir = result_dir + self.result_dir = Path(result_dir) if result_dir else None if self.result_dir is None: - self.result_dir = Path.cwd() / "data" / "results" - self.result_dir.mkdir(exist_ok=True) + logger.warning(f"'results_dir' not provided to Config, using {self.data_dir / 'results'}") + self.result_dir = self.data_dir / "results" + self.result_dir.mkdir(parents=True, exist_ok=True) + + # Additional directories + self.code_dir = current_dir / "main" / "como" + self.log_dir = self.data_dir / "logs" + self.matrix_dir = self.data_dir / "data_matrices" + self.figures_dir = self.result_dir / "figures" + + self.log_dir.mkdir(parents=True, exist_ok=True) + self.matrix_dir.mkdir(parents=True, exist_ok=True) + self.figures_dir.mkdir(parents=True, exist_ok=True) + + def update(self, **kwargs): + """Update a key in the config object.""" + for key, value in kwargs.items(): + if hasattr(self, key): + setattr(self, key, Path(value) if value else getattr(self, key)) + else: + logger.warning(f"{key} is not a valid attribute of Config") + + def get_context_path(self, context_name: str, create: bool = True) -> Path: + """Get path for a specific context, optionally creating it.""" + path = self.result_dir / context_name + if create: + path.mkdir(parents=True, exist_ok=True) + return path + + def get_r_path(self, path: Path) -> str: + """Convert a Path object to an R-compatible path string.""" + return path.as_posix() - self.code_dir = code_dir - if self.code_dir is None: - self.code_dir = Path.cwd() / "como" + def get_matrix_path(self, context_name: str, filename: str) -> Path: + """Get path for a matrix file in a specific context.""" + path = self.matrix_dir / context_name + path.mkdir(parents=True, exist_ok=True) + return path / filename diff --git a/main/como/proteomics/Crux.py b/main/como/proteomics/Crux.py index 6b5d0a56..b9d0107f 100644 --- a/main/como/proteomics/Crux.py +++ b/main/como/proteomics/Crux.py @@ -1,29 +1,26 @@ -""" -TODO: Integrate crux percolator into this workflow -""" +# ruff: noqa import asyncio - import multiprocessing -from multiprocessing.sharedctypes import Synchronized -import numpy as np import os -import pandas as pd -from pathlib import Path import re import subprocess +from multiprocessing.sharedctypes import Synchronized +from pathlib import Path + +import numpy as np +import pandas as pd import tqdm -from fast_bioservices import BioDBNet, Input, Output, Taxon +from fast_bioservices import BioDBNet + +from como.proteomics.FileInformation import FileInformation, clear_print -from .FileInformation import FileInformation -from .FileInformation import clear_print +# TODO: Integrate crux percolator into this workflow class RAWtoMZML: def __init__(self, file_information: list[FileInformation], core_count: int) -> None: - """ - This class is responsible for converting RAW files to mzML format - """ + """Convert RAW files to mzML format.""" self.file_information: list[FileInformation] = file_information self._core_count: int = core_count @@ -34,11 +31,7 @@ def __init__(self, file_information: list[FileInformation], core_count: int) -> self.raw_to_mzml_wrapper() # Convert from raw files to mzML def raw_to_mzml_wrapper(self) -> None: - """ - This is a wrapper function to multiprocess converting raw files to mzML using ThermoRawFileParser - """ - print("") # Get a new line to print on - + """Multiprocess conversion of raw files to mzML using ThermoRawFileParser.""" # split self.file_information into self._core_count chunks # Always round this division up to ensure all files are processed num_chunks: int = int(np.ceil(len(self.file_information) / self._core_count)) @@ -47,7 +40,7 @@ def raw_to_mzml_wrapper(self) -> None: ] jobs: list[multiprocessing.Process] = [] - for i, information in enumerate(file_chunks): + for information in file_chunks: # Parenthesis + comma needed to make tuple in "args" job = multiprocessing.Process(target=self.raw_to_mzml, args=(information,)) jobs.append(job) @@ -60,9 +53,7 @@ def raw_to_mzml_wrapper(self) -> None: job.terminate() def raw_to_mzml(self, file_information: list[FileInformation]) -> None: - """ - This function is responsible or converting the list of raw files to mzML format - """ + """Convert a list of raw files to mzML format.""" for information in file_information: self._conversion_counter.acquire() self._conversion_counter.value += 1 @@ -72,8 +63,12 @@ def raw_to_mzml(self, file_information: list[FileInformation]) -> None: self._conversion_counter.release() information.mzml_base_path.mkdir(parents=True, exist_ok=True) - subprocess.run( - ["thermorawfileparser", f"--input={str(information.raw_file_path)}", f"--output_file={str(information.mzml_file_path)}"], + subprocess.run( # noqa: S603 + [ # noqa: S607 + "thermorawfileparser", + f"--input={information.raw_file_path!s}", + f"--output_file={information.mzml_file_path!s}", + ], stdout=subprocess.PIPE, ) @@ -85,8 +80,7 @@ def __init__( fasta_database: Path, core_count: int, ) -> None: - """ - This file is responsible for calling the crux-toolkit utilities to process raw proteomics data + """This file is responsible for calling the crux-toolkit utilities to process raw proteomics data The tools here are called through the command line The following steps must be performed: @@ -103,8 +97,7 @@ def __init__( self.mzml_to_sqt() # Analyze mzML files, creating SQT files def mzml_to_sqt(self) -> None: - """ - This function analyzes the converted mzML files and creates SQT files + """This function analyzes the converted mzML files and creates SQT files This function does not use multiprocessing, as Crux Comet incorporates its own multiprocessing """ for i, file_information in enumerate(self._file_information): @@ -135,7 +128,9 @@ def mzml_to_sqt(self) -> None: ) # Replace all "comet.*" in output directory with the name of the file being processed - comet_files = [str(file) for file in os.listdir(file_information.sqt_base_path) if str(file).startswith("comet.")] + comet_files = [ + str(file) for file in os.listdir(file_information.sqt_base_path) if str(file).startswith("comet.") + ] for file_name in comet_files: # Determine the old file path old_file_path: Path = Path(file_information.sqt_base_path, file_name) @@ -148,14 +143,12 @@ def mzml_to_sqt(self) -> None: os.rename(old_file_path, new_file_path) clear_print("SQT creation finished") - print("") + print() class SQTtoCSV: def __init__(self, file_information: list[FileInformation], core_count: int) -> None: - """ - This class is meant to convert UniProt IDs to Entrez IDs using BioDBNet - """ + """This class is meant to convert UniProt IDs to Entrez IDs using BioDBNet""" self._biodbnet: BioDBNet = BioDBNet() self._file_information: list[FileInformation] = file_information self._core_count: int = min(core_count, 4) # Maximum of 4 cores @@ -175,14 +168,11 @@ def __init__(self, file_information: list[FileInformation], core_count: int) -> self.new_write_data() def _uniprot_from_fasta_header(self, fasta_header: str, separator: str = "|") -> str: - """ - This function is responsible for collecting the first-index field from a pipe-separated string - """ + """This function is responsible for collecting the first-index field from a pipe-separated string""" return fasta_header.split(separator)[1] def collect_uniprot_ids_and_ion_intensity(self) -> None: - """ - This function is responsible for collecting the UniProt IDs from the input sqt files + """This function is responsible for collecting the UniProt IDs from the input sqt files Documentation: https://crux.ms/file-formats/sqt-format.html @@ -204,7 +194,7 @@ def collect_uniprot_ids_and_ion_intensity(self) -> None: average_intensities_dict: dict = {key: [] for key in list(file_information.intensity_df.columns)} # average_intensities: pd.DataFrame = pd.DataFrame(columns=["uniprot", replicate_name]) # fmt: skip - with open(file_information.sqt_file_path, "r") as i_stream: + with open(file_information.sqt_file_path) as i_stream: """ We are going to use spectra_line_nums if the list starts with "S" Beneath this, we are going to collect every locus ("L") that does not have a "decoy_" in a nested list @@ -252,17 +242,19 @@ def collect_uniprot_ids_and_ion_intensity(self) -> None: # Assign the file_information intensity dataframe to the gathered values self._file_information[i].intensity_df = pd.DataFrame(average_intensities_dict) - self._file_information[i].intensity_df = self._file_information[i].intensity_df.groupby("uniprot", as_index=False).mean() + self._file_information[i].intensity_df = ( + self._file_information[i].intensity_df.groupby("uniprot", as_index=False).mean() + ) async def _convert_uniprot_wrapper(self) -> None: - """ - This function is a multiprocessing wrapper around the convert_ids function - """ + """This function is a multiprocessing wrapper around the convert_ids function""" values = [self.async_convert_uniprot(self._file_information[i]) for i in range(len(self._file_information))] # Create a progress bar of results # From: https://stackoverflow.com/a/61041328/ - progress_bar = tqdm.tqdm(desc="Starting UniProt to Gene Symbol conversion... ", total=len(self._file_information)) + progress_bar = tqdm.tqdm( + desc="Starting UniProt to Gene Symbol conversion... ", total=len(self._file_information) + ) for i, result in enumerate(asyncio.as_completed(values)): await result # Get result from asyncio.as_completed progress_bar.set_description(f"Working on {i + 1} of {len(self._file_information)}") @@ -306,8 +298,7 @@ async def async_convert_uniprot(self, file_information: FileInformation) -> None file_information.intensity_df.update(gene_symbols) def create_merged_frame(self) -> None: - """ - This function is responsible for merging all dataframes of a specific cell type into a single master frame + """This function is responsible for merging all dataframes of a specific cell type into a single master frame This will allow for aggregating S1R1/S1R2/etc. dataframes into a single S1 dataframe """ for file in self._file_information: @@ -331,8 +322,7 @@ def create_merged_frame(self) -> None: self._merged_frames[cell_type].fillna(0, inplace=True) def split_abundance_values(self) -> None: - """ - This function is responsible for splitting abundance values into separate columns based on their S#R# identifier + """This function is responsible for splitting abundance values into separate columns based on their S#R# identifier It will start by finding all S1R*, S2R*, etc. columns in each cell type under self._master_frames From here, it will merge these dataframes into a new dataframe under self._split_frames, corresponding to the cell type @@ -354,7 +344,7 @@ def split_abundance_values(self) -> None: C ,125 ,175 """ # Get a new line to print output on - print("") + print() # Copy the dictionary keys from merged_frames into the split_frames for key in self._merged_frames.keys(): @@ -388,7 +378,9 @@ def split_abundance_values(self) -> None: # Create a new dataframe to split the S# columns from split_frame: pd.DataFrame = dataframe.copy() # Get the current S{i} columns in - abundance_columns: list[str] = [column for column in split_frame.columns if re.match(rf"{cell_type}_S{i}R\d+", column)] + abundance_columns: list[str] = [ + column for column in split_frame.columns if re.match(rf"{cell_type}_S{i}R\d+", column) + ] take_columns: list[str] = ["symbol"] + abundance_columns average_intensity_name: str = f"{cell_type}_S{i}" @@ -413,9 +405,7 @@ def split_abundance_values(self) -> None: self._split_frames[cell_type].append(split_frame) def new_write_data(self) -> None: - """ - This function is responsible for writing the dataframes found in self._merge_frames to the respective cell type file - """ + """This function is responsible for writing the dataframes found in self._merge_frames to the respective cell type file""" # Get a list of CSV file locations csv_file_location: dict[str, Path] = {} for information in self._file_information: @@ -438,8 +428,7 @@ def new_write_data(self) -> None: self._merged_frames[key].to_csv(csv_file_location[key], index=False) def write_data(self) -> None: - """ - This function creates a unique dataframe for each cell type found in the intensity dataframes + """This function creates a unique dataframe for each cell type found in the intensity dataframes from the self._file_information list It merges these intensity dataframes, creating a new column for each dataframe within each cell type @@ -485,9 +474,8 @@ def write_data(self) -> None: @property def file_information(self) -> list[FileInformation]: - """ - Reassigning the values from the incoming file_manager into a shared-memory variable means - we must provide an option to return the file_information list + """Reassigning the values from the incoming file_manager into a shared-memory variable means + we must provide an option to return the file_information list """ return self._file_information diff --git a/main/como/proteomics/FTPManager.py b/main/como/proteomics/FTPManager.py index 39f94310..1be45d91 100644 --- a/main/como/proteomics/FTPManager.py +++ b/main/como/proteomics/FTPManager.py @@ -1,31 +1,27 @@ -""" -This file is responsible downloading data found at FTP links +# ruff: noqa + +"""This file is responsible downloading data found at FTP links TODO: Find a way to mark a file as "downloaded" - Keep a list of file names in a ".completd" hidden folder? """ -from .FileInformation import FileInformation -from .FileInformation import clear_print -import aioftp import asyncio import multiprocessing -from multiprocessing.sharedctypes import Synchronized import time import typing +from multiprocessing.sharedctypes import Synchronized from urllib.parse import urlparse +import aioftp + +from .FileInformation import FileInformation, clear_print + async def aioftp_client( - host: str, - username: str = "anonymous", - password: str = "guest", - port: int = 21, - max_attempts: int = 3 + host: str, username: str = "anonymous", password: str = "guest", port: int = 21, max_attempts: int = 3 ) -> aioftp.Client: - """ - This class is responsible for creating a "client" connection - """ + """This class is responsible for creating a "client" connection""" connection_successful: bool = False attempt_num: int = 1 @@ -37,17 +33,16 @@ async def aioftp_client( await client.login(user=username, password=password) connection_successful = True except ConnectionResetError: - # Make sure this print statement is on a new line on the first error if attempt_num == 1: - print("") + print() # Line clean: https://stackoverflow.com/a/5419488/13885200 clear_print(f"Attempt {attempt_num} of {max_attempts} failed to connect") attempt_num += 1 time.sleep(5) if not connection_successful: - print("") + print() raise ConnectionResetError("Could not connect to FTP server") return client @@ -55,17 +50,15 @@ async def aioftp_client( class Reader: def __init__( - self, - root_link: str, - file_extensions: list[str], - max_attempts: int = 3, - port: int = 21, - user: str = "anonymous", - passwd: str = "guest" + self, + root_link: str, + file_extensions: list[str], + max_attempts: int = 3, + port: int = 21, + user: str = "anonymous", + passwd: str = "guest", ) -> None: - """ - This class is responsible for reading data about root FTP links - """ + """This class is responsible for reading data about root FTP links""" self._root_link: str = root_link self._extensions: list[str] = file_extensions self._max_attempts: int = max_attempts @@ -75,7 +68,7 @@ def __init__( self._files: list[str] = [] self._file_sizes: list[int] = [] - + self._get_info_wrapper() def _get_info_wrapper(self) -> None: @@ -87,11 +80,9 @@ def _get_info_wrapper(self) -> None: event_loop.close() async def _get_info(self) -> None: - """ - This function is responsible for getting all files under the root_link - """ + """This function is responsible for getting all files under the root_link""" url_parse = urlparse(self._root_link) - + scheme: str host: str folder: str @@ -109,7 +100,7 @@ async def _get_info(self) -> None: raise ValueError(f"Unable to identify folder or path from url: {self._root_link}") client = await aioftp_client(host=host) - for path, info in (await client.list(folder, recursive=True)): + for path, info in await client.list(folder, recursive=True): if str(path).endswith(tuple(self._extensions)): download_url: str = f"{scheme}://{host}{path}" self._files.append(download_url) @@ -140,9 +131,7 @@ def __init__( file_information: list[FileInformation], core_count: int = 1, ) -> None: - """ - This function is responsible for downloading items from the FTP urls passed into it - """ + """This function is responsible for downloading items from the FTP urls passed into it""" self._file_information: list[FileInformation] = file_information self._core_count: int = min(core_count, 2) # Limit to 2 downloads at a time self._download_counter: Synchronized = Synchronized(multiprocessing.Value("i", 1)) @@ -150,11 +139,9 @@ def __init__( # Find files to download self._download_data_wrapper() - + def _download_data_wrapper(self) -> None: - """ - This function is responsible for "kicking off" asynchronous data downloading - """ + """This function is responsible for "kicking off" asynchronous data downloading""" print("Starting file download") event_loop = asyncio.new_event_loop() @@ -172,14 +159,9 @@ def _download_data_wrapper(self) -> None: event_loop.run_until_complete(asyncio.wait(async_tasks)) event_loop.close() - async def _aioftp_download_data( - self, - file_information: FileInformation, - semaphore: asyncio.Semaphore - ) -> None: - + async def _aioftp_download_data(self, file_information: FileInformation, semaphore: asyncio.Semaphore) -> None: url_parse = urlparse(file_information.download_url) - + scheme: str host: str folder: str @@ -197,13 +179,15 @@ async def _aioftp_download_data( raise ValueError(f"Unable to identify folder or path from url: {file_information.download_url}") # Convert file size from byte to MB - size_mb: int = round(file_information.file_size / (1024 ** 2)) + size_mb: int = round(file_information.file_size / (1024**2)) # Use a semaphore so only N number of tasks can be started at once async with semaphore: client = await aioftp_client(host) self._download_counter.acquire() - clear_print(f"Started download {self._download_counter.value:02d} / {len(self._file_information):02d} ({size_mb} MB) - {file_information.raw_file_name}") + clear_print( + f"Started download {self._download_counter.value:02d} / {len(self._file_information):02d} ({size_mb} MB) - {file_information.raw_file_name}" + ) self._download_counter.value += 1 self._download_counter.release() diff --git a/main/como/proteomics/FileInformation.py b/main/como/proteomics/FileInformation.py index e7aca8ad..0b37a4b6 100644 --- a/main/como/proteomics/FileInformation.py +++ b/main/como/proteomics/FileInformation.py @@ -1,23 +1,21 @@ -""" -This will hold all relevant information about a single file to download +# ruff: noqa + +"""This will hold all relevant information about a single file to download This should be implemented as a list of objects """ + import os import sys from pathlib import Path + import pandas as pd -# Add parent directory to path, allows us to import the "project.py" file from the parent directory -# From: https://stackoverflow.com/a/30536516/13885200 -sys.path.append(os.path.dirname(os.path.dirname(os.path.abspath(__file__)))) -import project +from como import project def clear_print(message: str, end: str = "\033[K\r", flush: bool = True) -> None: - """ - Pass in your message exactly as you would like it printed, and this function will clear the screen and print it. - """ + """Pass in your message exactly as you would like it printed, and this function will clear the screen and print it.""" print(message, end=end, flush=flush) @@ -26,7 +24,7 @@ class FileInformation: # This allows us to search through ALL instances of every FileInformation with functions declared here # From: https://stackoverflow.com/a/17253634 instances: list = [] - + def __init__( self, cell_type: str, @@ -42,7 +40,7 @@ def __init__( self.cell_type: str = cell_type self.download_url: str = download_url self.file_size: int = file_size - + # Must check for "None", as we are unable to do study[0] on a None object if isinstance(study, str): self.study: str = study @@ -52,29 +50,30 @@ def __init__( self.study: str = "" self.replicate: str = "" self.batch: str = f"{study}" - + # Base file save paths if raw_path is None: self.raw_base_path: Path = Path(project.configs.data_dir, "results", cell_type, "proteomics", "raw") else: self.raw_base_path: Path = raw_path.parent - + if mzml_path is None: self.mzml_base_path: Path = Path(project.configs.data_dir, "results", cell_type, "proteomics", "mzml") else: self.mzml_base_path: Path = mzml_path.parent - + if sqt_path is None: self.sqt_base_path: Path = Path(project.configs.data_dir, "results", cell_type, "proteomics", "sqt") else: self.sqt_base_path: Path = sqt_path.parent - + if intensity_csv is None: - self.intensity_csv: Path = Path(project.configs.data_dir, "data_matrices", cell_type, - f"protein_abundance_matrix_{cell_type}.csv") + self.intensity_csv: Path = Path( + project.configs.data_dir, "data_matrices", cell_type, f"protein_abundance_matrix_{cell_type}.csv" + ) else: self.intensity_csv: Path = intensity_csv - + # The following variables have inital values set based only on an S# batch, not a replicate # The set_replicate function must be called to set the values for a specific replicate, in which these variables will be reset # File names @@ -82,29 +81,27 @@ def __init__( self.raw_file_name: str = f"{self.base_name}.raw" self.mzml_file_name: str = f"{self.base_name}.mzml" self.sqt_file_name: str = f"{self.base_name}.target.sqt" - + # Full file paths self.raw_file_path: Path = Path(self.raw_base_path, self.raw_file_name) self.mzml_file_path: Path = Path(self.mzml_base_path, self.mzml_file_name) self.sqt_file_path: Path = Path(self.sqt_base_path, self.sqt_file_name) - + # Intensity dataframe self.base_columns: list[str] = ["uniprot"] self.df_columns: list[str] = self.base_columns + [self.batch] self.intensity_df: pd.DataFrame = pd.DataFrame(columns=self.df_columns) - + FileInformation.instances.append(self) - + def set_replicate(self, replicate: str | int): - """ - This function sets self.replicate, and also resets values that use the "replicate" value before it is used - """ + """This function sets self.replicate, and also resets values that use the "replicate" value before it is used""" # Set the initial replicate value if isinstance(replicate, str): self.replicate: str = replicate else: self.replicate: str = f"R{replicate}" - + # "Reset" additional values self.batch: str = f"{self.study}{self.replicate}" # File names @@ -112,29 +109,26 @@ def set_replicate(self, replicate: str | int): self.raw_file_name: str = f"{self.base_name}.raw" self.mzml_file_name: str = f"{self.base_name}.mzml" self.sqt_file_name: str = f"{self.base_name}.target.sqt" - + # Full file paths self.raw_file_path: Path = Path(self.raw_base_path, self.raw_file_name) self.mzml_file_path: Path = Path(self.mzml_base_path, self.mzml_file_name) self.sqt_file_path: Path = Path(self.sqt_base_path, self.sqt_file_name) - + # Intensity dataframe self.base_columns: list[str] = ["uniprot"] self.df_columns: list[str] = self.base_columns + [self.batch] self.intensity_df: pd.DataFrame = pd.DataFrame(columns=self.df_columns) - + @classmethod def filter_instances(cls, cell_type: str): - """ - This function finds all FileInformation objects that have the given cell type - """ + """This function finds all FileInformation objects that have the given cell type""" sorted_instances: list = sorted(cls.instances, key=lambda x: x.study) return [instance for instance in sorted_instances if instance.cell_type == cell_type] - + @staticmethod def intensity_file_path(cell_type: str) -> Path: - """ - This function creates a single instance of the FileInformation class and returns the intensity_csv file location + """This function creates a single instance of the FileInformation class and returns the intensity_csv file location This is useful because each cell type has a specific location all data gets written to If all unique cell types are known, it is then possible to get their intensity csv file location """ diff --git a/main/como/proteomics/__init__.py b/main/como/proteomics/__init__.py index 1c0aa6b2..d78d6d5c 100644 --- a/main/como/proteomics/__init__.py +++ b/main/como/proteomics/__init__.py @@ -1,4 +1 @@ -from . import Crux -from . import FileInformation -from . import FTPManager -from . import proteomics_preprocess +from . import Crux, FileInformation, FTPManager, proteomics_preprocess diff --git a/main/como/proteomics/proteomics_preprocess.py b/main/como/proteomics/proteomics_preprocess.py index ecc67cc3..89264ca2 100644 --- a/main/como/proteomics/proteomics_preprocess.py +++ b/main/como/proteomics/proteomics_preprocess.py @@ -1,52 +1,49 @@ -""" -This is the main driver-file for downloading proteomics data -""" +from __future__ import annotations import argparse import csv import os from pathlib import Path +from loguru import logger + from . import Crux, FTPManager from .FileInformation import FileInformation class ArgParseFormatter(argparse.RawTextHelpFormatter, argparse.ArgumentDefaultsHelpFormatter): - """ - This allows us to use the RawTextHelpFormatter and the ArgumentDefaultsHelpFormatter in a single argparse parser() - """ + """Use the RawTextHelpFormatter and the ArgumentDefaultsHelpFormatter in a single argparse parser().""" pass class ParseCSVInput: - """ - This class is responsible for parsing the input CSV into two fields - 1. proteomeXchange URLs - 2. Cell Type - 3. Replicate (optional) - - This class is meant to make it easier to access each of these things - - - { - "naiveB": { - "url": [one, two, three], - "replicate": [A, B, C], - }, - "nucleophile": { - "url": [four, five, six], - "replicate": [D, E, F], - } - } - """ - def __init__(self, input_csv_file: Path): + """Parse input CSV into two fields. + + 1. proteomeXchange URLs + 2. Cell Type + 3. Replicate (optional) + + This class is meant to make it easier to access each of these things + + + { + "naiveB": { + "url": [one, two, three], + "replicate": [A, B, C], + }, + "nucleophile": { + "url": [four, five, six], + "replicate": [D, E, F], + } + } + """ self._input_csv_file: Path = input_csv_file self._data: [str, dict[str, list[str]]] = {} # Get data from CSV - with open(self._input_csv_file, "r") as i_stream: + with self._input_csv_file.open("w") as i_stream: reader = csv.reader(i_stream) next(reader) for line in reader: @@ -74,13 +71,12 @@ def __init__(self, input_csv_file: Path): @property def ftp_urls(self) -> list[str]: - """ - This will return a list of FTP URLs contained in the input CSV + """Return a list of FTP URLs contained in the input CSV. Example: ftp://ftp.my_server.com """ master_urls: list[str] = [] - for cell_type in self._data.keys(): + for cell_type in self._data: urls = self._data[cell_type]["url"] master_urls.extend(urls) @@ -88,12 +84,12 @@ def ftp_urls(self) -> list[str]: @property def input_cell_types(self) -> list[str]: - """ - This will return the cell types as defined in the input CSV file + """Return the cell types as defined in the input CSV file. + TODO: Match folder paths to correlate S1R1, S1R2, etc.? """ cell_types: list[str] = [] - for key in self._data.keys(): + for key in self._data: # Get the number of URLs per cell type to find the amount of cell types input num_urls: int = len(self._data[key]["url"]) cell_types.extend([key] * num_urls) @@ -101,19 +97,17 @@ def input_cell_types(self) -> list[str]: @property def studies(self) -> list[str]: - """ - This will return the replicates as defined in the input CSV file - """ + """Return the replicates as defined in the input CSV file.""" master_studies: list[str] = [] - for cell_type in self._data.keys(): + for cell_type in self._data: replicates = self._data[cell_type]["study"] master_studies.extend(replicates) return master_studies @property def csv_dict(self) -> dict[str, dict[str, list[str]]]: - """ - This function returns the data dictionary + """Return the CSV information as a dictionary. + It contains data in the following format { CELL_TYPE_1: { @@ -135,8 +129,9 @@ def __init__( file_information: list[FileInformation], csv_data: ParseCSVInput, skip_download: bool, - preferred_extensions: list[str] = None, + preferred_extensions: list[str] | None = None, ): + """Populate FileInformation list with data from the input CSV file.""" self.file_information: list[FileInformation] = file_information self._csv_data: ParseCSVInput = csv_data self._csv_data: dict[str, dict[str, list[str]]] = csv_data.csv_dict @@ -157,54 +152,44 @@ def _gather_data(self): # Iterate through the cell type and corresponding list of URLS # cell_type: naiveB # ftp_urls: ["url_1", "url_2"] - for i, cell_type in enumerate(self._csv_data.keys()): + for cell_type in self._csv_data: ftp_urls: list[str] = self._csv_data[cell_type]["url"] studies: list[str] = self._csv_data[cell_type]["study"] url_count = 0 # Iterate through the URLs available - for j, (url, study) in enumerate(zip(ftp_urls, studies)): - # Print updates to he screen - print( - f"\rParsing cell type {i + 1} of {len(self._csv_data.keys())} ({cell_type}) | {j + 1} of {len(ftp_urls)} folders navigated", - end="", - flush=True, + for url, study in zip(ftp_urls, studies): + ftp_data: FTPManager.Reader = FTPManager.Reader( + root_link=url, file_extensions=self._preferred_extensions ) - ftp_data: FTPManager.Reader = FTPManager.Reader(root_link=url, file_extensions=self._preferred_extensions) - - urls = [url for url in ftp_data.files] - sizes = [size for size in ftp_data.file_sizes] + urls = list(ftp_data.files) + sizes = list(ftp_data.file_sizes) url_count += len(urls) # Iterate through all files and sizes found for url_## - for k, (file, size) in enumerate(zip(urls, sizes)): - self.file_information.append(FileInformation(cell_type=cell_type, download_url=file, file_size=size, study=study)) - - # Print number of URLs found for each cell type - # This is done after appending because some cell types may have more than 1 root URL, and it messes up the formatting - if url_count == 1: - print(" | 1 file found") - else: - print(f" | {url_count} files found") + for file, size in zip(urls, sizes): + self.file_information.append( + FileInformation(cell_type=cell_type, download_url=file, file_size=size, study=study) + ) def print_download_size(self): - # Print the total size to download if we must download data + """Print the total size to download if we must download data.""" total_size: int = 0 for information in self.file_information: total_size += information.file_size # Convert to MB total_size = total_size // 1024**2 - print(f"Total size to download: {total_size} MB") + logger.info(f"Total size to download: {total_size} MB") def _set_replicate_numbers(self): instances: dict[str, list[FileInformation]] = {} for information in self.file_information: - if information.cell_type not in instances.keys(): + if information.cell_type not in instances: instances[information.cell_type] = FileInformation.filter_instances(information.cell_type) - for cell_type in instances.keys(): + for cell_type in instances: replicate_num: int = 1 for i, file_information in enumerate(instances[cell_type]): current_info: FileInformation = file_information @@ -223,22 +208,12 @@ def _set_replicate_numbers(self): current_info.set_replicate(replicate_value) def _collect_cell_type_information(self, cell_type: str) -> list[FileInformation]: - """ - This function is responsible for collecting all FileInformation objects of a given cell type - """ - file_information_list: list[FileInformation] = [] - for information in self.file_information: - if information.cell_type == cell_type: - file_information_list.append(information) - - return file_information_list + """Collect all FileInformation objects of a given cell type.""" + return [information for information in self.file_information if information.cell_type == cell_type] def parse_args() -> argparse.Namespace: - """ - This function is used to parse arguments from the command line - """ - + """Parse arguments from the command line.""" parser = argparse.ArgumentParser( prog="proteomics_preprocess.py", description="Download and analyze proteomics data from proteomeXchange\n" @@ -285,7 +260,7 @@ def parse_args() -> argparse.Namespace: dest="skip_download", default=False, type=bool, - help="If this action is passed in, FTP data will not be downloaded.\n" + help="If this action is passed in, FTP data will not be downloaded. " "This assumes you have raw data under the folder specified by the option '--ftp-out-dir'", ) parser.add_argument( @@ -294,9 +269,9 @@ def parse_args() -> argparse.Namespace: dest="skip_mzml", type=bool, default=False, - help="If this action is passed in, files will not be converted from RAW to mzML format.\n" - "This assumes you have mzML files under the folder specified by the option '--mzml-out-dir'.\n" - "This will continue the workflow from SQT file creation -> CSV ion intensity creation.\n" + help="If this action is passed in, files will not be converted from RAW to mzML format. " + "This assumes you have mzML files under the folder specified by the option '--mzml-out-dir'. " + "This will continue the workflow from SQT file creation -> CSV ion intensity creation. " "If this option is passed in, FTP data will also not be downloaded.", ) parser.add_argument( @@ -305,9 +280,11 @@ def parse_args() -> argparse.Namespace: dest="skip_sqt", type=bool, default=False, - help="If this action is passed in, SQT files will not be created. This assumes you have SQT files under the folder specified by the option '--sqt-out-dir'.\n" - "This will only read data from SQT files and create a CSV ion intensity file.\n" - "If this option is passed in, FTP data will not be downloaded, RAW files will not be converted, and SQT files will not be created.", + help="If this action is passed in, SQT files will not be created. " + "This assumes you have SQT files under the folder specified by the option '--sqt-out-dir'. " + "This will only read data from SQT files and create a CSV ion intensity file. " + "If this option is passed in, FTP data will not be downloaded, " + "RAW files will not be converted, and SQT files will not be created.", ) parser.add_argument( "-c", @@ -316,41 +293,42 @@ def parse_args() -> argparse.Namespace: dest="core_count", metavar="cores", default=os.cpu_count() // 2, - help="This is the number of threads to use for downloading files.\n" - "It will default to the minimum of: half the available CPU cores available, or the number of input files found.\n" - "It will not use more cores than necessary\n" - "Options are an integer or 'all' to use all available cores.\n" - "Note: Downloading will use a MAX of 2 threads at once, as some FTP servers do not work well with multiple connections from the same IP address at once.", + help="This is the number of threads to use for downloading files. " + "It will default to the minimum of: half the available CPU cores available, " + "or the number of input files found. " + "It will not use more cores than necessary. " + "Options are an integer or 'all' to use all available cores. " + "Note: Downloading will use a MAX of 2 threads at once, " + "as some FTP servers do not work well with multiple connections from the same IP address at once.", ) # TODO: Add option to delete intermediate files (raw, mzml, sqt) - args: argparse.Namespace = parser.parse_args(args) + args: argparse.Namespace = parser.parse_args() args.extensions = args.extensions.split(",") # Validte the input file exists if not Path(args.input_csv).is_file(): - print(f"Input file {args.input} does not exist!") - raise FileNotFoundError + raise FileNotFoundError(f"Input file {args.input} does not exist!") if args.core_count == "all": args.core_count = os.cpu_count() - elif not args.core_count.is_integer(): - raise ValueError(f"Invalid option '{args.core_count}' for option '--cores'. Enter an integer or 'all' to use all cores") + elif not str(args.core_count).isdigit(): + raise ValueError( + f"Invalid option '{args.core_count}' for option '--cores'. Enter an integer or 'all' to use all cores" + ) else: args.core_count = int(args.core_count) if args.core_count > os.cpu_count(): - print(f"{args.core_count} cores not available, system only has {os.cpu_count()} cores. Setting '--cores' to {os.cpu_count()}") + logger.info( + f"{args.core_count} cores not available, system only has {os.cpu_count()} cores. " + f"Setting '--cores' to {os.cpu_count()}" + ) args.core_count = os.cpu_count() return args -def main(): - """ - This is the main driver function - - :param args: The list of arguments collected from the command line - """ +def _main(): file_information: list[FileInformation] = [] args: argparse.Namespace = parse_args() csv_data = ParseCSVInput(args.input_csv) @@ -377,7 +355,12 @@ def main(): args.skip_download = True # Populate the file_information list - PopulateInformation(file_information=file_information, csv_data=csv_data, skip_download=args.skip_download, preferred_extensions=args.extensions) + PopulateInformation( + file_information=file_information, + csv_data=csv_data, + skip_download=args.skip_download, + preferred_extensions=args.extensions, + ) # Download data if we should not skip anything if args.skip_download is False: @@ -386,7 +369,6 @@ def main(): file_information=file_information, core_count=args.core_count, ) - print("") # New line to separate this output from the next if args.skip_mzml is False: # Convert raw to mzML and then create SQT files @@ -394,7 +376,6 @@ def main(): file_information=file_information, core_count=args.core_count, ) - print("") # New line to separate this output from the next if args.skip_sqt is False: # Convert mzML to SQT @@ -409,14 +390,7 @@ def main(): file_information=file_information, core_count=args.core_count, ) - print("") # New line to separate this output from the next - - # Get the root folder of output CSV file - root_folders: set[Path] = set([i.intensity_csv.parent for i in file_information]) - print("\nProtein intensities saved under:") - for folder in root_folders: - print(folder) if __name__ == "__main__": - main() + _main() diff --git a/main/como/proteomics_gen.py b/main/como/proteomics_gen.py index 80ff274a..b1e17b34 100644 --- a/main/como/proteomics_gen.py +++ b/main/como/proteomics_gen.py @@ -1,93 +1,81 @@ -#!/usr/bin/python3 - import argparse -import sys -from pathlib import Path +import asyncio import numpy as np import pandas as pd -from fast_bioservices.biodbnet import BioDBNet, Input, Output, Taxon +from fast_bioservices.biodbnet import BioDBNet, Input, Output +from loguru import logger -from como import rpy2_api +from como import return_placeholder_data from como.project import Config - -r_file_path = Path(__file__).parent / "rscripts" / "protein_transform.R" +from como.proteomics_preprocessing import protein_transform_main # Load Proteomics def load_proteomics_data(datafilename, context_name): - """ - Add description...... - """ + """Load proteomics data from a given context and filename.""" config = Config() data_path = config.data_dir / "data_matrices" / context_name / datafilename - print('Data matrix is at "{}"'.format(data_path)) + logger.info(f"Data Matrix Path: {data_path}") if data_path.exists(): proteomics_data = pd.read_csv(data_path, header=0) - else: - print("Error: file not found: {}".format(data_path)) + logger.error(f"Error: file not found: {data_path}") return None # Preprocess data, drop na, duplicate ';' in symbol, - proteomics_data["symbol"] = proteomics_data["symbol"].astype(str) - proteomics_data.dropna(subset=["symbol"], inplace=True) - pluralnames = proteomics_data[proteomics_data["symbol"].str.contains(";") == True] + proteomics_data["gene_symbol"] = proteomics_data["gene_symbol"].astype(str) + proteomics_data.dropna(subset=["gene_symbol"], inplace=True) + pluralnames = proteomics_data[proteomics_data["gene_symbol"].str.contains(";") == True] # noqa: E712 for idx, row in pluralnames.iterrows(): - names = row["symbol"].split(";") + names = row["gene_symbol"].split(";") rows = [] for name in names: rowcopy = row.copy() - rowcopy["symbol"] = name + rowcopy["gene_symbol"] = name rows.append(rowcopy) proteomics_data.drop(index=idx, inplace=True) proteomics_data = pd.concat([proteomics_data, pd.DataFrame(rows)], ignore_index=True) - proteomics_data.rename(columns={"symbol": "Gene Symbol"}, inplace=True) - return proteomics_data # read map to convert to entrez -def load_gene_symbol_map(gene_symbols, filename="proteomics_entrez_map.csv"): - """ - Add descirption.... - """ +async def load_gene_symbol_map(gene_symbols: list[str]): + """Add descirption....""" config = Config() filepath = config.data_dir / "proteomics_entrez_map.csv" if filepath.exists(): - df = pd.read_csv(filepath, index_col="Gene Symbol") + df = pd.read_csv(filepath, index_col="gene_symbol") else: biodbnet = BioDBNet() - df = biodbnet.db2db(input_values=gene_symbols, input_db=Input.GENE_SYMBOL, output_db=[Output.GENE_ID, Output.ENSEMBL_GENE_ID]) - df.loc[df["Gene ID"] == "-", ["Gene ID"]] = np.nan - df.to_csv(filepath, index_label="Gene Symbol") + df = await biodbnet.async_db2db( + values=gene_symbols, input_db=Input.GENE_SYMBOL, output_db=[Output.GENE_ID, Output.ENSEMBL_GENE_ID] + ) + df.loc[df["gene_id"] == "-", ["gene_id"]] = np.nan + df.to_csv(filepath, index_label="gene_symbol") return df[~df.index.duplicated()] def abundance_to_bool_group(context_name, group_name, abundance_matrix, rep_ratio, hi_rep_ratio, quantile): - """ - Descrioption.... - """ + """Descrioption....""" config = Config() output_dir = config.result_dir / context_name / "proteomics" output_dir.mkdir(parents=True, exist_ok=True) # write group abundances to individual files - abundance_filepath = config.result_dir / context_name / "proteomics" / "".join(["protein_abundance_", group_name, ".csv"]) - abundance_matrix.to_csv(abundance_filepath, index_label="ENTREZ_GENE_ID") - - # Z-tranform - rpy2_api.Rpy2(r_file_path, abundance_filepath, output_dir, group_name).call_function("protein_transform_main") - # protein_transform_io.protein_transform_main(abundance_filepath, output_dir, group_name) + abundance_filepath = ( + config.result_dir / context_name / "proteomics" / "".join(["protein_abundance_", group_name, ".csv"]) + ) + abundance_matrix.to_csv(abundance_filepath, index_label="entrez_gene_id") + protein_transform_main(abundance_matrix, output_dir, group_name) # Logical Calculation - thresholds = abundance_matrix.quantile(quantile, axis=0) abundance_matrix_nozero = abundance_matrix.replace(0, np.nan) thresholds = abundance_matrix_nozero.quantile(quantile, axis=0) testbool = pd.DataFrame(0, columns=list(abundance_matrix), index=abundance_matrix.index) @@ -102,20 +90,21 @@ def abundance_to_bool_group(context_name, group_name, abundance_matrix, rep_rati abundance_matrix.loc[(abundance_matrix["pos"] >= hi_rep_ratio), ["high"]] = 1 bool_filepath = output_dir / f"bool_prot_Matrix_{context_name}_{group_name}.csv" - abundance_matrix.to_csv(bool_filepath, index_label="ENTREZ_GENE_ID") + abundance_matrix.to_csv(bool_filepath, index_label="entrez_gene_id") def to_bool_context(context_name, group_ratio, hi_group_ratio, group_names): + """Convert proteomic data to boolean expression.""" config = Config() output_dir = config.result_dir / context_name / "proteomics" - merged_df = pd.DataFrame(columns=["ENTREZ_GENE_ID", "expressed", "high"]) - merged_df.set_index(["ENTREZ_GENE_ID"], inplace=True) + merged_df = pd.DataFrame(columns=["entrez_gene_id", "expressed", "high"]) + merged_df.set_index(["entrez_gene_id"], inplace=True) merged_hi_df = merged_df for group in group_names: read_filepath = output_dir / f"bool_prot_Matrix_{context_name}_{group}.csv" read_df = pd.read_csv(read_filepath) - read_df.set_index("ENTREZ_GENE_ID", inplace=True) + read_df.set_index("entrez_gene_id", inplace=True) read_df = read_df[["expressed", "high"]] if not merged_df.empty: @@ -132,21 +121,17 @@ def to_bool_context(context_name, group_ratio, hi_group_ratio, group_names): out_df = pd.merge(merged_df, merged_hi_df, right_index=True, left_index=True) out_filepath = output_dir / f"Proteomics_{context_name}.csv" - out_df.to_csv(out_filepath, index_label="ENTREZ_GENE_ID") - print("Test Data Saved to {}".format(out_filepath)) + out_df.to_csv(out_filepath, index_label="entrez_gene_id") + logger.success(f"Test Data Saved to {out_filepath}") # read data from csv files def load_proteomics_tests(filename, context_name): - """ - Description.... - """ + """Load statistical test results.""" config = Config() def load_empty_dict(): - savepath = config.data_dir / "data_matrices" / "placeholder" / "placeholder_empty_data.csv" - dat = pd.read_csv(savepath, index_col="ENTREZ_GENE_ID") - return "dummy", dat + return "dummy", return_placeholder_data() if not filename or filename == "None": # if not using proteomics load empty dummy data matrix return load_empty_dict() @@ -155,21 +140,30 @@ def load_empty_dict(): if not inquiry_full_path.exists(): raise FileNotFoundError(f"Error: file not found {inquiry_full_path}") - filename = "Proteomics_{}.csv".format(context_name) + filename = f"Proteomics_{context_name}.csv" full_save_filepath = config.result_dir / context_name / "proteomics" / filename if full_save_filepath.exists(): - data = pd.read_csv(full_save_filepath, index_col="ENTREZ_GENE_ID") - print(f"Read from {full_save_filepath}") + data = pd.read_csv(full_save_filepath, index_col="entrez_gene_id") + logger.success(f"Read from {full_save_filepath}") return context_name, data else: - print(f"Proteomics gene expression file for {context_name} was not found at {full_save_filepath}. Is this intentional?") + logger.warning( + f"Proteomics gene expression file for {context_name} was not found at {full_save_filepath}. " + f"Is this intentional?" + ) return load_empty_dict() -def proteomics_gen( - config_file: str, rep_ratio: float = 0.5, group_ratio: float = 0.5, hi_rep_ratio: float = 0.5, hi_group_ratio: float = 0.5, quantile: int = 25 +async def proteomics_gen( + config_file: str, + rep_ratio: float = 0.5, + group_ratio: float = 0.5, + hi_rep_ratio: float = 0.5, + hi_group_ratio: float = 0.5, + quantile: int = 25, ): + """Generate proteomics data.""" config = Config() if not config_file: raise ValueError("Config file must be provided") @@ -179,7 +173,7 @@ def proteomics_gen( quantile /= 100 prot_config_filepath = config.data_dir / "config_sheets" / config_file - print('Config file is at "{}"'.format(prot_config_filepath)) + logger.info(f"Config file is at '{prot_config_filepath}'") xl = pd.ExcelFile(prot_config_filepath) sheet_names = xl.sheet_names @@ -187,42 +181,31 @@ def proteomics_gen( for context_name in sheet_names: datafilename = "".join(["protein_abundance_", context_name, ".csv"]) config_sheet = pd.read_excel(prot_config_filepath, sheet_name=context_name) - groups = config_sheet["Group"].unique().tolist() + groups = config_sheet["group"].unique().tolist() for group in groups: - group_idx = np.where([True if g == group else False for g in config_sheet["Group"].tolist()]) - cols = np.take(config_sheet["SampleName"].to_numpy(), group_idx).ravel().tolist() + ["Gene Symbol", "uniprot"] - cols = np.take(config_sheet["SampleName"].to_numpy(), group_idx).ravel().tolist() + ["Gene Symbol"] + group_idx = np.where([g == group for g in config_sheet["group"].tolist()]) + cols = [*np.take(config_sheet["sample_name"].to_numpy(), group_idx).ravel().tolist(), "gene_symbol"] proteomics_data = load_proteomics_data(datafilename, context_name) proteomics_data = proteomics_data.loc[:, cols] - sym2id = load_gene_symbol_map(gene_symbols=proteomics_data["Gene Symbol"].tolist(), filename="proteomics_entrez_map.csv") - - # map gene symbol to ENTREZ_GENE_ID - proteomics_data.dropna(subset=["Gene Symbol"], inplace=True) - - try: + symbols_to_ids = await load_gene_symbol_map(gene_symbols=proteomics_data["gene_symbol"].tolist()) + proteomics_data.dropna(subset=["gene_symbol"], inplace=True) + if "uniprot" in proteomics_data.columns: proteomics_data.drop(columns=["uniprot"], inplace=True) - except KeyError: - pass - - proteomics_data = proteomics_data.groupby(["Gene Symbol"]).agg("max") - proteomics_data["ENTREZ_GENE_ID"] = sym2id["Gene ID"] - proteomics_data.dropna(subset=["ENTREZ_GENE_ID"], inplace=True) - proteomics_data.set_index("ENTREZ_GENE_ID", inplace=True) + proteomics_data = proteomics_data.groupby(["gene_symbol"]).agg("max") + proteomics_data["entrez_gene_id"] = symbols_to_ids["gene_id"] + proteomics_data.dropna(subset=["entrez_gene_id"], inplace=True) + proteomics_data.set_index("entrez_gene_id", inplace=True) # save proteomics data by test abundance_to_bool_group(context_name, group, proteomics_data, rep_ratio, hi_rep_ratio, quantile) - to_bool_context(context_name, group_ratio, hi_group_ratio, groups) -def main(argv): - """ - Description - """ +def _main(): parser = argparse.ArgumentParser( prog="proteomics_gen.py", description="Description goes here", @@ -281,21 +264,21 @@ def main(argv): required=False, default=25, dest="quantile", - help="The quantile of genes to accept. This should be an integer from 0% (no proteins pass) " "to 100% (all proteins pass).", + help="The quantile of genes to accept. This should be an integer from 0% (no proteins pass) " + "to 100% (all proteins pass).", ) args = parser.parse_args() - - suppfile = args.config_file - rep_ratio = args.rep_ratio - group_ratio = args.group_ratio - hi_rep_ratio = args.hi_rep_ratio - hi_group_ratio = args.hi_group_ratio - quantile = args.quantile - - proteomics_gen(suppfile, rep_ratio, group_ratio, hi_rep_ratio, hi_group_ratio, quantile) - - return True + asyncio.run( + proteomics_gen( + args.config_file, + args.rep_ratio, + args.group_ratio, + args.hi_rep_ratio, + args.hi_group_ratio, + args.quantile, + ) + ) if __name__ == "__main__": - main(sys.argv[1:]) + _main() diff --git a/main/como/proteomics_preprocessing.py b/main/como/proteomics_preprocessing.py new file mode 100644 index 00000000..9b215439 --- /dev/null +++ b/main/como/proteomics_preprocessing.py @@ -0,0 +1,151 @@ +from __future__ import annotations + +import colorsys +import random +from dataclasses import dataclass +from pathlib import Path + +import numpy as np +import numpy.typing as npt +import pandas as pd +import plotly.graph_objs as go +from loguru import logger +from plotly.subplots import make_subplots +from scipy import stats +from scipy.signal import find_peaks +from sklearn.neighbors import KernelDensity + + +@dataclass +class ZResult: + """Dataclass to store results of Z-score calculation.""" + + zfpkm: pd.DataFrame + x_range: pd.DataFrame # npt.NDArray[np.float64] + density: pd.DataFrame # npt.NDArray[np.float64] + mu: npt.NDArray[np.float64] + std_dev: npt.NDArray[np.float64] + max_fpkm_peak: npt.NDArray[np.float64] + + +def z_score_calc(abundance: pd.DataFrame, min_thresh: int) -> ZResult: + """Calculate Z-scores for protein abundance data.""" + values = abundance.values.copy() + 1 + log_abundance_filt = np.log2(values[values > min_thresh]).reshape((len(abundance), len(abundance.columns))) + log_abundance = np.log2(values) + + # np.zeros((1000, len(abundance.columns)), dtype=np.float64), + z_result = ZResult( + zfpkm=pd.DataFrame( + data=np.nan * np.ones_like(values), index=abundance.index, columns=abundance.columns, dtype=np.float64 + ), + x_range=pd.DataFrame( + data=np.zeros((1000, len(abundance.columns))), columns=abundance.columns, dtype=np.float64 + ), + density=pd.DataFrame( + data=np.zeros((1000, len(abundance.columns))), columns=abundance.columns, dtype=np.float64 + ), + mu=np.zeros(len(abundance.columns)), + std_dev=np.zeros(len(abundance.columns)), + max_fpkm_peak=np.zeros(len(abundance.columns)), + ) + + for i, col in enumerate(abundance.columns): + kde = KernelDensity(kernel="gaussian", bandwidth=0.5).fit(log_abundance_filt[:, i].reshape(-1, 1)) + x_range = np.linspace(log_abundance[:, i].min(), log_abundance[:, i].max(), 1000) + density = np.exp(kde.score_samples(x_range.reshape(-1, 1))) # type: ignore + peaks, _ = find_peaks(density, height=0.02, distance=1.0) + peak_positions = x_range[peaks] + + mu = peak_positions.max() + max_fpkm_peak = density[peaks[np.argmax(peak_positions)]] # type: ignore + + # Select rows from `log_abundance` that are greater than 0 and less than mu in the column `i` + u = log_abundance[:, i][(log_abundance[:, i] > 0) & (log_abundance[:, i] < mu)].mean() + std_dev = (mu - u) * np.sqrt(np.pi / 2) + zfpkm_values = (log_abundance[:, i] - mu) / std_dev + + z_result.zfpkm[col] = zfpkm_values + z_result.x_range[col] = x_range + z_result.density[col] = density + z_result.mu[i] = mu + z_result.std_dev[i] = std_dev + z_result.max_fpkm_peak[i] = max_fpkm_peak + + return z_result + + +def lighten_color(red: int, green: int, blue: int, factor: float = 0.5) -> str: + """Lighten a color by a given factor.""" + # Convert RGB to HLS + hue, lightness, saturation = colorsys.rgb_to_hls(red / 255.0, green / 255.0, blue / 255.0) + + # Increase lightness + lightness = min(1.0, lightness + (1 - lightness) * factor) + + # Covnert back to RGB values + r, g, b = colorsys.hsv_to_rgb(hue, saturation, lightness) + return f"rgb({int(r * 255)},{int(g * 255)},{int(b * 255)})" + + +# Plotting function +def plot_gaussian_fit(z_results: ZResult, facet_titles: bool = True, x_min: int = -4) -> go.Figure: + """Plot Gaussian fit for Z-score transformed data.""" + zfpkm = z_results.zfpkm + x_range = z_results.x_range + density = z_results.density + mu = z_results.mu + std_dev = z_results.std_dev + max_fpkm = z_results.max_fpkm_peak + + fig = make_subplots( + rows=len(zfpkm.columns), cols=1, subplot_titles=zfpkm.columns if facet_titles else [None] * len(zfpkm.columns) + ) + for i, col in enumerate(zfpkm.columns): + fitted = stats.norm.pdf(x_range[col], loc=mu[i], scale=std_dev[i]) + scale_fit = fitted * (max_fpkm[i] / fitted.max()) + + # Select random RGB values for each plot + r = random.randint(0, 255) # noqa: S311, not for cryptographic purposes + g = random.randint(0, 255) # noqa: S311, not for cryptographic purposes + b = random.randint(0, 255) # noqa: S311, not for cryptographic purposes + color, lighten = f"rgb({r},{g},{b})", lighten_color(r, g, b) + fig.add_trace( + go.Scatter(x=x_range[col], y=density[col], name="Abundance Density", line={"color": color}), + row=i + 1, + col=1, + ) + fig.add_trace( + go.Scatter(x=x_range[col], y=scale_fit, name="Fitted Gaussian", line={"dash": "dash", "color": lighten}), + row=i + 1, + col=1, + ) + + fig.update_xaxes(title_text="log2(abundance)", range=[x_min, x_range[col].max()], row=i + 1, col=1) + fig.update_yaxes(title_text="[scaled] density", row=i + 1, col=1) + + fig.update_layout(height=len(zfpkm.columns) * 250, width=800, title_text="Gaussian Fit per Sample") + return fig + + +# Main function for protein abundance transformation +def protein_transform_main(abundance_df: pd.DataFrame | str | Path, out_dir: str | Path, group_name: str) -> None: + """Transform protein abundance data.""" + out_dir: Path = Path(out_dir) + output_figure_directory = out_dir / "figures" + output_figure_directory.mkdir(parents=True, exist_ok=True) + + abundance_df: pd.DataFrame = ( + pd.read_csv(abundance_df) if isinstance(abundance_df, (str, Path)) else abundance_df.fillna(0) + ) + abundance_df = abundance_df[np.isfinite(abundance_df).all(axis=1)] # Remove +/- infinity values + z_transform: ZResult = z_score_calc(abundance_df, min_thresh=0) + + fig = plot_gaussian_fit(z_results=z_transform, facet_titles=True, x_min=-4) + fig.write_image(out_dir / "gaussian_fit.png") + fig.write_html(out_dir / "gaussian_fit.html") + logger.info(f"Wrote image to {out_dir / 'gaussian_fit.png'}") + + z_transformed_abundances = z_transform.zfpkm + z_transformed_abundances[abundance_df == 0] = -4 + z_transformed_abundances.to_csv(out_dir / f"protein_zscore_Matrix_{group_name}.csv", index=False) diff --git a/main/como/rnaseq.py b/main/como/rnaseq.py new file mode 100644 index 00000000..1c504d97 --- /dev/null +++ b/main/como/rnaseq.py @@ -0,0 +1,763 @@ +from __future__ import annotations + +import gc +import math +import multiprocessing +import time +from collections import namedtuple +from dataclasses import dataclass, field +from enum import Enum +from functools import partial +from multiprocessing.pool import Pool +from pathlib import Path +from typing import Callable, NamedTuple + +import numpy as np +import numpy.typing as npt +import pandas as pd +import plotly.graph_objs as go +import scanpy as sc +import sklearn +import sklearn.neighbors +from fast_bioservices import Taxon +from fast_bioservices.pipeline import ensembl_to_gene_id_and_symbol +from loguru import logger +from pandas import DataFrame +from plotly.subplots import make_subplots +from scipy.signal import find_peaks +from sklearn.neighbors import KernelDensity + +from como.custom_types import RNASeqPreparationMethod +from como.migrations import gene_info_migrations +from como.project import Config +from como.utils import convert_gene_data + + +class _FilteringOptions(NamedTuple): + replicate_ratio: float + batch_ratio: float + cut_off: float + high_replicate_ratio: float + high_batch_ratio: float + + +class FilteringTechnique(Enum): + """RNA sequencing filtering capabilities.""" + + cpm = "cpm" + zfpkm = "zfpkm" + tpm = "quantile" + umi = "umi" + + @staticmethod + def from_string(value: str) -> FilteringTechnique: + """Create a filtering technique object from a string.""" + match value.lower(): + case "cpm": + return FilteringTechnique.cpm + case "zfpkm": + return FilteringTechnique.zfpkm + case "quantile": + return FilteringTechnique.tpm + case "umi": + return FilteringTechnique.umi + case _: + possible_values = [t.value for t in FilteringTechnique] + raise ValueError(f"Filtering technique must be one of {possible_values}; got: {value}") + + +class LayoutMethod(Enum): + """RNA sequencing layout method.""" + + paired_end = "paired-end" + single_end = "single-end" + + +@dataclass +class _StudyMetrics: + study: str + num_samples: int + count_matrix: pd.DataFrame + fragment_lengths: npt.NDArray[np.float32] + sample_names: list[str] + layout: list[LayoutMethod] + entrez_gene_ids: list[str] + gene_sizes: npt.NDArray[np.float32] + __normalization_matrix: pd.DataFrame = field(default_factory=pd.DataFrame) + __z_score_matrix: pd.DataFrame = field(default_factory=pd.DataFrame) + __high_confidence_entrez_gene_ids: list[str] = field(default=list) + + def __post_init__(self): + for layout in self.layout: + if layout not in LayoutMethod: + raise ValueError(f"Layout must be 'paired-end' or 'single-end'; got: {layout}") + + @property + def normalization_matrix(self) -> pd.DataFrame: + return self.__normalization_matrix + + @normalization_matrix.setter + def normalization_matrix(self, value: pd.DataFrame) -> None: + self.__normalization_matrix = value + + @property + def z_score_matrix(self) -> pd.DataFrame: + return self.__z_score_matrix + + @z_score_matrix.setter + def z_score_matrix(self, value: pd.DataFrame) -> None: + self.__z_score_matrix = value + + @property + def high_confidence_entrez_gene_ids(self) -> list[str]: + return self.__high_confidence_entrez_gene_ids + + @high_confidence_entrez_gene_ids.setter + def high_confidence_entrez_gene_ids(self, values: list[str]) -> None: + self.__high_confidence_entrez_gene_ids = values + + +Density = namedtuple("Density", ["x", "y"]) + + +class _ZFPKMResult(NamedTuple): + zfpkm: pd.Series + density: Density + mu: float + std_dev: float + max_fpkm: float + + +class _ReadMatrixResults(NamedTuple): + metrics: dict[str, _StudyMetrics] + entrez_gene_ids: list[str] + + +NamedMetrics = dict[str, _StudyMetrics] + + +def k_over_a(k: int, a: float) -> Callable[[npt.NDArray], bool]: + """Return a function that filters rows of an array based on the sum of elements being greater than or equal to A at least k times. + + This code is based on the `kOverA` function found in R's `genefilter` package: https://www.rdocumentation.org/packages/genefilter/versions/1.54.2/topics/kOverA + + :param k: The minimum number of times the sum of elements must be greater than or equal to A. + :param a: The value to compare the sum of elements to. + :return: A function that accepts a NumPy array to perform the actual filtering + """ # noqa: E501 + + def filter_func(row: npt.NDArray) -> bool: + return np.sum(row >= a) >= k + + return filter_func + + +def genefilter(data: pd.DataFrame | npt.NDArray, filter_func: Callable[[npt.NDArray], bool]) -> npt.NDArray: + """Apply a filter function to the rows of the data and return the filtered array. + + This code is based on the `genefilter` function found in R's `genefilter` package: https://www.rdocumentation.org/packages/genefilter/versions/1.54.2/topics/genefilter + + :param data: The data to filter + :param filter_func: THe function to filter the data by + :return: A NumPy array of the filtered data. + """ + match type(data): + case pd.DataFrame: + return data.apply(filter_func, axis=1).values + case npt.NDArray: + return np.apply_along_axis(filter_func, axis=1, arr=data) + case _: + raise ValueError("Unsupported data type. Must be a Pandas DataFrame or a NumPy array.") + + +async def _read_counts_matrix( + *, + context_name: str, + counts_matrix_filepath: Path, + config_filepath: Path, + gene_info_filepath: Path, + taxon_id: Taxon, +) -> _ReadMatrixResults: + """Read the counts matrix and returns the results. + + :param context_name: The context name being processed. Usually a cell type, but can be any string + :param counts_matrix_filepath: The file path to the gene count matrix + :param config_filepath: The file path to the Excel configuration file + :param gene_info_filepath: The file path to gene information generated by `rnaseq_preprocess.py` + :param taxon_id: The NCBI Taxon ID + :return: A dataclass `ReadMatrixResults` + """ + logger.trace(f"Reading config_filepath at '{config_filepath}'") + config_df: pd.DataFrame = pd.read_excel(config_filepath, sheet_name=context_name, header=0) + gene_info: pd.DataFrame = pd.read_csv(gene_info_filepath) + gene_info = gene_info[gene_info["ensembl_gene_id"] != "-"].reset_index(drop=True) + gene_info = gene_info_migrations(gene_info) + + match counts_matrix_filepath.suffix: + case ".csv": + logger.debug(f"Reading CSV file at '{counts_matrix_filepath}'") + counts_matrix: pd.DataFrame = pd.read_csv(counts_matrix_filepath, header=0) + if "ensembl_gene_id" not in counts_matrix.columns: + raise ValueError( + f"Counts matrix must contain a column named 'ensembl_gene_id'. " + f"Ensure the file '{counts_matrix_filepath}' contains this column." + ) + conversion = await ensembl_to_gene_id_and_symbol( + ids=counts_matrix["ensembl_gene_id"].tolist(), taxon=taxon_id + ) + counts_matrix = counts_matrix.merge(conversion, on="ensembl_gene_id", how="left") + + case ".h5ad": + logger.debug(f"Reading h5ad file at '{counts_matrix_filepath}'") + adata: sc.AnnData = sc.read_h5ad(counts_matrix_filepath) + counts_matrix: pd.DataFrame = adata.to_df().T # Make sample names the columns and gene data the index + + # Coherce the incoming gene data (i.e., Gene Symbols) into Entrez and Ensembl Gene IDs + conversion = await convert_gene_data(counts_matrix.index.tolist(), taxon_id) + counts_matrix.index.name = conversion.index.name + counts_matrix = counts_matrix.merge(conversion, left_index=True, right_index=True) + counts_matrix = counts_matrix[counts_matrix["entrez_gene_id"] != "-"] + counts_matrix.reset_index(inplace=True) + + # explicit garbage collection because this function runs for a little while + del adata + del conversion + gc.collect() + + case _: + raise ValueError( + f"Unknown file extension '{counts_matrix_filepath.suffix}'. Valid options are '.csv' or '.h5ad'." + ) + + if counts_matrix.empty: + raise ValueError( + f"Counts matrix is empty. Ensure the file contains data. " + f"Attempted to process file '{counts_matrix_filepath}'" + ) + + # Only include Entrez and Ensembl Gene IDs that are present in `gene_info` + counts_matrix["entrez_gene_id"] = counts_matrix["entrez_gene_id"].str.split("//") + counts_matrix = counts_matrix.explode("entrez_gene_id") + counts_matrix = counts_matrix.replace(to_replace="-", value=pd.NA).dropna() + counts_matrix["entrez_gene_id"] = counts_matrix["entrez_gene_id"].astype(int) + + gene_info = gene_info.replace(to_replace="-", value=pd.NA).dropna() + gene_info["entrez_gene_id"] = gene_info["entrez_gene_id"].astype(int) + + counts_matrix = counts_matrix.merge( + gene_info[["entrez_gene_id", "ensembl_gene_id"]], + on=["entrez_gene_id", "ensembl_gene_id"], + how="inner", + ) + gene_info = gene_info.merge( + counts_matrix[["entrez_gene_id", "ensembl_gene_id"]], + on=["entrez_gene_id", "ensembl_gene_id"], + how="inner", + ) + + entrez_gene_ids: list[str] = gene_info["entrez_gene_id"].tolist() + metrics: NamedMetrics = {} + for study in config_df["study"].unique().tolist(): + study_sample_names = config_df[config_df["study"] == study]["sample_name"].tolist() + layouts = config_df[config_df["study"] == study]["layout"].tolist() + metrics[study] = _StudyMetrics( + count_matrix=counts_matrix[counts_matrix.columns.intersection(study_sample_names)], + fragment_lengths=config_df[config_df["study"] == study]["fragment_length"].values, + sample_names=study_sample_names, + layout=[LayoutMethod(layout) for layout in layouts], + num_samples=len(study_sample_names), + entrez_gene_ids=entrez_gene_ids, + gene_sizes=np.array(gene_info["size"].values).astype(np.float32), + study=study, + ) + metrics[study].fragment_lengths[np.isnan(metrics[study].fragment_lengths)] = 0 + metrics[study].count_matrix.index = pd.Index(entrez_gene_ids, name="entrez_gene_id") + + return _ReadMatrixResults(metrics=metrics, entrez_gene_ids=gene_info["entrez_gene_id"].tolist()) + + +def calculate_tpm(metrics: NamedMetrics) -> NamedMetrics: + """Calculate the Transcripts Per Million (TPM) for each sample in the metrics dictionary.""" + for sample in metrics: + count_matrix = metrics[sample].count_matrix + + gene_sizes = metrics[sample].gene_sizes + + tpm_matrix = pd.DataFrame(data=None, index=count_matrix.index, columns=count_matrix.columns) + for i in range(len(count_matrix.columns)): + values: pd.Series = count_matrix.iloc[:, i] + 1 # Add 1 to prevent division by 0 + rate = np.log(values.tolist()) - np.log(gene_sizes) + denominator = np.log(np.sum(np.exp(rate))) + tpm_value = np.exp(rate - denominator + np.log(1e6)) + tpm_matrix.iloc[:, i] = tpm_value + metrics[sample].normalization_matrix = tpm_matrix + + return metrics + + +def calculate_fpkm(metrics: NamedMetrics) -> NamedMetrics: + """Calculate the Fragments Per Kilobase of transcript per Million mapped reads (FPKM) for each sample in the metrics dictionary.""" # noqa: E501 + matrix_values = [] + for study in metrics: + for sample in range(metrics[study].num_samples): + layout = metrics[study].layout[sample] + count_matrix: npt.NDArray = metrics[study].count_matrix.iloc[:, sample].values + gene_size = metrics[study].gene_sizes + + count_matrix = count_matrix.astype(np.float32) + gene_size = gene_size.astype(np.float32) + + match layout: + case LayoutMethod.paired_end: # FPKM + mean_fragment_lengths = metrics[study].fragment_lengths[sample] + # Ensure non-negative value + effective_length = [max(0, size - (mean_fragment_lengths + 1)) for size in gene_size] + n = count_matrix.sum() + fpkm = ((count_matrix + 1) * 1e9) / (np.array(effective_length) * n) + matrix_values.append(fpkm) + case LayoutMethod.single_end: # RPKM + # Add a pseudocount before log to ensure log(0) does not happen + rate = np.log(count_matrix + 1) - np.log(gene_size) + exp_rate = np.exp(rate - np.log(np.sum(count_matrix)) + np.log(1e9)) + matrix_values.append(exp_rate) + case _: + raise ValueError("Invalid normalization method specified") + + fpkm_matrix = pd.DataFrame(matrix_values).T # Transpose is needed because values were appended as rows + fpkm_matrix = fpkm_matrix[~pd.isna(fpkm_matrix)] + metrics[study].normalization_matrix = fpkm_matrix + + metrics[study].normalization_matrix.columns = metrics[study].count_matrix.columns + + return metrics + + +def _zfpkm_calculation(col: pd.Series, kernel: KernelDensity, peak_parameters: tuple[float, float]) -> _ZFPKMResult: + """Log2 Transformations. + + Stabilize the variance in the data to make the distribution more symmetric; this is helpful for Gaussian fitting + + Kernel Density Estimation (kde) + - Non-parametric method to estimate the probability density function (PDF) of a random variable + - Estimates the distribution of log2-transformed FPKM values + - Bandwidth parameter controls the smoothness of the density estimate + - KDE Explanation + - A way to smooth a histogram to get a better idea of the underlying distribution of the data + - Given a set of data points, we want to understand how they are distributed. + Histograms can be useful, but are sensitive to bin size and number + - The KDE places a "kernel" - a small symmetric function (i.e., Gaussian curve) - at each data point + - The "kernel" acts as a weight, giving more weight to points closer to the center of the kernel, + and less weight to points farther away + - Kernel functions are summed along each point on the x-axis + - A smooth curve is created that represents the estimated density of the data + + Peak Finding + - Identifies that are above a certain height and separated by a minimum distance + - Represent potential local maxima in the distribution + + Peak Selection + - The peak with the highest x-value (from log2-FPKM) is chosen as the mean (mu) + of the "inactive" gene distribution + - The peak representing unexpressed or inactive genes should be at a lower FPKM + value compared to the peak representing expressed genes + + Standard Deviation Estimation + - The mean of log2-FPKM values are greater than the calculated mu + - Standard deviation is estimated based on the assumption that the right tail of the distribution + This represents expressed genes) can be approximated by a half-normal distribution + + zFPKM Transformation + - Centers disbribution around 0 and scales it by the standard deviation. + This makes it easier to compare gene expression across different samples + - Represents the number of standard deviations away from the mean of the "inactive" gene distribution + - Higher zFPKM values indicate higher expression levels relative to the "inactive" peak + - A zFPKM value of 0 represents the mean of the "inactive" distribution + - Research shows that a zFPKM value of -3 or greater can be used as + a threshold for calling a gene as "expressed" + : https://doi.org/10.1186/1471-2164-14-778 + """ + col_log2: npt.NDArray = np.log2(col + 1) + col_log2 = np.nan_to_num(col_log2, nan=0) + refit: KernelDensity = kernel.fit(col_log2.reshape(-1, 1)) # type: ignore + + # kde: KernelDensity = KernelDensity(kernel="gaussian", bandwidth=bandwidth).fit(col_log2.reshape(-1, 1)) + x_range = np.linspace(col_log2.min(), col_log2.max(), 1000) + density = np.exp(refit.score_samples(x_range.reshape(-1, 1))) + peaks, _ = find_peaks(density, height=peak_parameters[0], distance=peak_parameters[1]) + peak_positions = x_range[peaks] + + mu = 0 + max_fpkm = 0 + stddev = 1 + + if len(peaks) != 0: + mu = peak_positions.max() + max_fpkm = density[peaks[np.argmax(peak_positions)]] + u = col_log2[col_log2 > mu].mean() + stddev = (u - mu) * np.sqrt(np.pi / 2) + zfpkm = pd.Series((col_log2 - mu) / stddev, dtype=np.float32, name=col.name) + + return _ZFPKMResult(zfpkm=zfpkm, density=Density(x_range, density), mu=mu, std_dev=stddev, max_fpkm=max_fpkm) + + +def zfpkm_transform( + fpkm_df: pd.DataFrame, + bandwidth: int = 0.5, + peak_parameters: tuple[float, float] = (0.02, 1.0), + update_every_percent: float = 0.1, +) -> tuple[dict[str, _ZFPKMResult], DataFrame]: + """Perform zFPKM calculation/transformation.""" + if update_every_percent > 1: + logger.warning( + f"update_every_percent should be a decimal value between 0 and 1; got: {update_every_percent} - " + f"will convert to percentage" + ) + update_every_percent /= 100 + + total = len(fpkm_df.columns) + update_per_step: int = int(np.ceil(total * update_every_percent)) + cores = multiprocessing.cpu_count() - 2 + logger.debug(f"Processing {total:,} samples through zFPKM transform using {cores} cores") + logger.debug( + f"Will update every {update_per_step:,} steps as this is approximately " + f"{update_every_percent:.1%} of {total:,}" + ) + + with Pool(processes=cores) as pool: + kernel = KernelDensity(kernel="gaussian", bandwidth=bandwidth) + chunksize = int(math.ceil(len(fpkm_df.columns) / (4 * cores))) + partial_func = partial(_zfpkm_calculation, kernel=kernel, peak_parameters=peak_parameters) + chunk_time = time.time() + start_time = time.time() + + log_padding = len(str(f"{total:,}")) + zfpkm_df = pd.DataFrame(data=0, index=fpkm_df.index, columns=fpkm_df.columns) + results: dict[str, _ZFPKMResult] = {} + result: _ZFPKMResult + for i, result in enumerate( + pool.imap( + partial_func, + (fpkm_df[col] for col in fpkm_df.columns), + chunksize=chunksize, + ) + ): + key = str(result.zfpkm.name) + results[key] = result + zfpkm_df[key] = result.zfpkm + + # show updates every X% and at the end, but skip on first iteration + if i != 0 and (i % update_per_step == 0 or i == total): + current_time = time.time() + chunk = current_time - chunk_time + total_time = current_time - start_time + formatted = f"{i:,}" + logger.debug( + f"Processed {formatted:>{log_padding}} of {total:,} - " + f"chunk took {chunk:.1f} seconds - " + f"running for {total_time:.1f} seconds" + ) + chunk_time = current_time + return results, zfpkm_df + + +def zfpkm_plot(results, *, plot_xfloor: int = -4, subplot_titles: bool = True): + """Plot the log2(FPKM) density and fitted Gaussian for each sample. + + :param results: A dictionary of intermediate results from zfpkm_transform. + :param: subplot_titles: Whether to display facet titles (sample names). + :param plot_xfloor: Lower limit for the x-axis. + :param subplot_titles: Whether to display facet titles (sample names). + """ + mega_df = pd.DataFrame(columns=["sample_name", "log2fpkm", "fpkm_density", "fitted_density_scaled"]) + for name, result in results.items(): + stddev = result.std_dev + x = np.array(result.density.x) + y = np.array(result.density.y) + + fitted = np.exp(-0.5 * ((x - result.mu) / stddev) ** 2) / (stddev * np.sqrt(2 * np.pi)) + max_fpkm = y.max() + max_fitted = fitted.max() + scale_fitted = fitted * (max_fpkm / max_fitted) + + df = pd.DataFrame( + { + "sample_name": [name] * len(x), + "log2fpkm": x, + "fpkm_density": y, + "fitted_density_scaled": scale_fitted, + } + ) + mega_df = pd.concat([mega_df, df], ignore_index=True) + + mega_df = mega_df.melt(id_vars=["log2fpkm", "sample_name"], var_name="source", value_name="density") + subplot_titles = list(results.keys()) if subplot_titles else None + fig = make_subplots( + rows=len(results), + cols=1, + subplot_titles=subplot_titles, + vertical_spacing=min(0.05, (1 / (len(results) - 1))), + ) + + for i, (name, group) in enumerate(mega_df.groupby("sample_name"), start=1): + fig.add_trace( + trace=go.Scatter(x=group["log2fpkm"], y=group["density"], mode="lines", name=name, legendgroup=name), + row=i, + col=1, + ) + fig.update_xaxes(title_text="log2(FPKM)", range=[plot_xfloor, max(group["log2fpkm"].tolist())], row=i, col=1) + fig.update_yaxes(title_text="density [scaled]", row=i, col=1) + fig.update_layout(legend_tracegroupgap=0) + + fig.update_layout(height=600 * len(results), width=1000, title_text="zFPKM Plots", showlegend=True) + fig.write_image("zfpkm_plot.png") + + +def calculate_z_score(metrics: NamedMetrics) -> NamedMetrics: + """Calculate the z-score for each sample in the metrics dictionary.""" + for sample in metrics: + log_matrix = np.log(metrics[sample].normalization_matrix) + z_matrix = pd.DataFrame( + data=sklearn.preprocessing.scale(log_matrix, axis=1), columns=metrics[sample].sample_names + ) + metrics[sample].z_score_matrix = z_matrix + return metrics + + +def cpm_filter( + *, + context_name: str, + metrics: NamedMetrics, + filtering_options: _FilteringOptions, + prep: RNASeqPreparationMethod, +) -> NamedMetrics: + """Apply Counts Per Million (CPM) filtering to the count matrix for a given sample.""" + config = Config() + n_exp = filtering_options.replicate_ratio + n_top = filtering_options.high_replicate_ratio + cut_off = filtering_options.cut_off + + sample: str + metric: _StudyMetrics + for sample, metric in metrics.items(): + counts: pd.DataFrame = metric.count_matrix + entrez_ids: list[str] = metric.entrez_gene_ids + library_size: pd.DataFrame = counts.sum(axis=1) + + # For library_sizes equal to 0, add 1 to prevent divide by 0 + # This will not impact the final counts per million calculation because the original counts are still 0 + # thus, (0 / 1) * 1_000_000 = 0 + library_size[library_size == 0] = 1 + + output_filepath = config.result_dir / context_name / prep.value / f"CPM_Matrix_{prep.value}_{sample}.csv" + output_filepath.parent.mkdir(parents=True, exist_ok=True) + counts_per_million: pd.DataFrame = (counts / library_size) * 1_000_000 + counts_per_million.insert(0, "entrez_gene_ids", pd.Series(entrez_ids)) + logger.debug(f"Writing CPM matrix to {output_filepath}") + counts_per_million.to_csv(output_filepath, index=False) + + # TODO: Counts per million is adding ~61,500 columns (equal to the number of genes) for some reason. + # Most likely due to multiplying by 1_000_000, not exactly sure why + + min_samples = round(n_exp * len(counts.columns)) # noqa: F841 + top_samples = round(n_top * len(counts.columns)) # noqa: F841 + test_bools = pd.DataFrame({"entrez_gene_ids": entrez_ids}) + for i in range(len(counts_per_million.columns)): + cutoff = ( + 10e6 / (np.median(np.sum(counts[:, i]))) + if cut_off == "default" + else 1e6 * cut_off / np.median(np.sum(counts[:, i])) + ) + test_bools = test_bools.merge(counts_per_million[counts_per_million.iloc[:, i] > cutoff]) + + return metrics + + +def tpm_quantile_filter(*, metrics: NamedMetrics, filtering_options: _FilteringOptions) -> NamedMetrics: + """Apply quantile-based filtering to the TPM matrix for a given sample.""" + # TODO: Write the TPM matrix to disk + + n_exp = filtering_options.replicate_ratio + n_top = filtering_options.high_replicate_ratio + cut_off = filtering_options.cut_off + metrics = calculate_tpm(metrics) + + sample: str + metric: _StudyMetrics + for sample, metric in metrics.items(): + entrez_ids = metric.entrez_gene_ids + gene_size = metric.gene_sizes + tpm_matrix: pd.DataFrame = metric.normalization_matrix + + min_samples = round(n_exp * len(tpm_matrix.columns)) + top_samples = round(n_top * len(tpm_matrix.columns)) + + tpm_quantile = tpm_matrix[tpm_matrix > 0] + quantile_cutoff = np.quantile( + a=tpm_quantile.values, q=1 - (cut_off / 100), axis=0 + ) # Compute quantile across columns + boolean_expression = pd.DataFrame( + data=tpm_matrix > quantile_cutoff, index=tpm_matrix.index, columns=tpm_matrix.columns + ).astype(int) + + min_func = k_over_a(min_samples, 0.9) + top_func = k_over_a(top_samples, 0.9) + + min_genes: npt.NDArray[bool] = genefilter(boolean_expression, min_func) + top_genes: npt.NDArray[bool] = genefilter(boolean_expression, top_func) + + # Only keep `entrez_gene_ids` that pass `min_genes` + metric.entrez_gene_ids = [gene for gene, keep in zip(entrez_ids, min_genes) if keep] + metric.gene_sizes = [gene for gene, keep in zip(gene_size, min_genes) if keep] + metric.count_matrix = metric.count_matrix.iloc[min_genes, :] + metric.normalization_matrix = metrics[sample].normalization_matrix.iloc[min_genes, :] + + keep_top_genes = [gene for gene, keep in zip(entrez_ids, top_genes) if keep] + metric.high_confidence_entrez_gene_ids = [gene for gene, keep in zip(entrez_ids, keep_top_genes) if keep] + + metrics = calculate_z_score(metrics) + + return metrics + + +def zfpkm_filter(*, metrics: NamedMetrics, filtering_options: _FilteringOptions, calcualte_fpkm: bool) -> NamedMetrics: + """Apply zFPKM filtering to the FPKM matrix for a given sample.""" + min_sample_expression = filtering_options.replicate_ratio + high_confidence_sample_expression = filtering_options.high_replicate_ratio + cut_off = filtering_options.cut_off + + if calcualte_fpkm: + metrics = calculate_fpkm(metrics) + + metric: _StudyMetrics + for metric in metrics.values(): + # if fpkm was not calculated, the normalization matrix will be empty; collect the count matrix instead + matrix = metric.count_matrix if metric.normalization_matrix.empty else metric.normalization_matrix + matrix = matrix[matrix.sum(axis=1) > 0] + + minimums = matrix == 0 + results, zfpkm_df = zfpkm_transform(matrix) + zfpkm_df[minimums] = -4 + zfpkm_plot(results) + + # determine which genes are expressed + min_samples = round(min_sample_expression * len(zfpkm_df.columns)) + min_func = k_over_a(min_samples, cut_off) + min_genes: npt.NDArray[bool] = genefilter(zfpkm_df, min_func) + metric.entrez_gene_ids = [gene for gene, keep in zip(metric.entrez_gene_ids, min_genes) if keep] + + # determine which genes are confidently expressed + top_samples = round(high_confidence_sample_expression * len(zfpkm_df.columns)) + top_func = k_over_a(top_samples, cut_off) + top_genes: npt.NDArray[bool] = genefilter(zfpkm_df, top_func) + metric.high_confidence_entrez_gene_ids = [gene for gene, keep in zip(metric.entrez_gene_ids, top_genes) if keep] + + return metrics + + +def filter_counts( + *, + context_name: str, + metrics: NamedMetrics, + technique: FilteringTechnique, + filtering_options: _FilteringOptions, + prep: RNASeqPreparationMethod, +) -> NamedMetrics: + """Filter the count matrix based on the specified technique.""" + match technique: + case FilteringTechnique.cpm: + return cpm_filter( + context_name=context_name, metrics=metrics, filtering_options=filtering_options, prep=prep + ) + case FilteringTechnique.tpm: + return tpm_quantile_filter(metrics=metrics, filtering_options=filtering_options) + case FilteringTechnique.zfpkm: + return zfpkm_filter(metrics=metrics, filtering_options=filtering_options, calcualte_fpkm=True) + case FilteringTechnique.umi: + return zfpkm_filter(metrics=metrics, filtering_options=filtering_options, calcualte_fpkm=False) + case _: + raise ValueError(f"Technique must be one of {FilteringTechnique}") + + +async def save_rnaseq_tests( + context_name: str, + counts_matrix_filepath: Path, + config_filepath: Path, + gene_info_filepath: Path, + output_filepath: Path, + prep: RNASeqPreparationMethod, + taxon_id: Taxon, + replicate_ratio: float, + batch_ratio: float, + high_replicate_ratio: float, + high_batch_ratio: float, + technique: FilteringTechnique, + cut_off: int | float, +): + """Save the results of the RNA-Seq tests to a CSV file.""" + filtering_options = _FilteringOptions( + replicate_ratio=replicate_ratio, + batch_ratio=batch_ratio, + cut_off=cut_off, + high_replicate_ratio=high_replicate_ratio, + high_batch_ratio=high_batch_ratio, + ) + + if prep == RNASeqPreparationMethod.SCRNA: + technique = FilteringTechnique.umi + logger.warning( + "Single cell filtration does not normalize and assumes " + "gene counts are counted with Unique Molecular Identifiers (UMIs). " + "Setting filtering technique to UMI now." + ) + + read_counts_results: _ReadMatrixResults = await _read_counts_matrix( + context_name=context_name, + counts_matrix_filepath=counts_matrix_filepath, + config_filepath=config_filepath, + gene_info_filepath=gene_info_filepath, + taxon_id=taxon_id, + ) + metrics = read_counts_results.metrics + entrez_gene_ids = read_counts_results.entrez_gene_ids + + metrics = filter_counts( + context_name=context_name, + metrics=metrics, + technique=technique, + filtering_options=filtering_options, + prep=prep, + ) + + expressed_genes: list[str] = [] + top_genes: list[str] = [] + for metric in metrics.values(): + expressed_genes.extend(metric.entrez_gene_ids) + top_genes.extend(metric.high_confidence_entrez_gene_ids) + + expression_frequency = pd.Series(expressed_genes).value_counts() + expression_df = pd.DataFrame( + {"entrez_gene_id": expression_frequency.index, "frequency": expression_frequency.values} + ) + expression_df["prop"] = expression_df["frequency"] / len(metrics) + expression_df = expression_df[expression_df["prop"] >= filtering_options.batch_ratio] + + top_frequency = pd.Series(top_genes).value_counts() + top_df = pd.DataFrame({"entrez_gene_id": top_frequency.index, "frequency": top_frequency.values}) + top_df["prop"] = top_df["frequency"] / len(metrics) + top_df = top_df[top_df["prop"] >= filtering_options.high_batch_ratio] + + boolean_matrix = pd.DataFrame(data={"entrez_gene_id": entrez_gene_ids, "expressed": 0, "high": 0}) + for gene in entrez_gene_ids: + if gene in expression_df["entrez_gene_id"]: + boolean_matrix.loc[gene, "expressed"] = 1 + if gene in top_df["entrez_gene_id"]: + boolean_matrix.loc[gene, "high"] = 1 + + expressed_count = len(boolean_matrix[boolean_matrix["expressed"] == 1]) + high_confidence_count = len(boolean_matrix[boolean_matrix["high"] == 1]) + + boolean_matrix.to_csv(output_filepath, index=False) + logger.info( + f"{context_name} - Found {expressed_count} expressed and {high_confidence_count} confidently expressed genes" + ) + logger.success(f"Wrote boolean matrix to {output_filepath}") diff --git a/main/como/rnaseq_gen.py b/main/como/rnaseq_gen.py index db3150ee..8f8ffa34 100644 --- a/main/como/rnaseq_gen.py +++ b/main/como/rnaseq_gen.py @@ -1,196 +1,189 @@ -#!/usr/bin/python3 +from __future__ import annotations import argparse -import re -from enum import Enum -from pathlib import Path -from typing import Optional +import asyncio +from dataclasses import dataclass import pandas as pd -from rpy2.robjects import pandas2ri - -from como import rpy2_api -from como.project import Config - -# enable r to py conversion -pandas2ri.activate() - -r_file_path = Path(__file__).parent / "rscripts" / "rnaseq.R" - - -def load_rnaseq_tests(filename, context_name, lib_type): - """ - Load rnaseq results returning a dictionary of test (context, context, cell, etc ) names and rnaseq expression data - """ - config = Config() - - def load_dummy_dict(): - dat = pd.read_csv(config.data_dir / "data_matrices" / "placeholder" / "placeholder_empty_data.csv", index_col="ENTREZ_GENE_ID") - return "dummy", dat - - if not filename or filename == "None": # not using this data type, use empty dummy data matrix - return load_dummy_dict() - - inquiry_full_path = config.data_dir / "config_sheets" / filename - if not inquiry_full_path.exists(): # check that config file exist (isn't needed to load, but helps user) - raise FileNotFoundError(f"Error: Config file not found at {inquiry_full_path}") - - if lib_type == "total": # if using total RNA-seq library prep - filename = f"rnaseq_total_{context_name}.csv" - elif lib_type == "mrna": # if using mRNA-seq library prep - filename = f"rnaseq_mrna_{context_name}.csv" - elif lib_type == "scrna": # if using single-cell RNA-seq - filename = f"rnaseq_scrna_{context_name}.csv" - else: - raise ValueError(f"Unsupported RNA-seq library type: {lib_type}. Must be one of 'total', 'mrna', 'sc'.") - - save_filepath = config.result_dir / context_name / lib_type / filename - if save_filepath.exists(): - data = pd.read_csv(save_filepath, index_col="ENTREZ_GENE_ID") - print(f"Read from {save_filepath}") - return context_name, data - - else: - print( - f"{lib_type} gene expression file for {context_name} was not found at {save_filepath}. This may be " - f"intentional. Contexts where {lib_type} data can be found in /work/data/results/{context_name}/ will " - "still be used if found for other contexts." - ) - return load_dummy_dict() - - -def handle_context_batch( - config_filename, - replicate_ratio, - batch_ratio, - replicate_ratio_high, - batch_ratio_high, - technique, - quantile, - min_count, - min_zfpkm, - prep, -): - """ - Handle iteration through each context type and create rnaseq expression file by calling rnaseq.R +from fast_bioservices import Taxon +from loguru import logger + +from como import Config +from como.custom_types import RNASeqPreparationMethod +from como.rnaseq import FilteringTechnique, save_rnaseq_tests + + +@dataclass +class _Arguments: + config_file: str + replicate_ratio: float + batch_ratio: float + high_replicate_ratio: float + high_batch_ratio: float + filtering_technique: FilteringTechnique + minimum_cutoff: int | str + library_prep: RNASeqPreparationMethod + taxon: Taxon + + def __post_init__(self): + self.library_prep = RNASeqPreparationMethod.from_string(str(self.library_prep)) + self.filtering_technique = FilteringTechnique.from_string(str(self.filtering_technique)) + + if self.minimum_cutoff is None: + if self.filtering_technique == FilteringTechnique.tpm: + self.minimum_cutoff = 25 + elif self.filtering_technique == FilteringTechnique.cpm: + self.minimum_cutoff = "default" + elif self.filtering_technique == FilteringTechnique.zfpkm: + self.minimum_cutoff = -3 + + +async def _handle_context_batch( + config_filename: str, + replicate_ratio: float, + batch_ratio: float, + replicate_ratio_high: float, + batch_ratio_high: float, + technique: FilteringTechnique, + cut_off: int | float | str, + prep: RNASeqPreparationMethod, + taxon: Taxon, +) -> None: + """Iterate through each context type and create rnaseq expression file. + + :param config_filename: The configuration filename to read + :param replicate_ratio: The percentage of replicates that a gene must + appear in for a gene to be marked as "active" in a batch/study + :param batch_ratio: The percentage of batches that a gene must appear in for a gene to be marked as 'active" + :param replicate_ratio_high: The percentage of replicates that a gene must + appear in for a gene to be marked "highly confident" in its expression in a batch/study + :param batch_ratio_high: The percentage of batches that a gene must + appear in for a gene to be marked "highly confident" in its expression + :param technique: The filtering technique to use + :param cut_off: The cutoff value to use for the provided filtering technique + :param prep: The library preparation method + :param taxon: The NCBI Taxon ID + :return: None """ config = Config() config_filepath = config.config_dir / config_filename + if not config_filepath.exists(): + raise FileNotFoundError(f"Unable to find '{config_filename}' at the path: '{config_filepath}'") xl = pd.ExcelFile(config_filepath) sheet_names = xl.sheet_names - print(f"Reading config file: {config_filepath}") + logger.info(f"Reading config file: {config_filepath}") for context_name in sheet_names: - print(f"\nStarting '{context_name}'") + logger.debug(f"Starting '{context_name}'") + + rnaseq_input_filepath = ( + config.data_dir / "data_matrices" / context_name / f"gene_counts_matrix_{prep.value}_{context_name}" + ) + if prep == RNASeqPreparationMethod.SCRNA: + rnaseq_input_filepath = rnaseq_input_filepath.with_suffix(".h5ad") + elif prep in {RNASeqPreparationMethod.TOTAL, RNASeqPreparationMethod.MRNA}: + rnaseq_input_filepath = rnaseq_input_filepath.with_suffix(".csv") - rnaseq_input_filepath = config.data_dir / "data_matrices" / context_name / f"gene_counts_matrix_{prep}_{context_name}.csv" if not rnaseq_input_filepath.exists(): - print(f"Gene counts matrix not found at {rnaseq_input_filepath}, skipping...") + logger.warning(f"Gene counts matrix not found at {rnaseq_input_filepath}, skipping...") continue gene_info_filepath = config.data_dir / "gene_info.csv" - rnaseq_output_filepath = config.result_dir / context_name / prep / f"rnaseq_{prep}_{context_name}.csv" + rnaseq_output_filepath = ( + config.result_dir / context_name / prep.value / f"rnaseq_{prep.value}_{context_name}.csv" + ) rnaseq_output_filepath.parent.mkdir(parents=True, exist_ok=True) - print(f"Gene info:\t\t{gene_info_filepath}") - print(f"Count matrix:\t\t{rnaseq_input_filepath}") - - rpy2_api.Rpy2( - r_file_path=r_file_path, - counts_matrix_file=rnaseq_input_filepath.as_posix(), - config_file=config_filepath.as_posix(), - out_file=rnaseq_output_filepath.as_posix(), - info_file=gene_info_filepath.as_posix(), + await save_rnaseq_tests( context_name=context_name, + counts_matrix_filepath=rnaseq_input_filepath, + config_filepath=config_filepath, + output_filepath=rnaseq_output_filepath.as_posix(), + gene_info_filepath=gene_info_filepath, prep=prep, replicate_ratio=replicate_ratio, batch_ratio=batch_ratio, - replicate_ratio_high=replicate_ratio_high, - batch_ratio_high=batch_ratio_high, + high_replicate_ratio=replicate_ratio_high, + high_batch_ratio=batch_ratio_high, technique=technique, - quantile=quantile, - min_count=min_count, - min_zfpkm=min_zfpkm, - ).call_function("save_rnaseq_tests") - - print(f"Results saved at:\t{rnaseq_output_filepath}") - - -class Technique(Enum): - ZFPKM = "zfpkm" - TPM = "quantile" - CPM = "cpm" + cut_off=cut_off, + taxon_id=taxon, + ) + logger.success(f"Results saved at '{rnaseq_output_filepath}'") -def rnaseq_gen( +async def rnaseq_gen( + # config_filepath: Path, config_filename: str, + prep: RNASeqPreparationMethod, + taxon_id: int | str | Taxon, replicate_ratio: float = 0.5, + high_replicate_ratio: float = 1.0, batch_ratio: float = 0.5, - replicate_ratio_high: float = 1.0, - batch_ratio_high: float = 1.0, - technique: Technique = Technique.TPM, - cut_off: Optional[int] = None, - prep: Optional[str] = "", + high_batch_ratio: float = 1.0, + technique: FilteringTechnique | str = FilteringTechnique.tpm, + cut_off: int | float | None = None, ) -> None: - if technique not in Technique: - raise ValueError(f"Technique must be one of {Technique}") - - if technique == Technique.TPM: - if cut_off is None: - cut_off = 25 - - if cut_off < 1 or cut_off > 100: - raise ValueError("Quantile must be between 1 - 100") - - elif technique == Technique.CPM: - if cut_off is not None and cut_off < 0: - raise ValueError("Cutoff must be greater than 0") - - if cut_off is None: - cut_off = "default" - elif technique == Technique.ZFPKM: - # if cut_off is not None and (cut_off < -3 or cut_off > -2): - # raise ValueError("Cutoff must be between -3 and -2") - - if cut_off is None: - cut_off = "default" - - prep = prep.replace(" ", "") - - handle_context_batch( - config_filename, - replicate_ratio, - batch_ratio, - replicate_ratio_high, - batch_ratio_high, - technique.value, - cut_off, - cut_off, - cut_off, - prep, + """Generate a list of active and high-confidence genes from a gene count matrix. + + Replicates are compared for consensus within the study/batch number according to replicate ratios, + then study/batch numbers are checked for consensus according to batch ratios. + The zFPKM method is outlined here: https://pubmed.ncbi.nlm.nih.gov/24215113/ + + :param config_filename: The configuration filename to read + :param prep: The preparation method + :param taxon_id: The NCBI Taxon ID + :param replicate_ratio: The percentage of replicates that a gene must + appear in for a gene to be marked as "active" in a batch/study + :param batch_ratio: The percentage of batches that a gene must appear in for a gene to be marked as 'active" + :param high_replicate_ratio: The percentage of replicates that a gene must + appear in for a gene to be marked "highly confident" in its expression in a batch/study + :param high_batch_ratio: The percentage of batches that a gene must + appear in for a gene to be marked "highly confident" in its expression + :param technique: The filtering technique to use + :param cut_off: The cutoff value to use for the provided filtering technique + :return: None + """ + if isinstance(technique, str): + technique = FilteringTechnique(technique.lower()) + if isinstance(taxon_id, (str, int)): + taxon_id = Taxon.from_string(str(taxon_id)) + + match technique: + case FilteringTechnique.tpm: + cut_off = 25 if cut_off is None else cut_off + if cut_off < 1 or cut_off > 100: + raise ValueError("Quantile must be between 1 - 100") + + case FilteringTechnique.cpm: + if cut_off is not None and cut_off < 0: + raise ValueError("Cutoff must be greater than 0") + elif cut_off is None: + cut_off = "default" + + case FilteringTechnique.zfpkm: + cut_off = "default" if cut_off is None else cut_off + case FilteringTechnique.umi: + pass + case _: + raise ValueError(f"Technique must be one of {FilteringTechnique}") + + await _handle_context_batch( + config_filename=config_filename, + replicate_ratio=replicate_ratio, + replicate_ratio_high=high_replicate_ratio, + batch_ratio=batch_ratio, + batch_ratio_high=high_batch_ratio, + technique=technique, + cut_off=cut_off, + prep=prep, + taxon=taxon_id, ) -def main(): - """ - Generate a list of active and high-confidence genes from a counts matrix using a user defined - at normalization-technique at /work/data/results//rnaseq_.csv - Currently, can filter raw RNA-seq counts using three normalization techniques. Which are defined in rnaseq.R - TPM Quantile, where each replicate is normalized with Transcripts-per-million and an upper quantile is taken to - create a boolean list of active genes for the replicate. Replicates are compared for consensus within the - study/batch number according to user-defined ratios and then study/batch numbers are checked for consensus - according to different user defined ratios. **CITATION NEEDED** **Recomended if user wants more control over the - size of the model, like a smaller model that allows for only the most expressed reactions, or a larger more - encompassing one that contains less essential reactions. - zFPKM method outlined in: https://pubmed.ncbi.nlm.nih.gov/24215113/ can be used. Counts will be normalized using - zFPKM and genes > -3 will be considered expressed per thier recommendation. Expressed genes will be checked for - consensus at the replicate and study/batch levels the same as TPM Quantile. **Recommended if user wants to give - least input over gene essentially determination and use the most standardized method of active gene determination. - flat cutoff of CPM (counts per million) normalized values, check for consensus the same as other methods. - """ - +def _parse_args() -> _Arguments: parser = argparse.ArgumentParser( prog="rnaseq_gen.py", description="Generate a list of active and high-confidence genes from a counts matrix using a user defined " @@ -204,7 +197,7 @@ def main(): "--config-file", type=str, required=True, - dest="config_filename", + dest="config_file", help="Name of config .xlsx file in the /work/data/config_files/. Can be generated using " "rnaseq_preprocess.py or manually created and imported into the Juypterlab", ) @@ -236,10 +229,11 @@ def main(): type=float, required=False, default=1.0, - dest="replicate_ratio_high", + dest="high_replicate_ratio", help="Ratio of replicates required for a gene to be considered high-confidence. " "High-confidence genes ignore consensus with other data-sources, such as proteomics. " - "Example: 0.9 means that for a gene to be high-confidence, at least 90% of replicates in a group must pass the cutoff after normalization", + "Example: 0.9 means that for a gene to be high-confidence, " + "at least 90% of replicates in a group must pass the cutoff after normalization", ) parser.add_argument( "-gh", @@ -247,100 +241,70 @@ def main(): type=float, required=False, default=1.0, - dest="batch_ratio_high", - help="Ratio of groups (studies/batches) required for a gene to be considered high-confidence within that group. " + dest="high_batch_ratio", + help="Ratio of studies/batches required for a gene to be considered high-confidence within that group. " "High-confidence genes ignore consensus with other data-sources, like proteomics. " - "Example: 0.9 means that for a gene to be high-confidence, at least 90% of groups in a study must have passed the replicate ratio test", + "Example: 0.9 means that for a gene to be high-confidence, " + "at least 90% of groups in a study must have passed the replicate ratio test", + ) + parser.add_argument( + "--taxon", + "--taxon-id", + type=str, + required=True, + dest="taxon", + help="The NCBI Taxonomy ID that is being proessed. '9606' for humans, '10090' for mice.", ) parser.add_argument( "-t", "--filt-technique", type=str, required=False, - default="quantile-tpm", - dest="technique", - help="Technique to normalize and filter counts with. Either 'zfpkm', 'quantile-tpm' or " - "'flat-cpm'. More info about each method is discussed in pipeline.ipynb.", + default="quantile", + dest="filtering_technique", + help="Technique to normalize and filter counts with. " + "Either 'zfpkm', 'quantile', or 'cpm'. More info about each method is discussed in pipeline.ipynb.", ) parser.add_argument( - "-q", - "--quantile", + "--minimum-cutoff", type=int, required=False, - default=25, - dest="quantile", - help="Cutoff used for quantile-tpm normalization and filtration technique. Example: 25 means " - "that genes with TPM > 75% percentile wik=ll be considered active for that replicate.", - ) - parser.add_argument( - "-m", - "--min-count", - required=False, - default="default", - dest="min_count", - help="Cutoff used for cpm. Minimum number of counts to be considered expressed, alternatively " - "use 'default' to use method outlined in CITATION NEEDED", - ) - parser.add_argument( - "-z", - "--min-zfpkm", - required=False, - default="default", - dest="min_zfpkm", - help="Cutoff used for zfpkm. Minimum zfpkm to be considered expressed, according to the paper, " - "CITATION NEEDED, should be between -3 and -2.", + default=None, + dest="minimum_cutoff", + help="The minimum cutoff used for the filtration technique. " + "If the filtering technique is zFPKM, the default is -3. " + "If the filtering technique is quantile-tpm, the default is 25. " + "If the filtering technique is flat-cpm, the default is determined dynamically. " + "If the filtering technique is quantile, the default is 25.", ) parser.add_argument( "-p", "--library-prep", - required=False, - default="", - dest="prep", - help="Library preparation used, will separate samples into groups to only compare similarly " - "prepared libraries. For example, mRNA, total-rna, scRNA, etc", + required=True, + choices=["total", "mrna", "scrna"], + dest="library_prep", + help="Library preparation method. " + "Will separate samples into groups to only compare similarly prepared libraries. " + "For example, mRNA, total-rna, scRNA, etc", ) - args = parser.parse_args() - - config_filename = args.config_filename - replicate_ratio = args.replicate_ratio - batch_ratio = args.batch_ratio - replicate_ratio_high = args.replicate_ratio_high - batch_ratio_high = args.batch_ratio_high - technique = args.technique - quantile = args.quantile - min_count = args.min_count - prep = args.prep - min_zfpkm = args.min_zfpkm - - if re.search("tpm", technique.lower()) or re.search("quantile", technique.lower()): - technique = "quantile" - elif re.search("cpm", technique.lower()): - technique = "cpm" - elif re.search("zfpkm", technique.lower()): - technique = "zfpkm" - else: - raise ValueError("Technique not recognized. Must be 'tpm-quantile', 'cpm', or 'zfpkm'.") - - if int(quantile) > 100 or int(quantile) < 1: - print("Quantile must be between 1 - 100") - - prep = prep.replace(" ", "") - - handle_context_batch( - config_filename, - replicate_ratio, - batch_ratio, - replicate_ratio_high, - batch_ratio_high, - technique, - quantile, - min_count, - min_zfpkm, - prep, - ) - print("\nDone!") + args.filtering_technique = args.filtering_technique.lower() + args.taxon = Taxon.from_int(int(args.taxon)) if str(args.taxon).isdigit() else Taxon.from_string(str(args.taxon)) # type: ignore + return _Arguments(**vars(args)) if __name__ == "__main__": - main() + args = _parse_args() + asyncio.run( + rnaseq_gen( + config_filename=args.config_file, + replicate_ratio=args.replicate_ratio, + batch_ratio=args.batch_ratio, + high_replicate_ratio=args.high_replicate_ratio, + high_batch_ratio=args.high_batch_ratio, + technique=args.filtering_technique, + cut_off=args.minimum_cutoff, + prep=args.library_prep, + taxon_id=args.taxon, + ) + ) diff --git a/main/como/rnaseq_preprocess.py b/main/como/rnaseq_preprocess.py index 51cdd759..c522f0f1 100644 --- a/main/como/rnaseq_preprocess.py +++ b/main/como/rnaseq_preprocess.py @@ -1,62 +1,315 @@ +from __future__ import annotations + import argparse +import asyncio import re -import sys +from dataclasses import dataclass, field from pathlib import Path -from typing import Optional, Union +from typing import Literal import numpy as np import pandas as pd -from fast_bioservices import BioDBNet, Input, Output, Taxon +import scanpy as sc +from fast_bioservices import Input, Taxon +from fast_bioservices.biothings.mygene import MyGene +from loguru import logger + +from como import Config, stringlist_to_list +from como.utils import convert_gene_data + + +@dataclass +class _Arguments: + context_names: list[str] + taxon_id: Taxon | int | str + mode: Literal["create", "provide"] + input_format: str + provided_matrix_fname: str = None + + +@dataclass +class _STARinformation: + num_unmapped: list[int] + num_multimapping: list[int] + num_no_feature: list[int] + num_ambiguous: list[int] + gene_names: list[str] + count_matrix: pd.DataFrame + + @property + def num_genes(self): + return len(self.count_matrix) + + @classmethod + def build_from_tab(cls, filepath: Path) -> _STARinformation: + if filepath.suffix != ".tab": + raise ValueError(f"Building STAR information requires a '.tab' file; received: '{filepath}'") + with filepath.open("r") as i_stream: + num_unmapped = [int(i) for i in next(i_stream).rstrip("\n").split("\t")[1:]] + num_multimapping = [int(i) for i in next(i_stream).rstrip("\n").split("\t")[1:]] + num_no_feature = [int(i) for i in next(i_stream).rstrip("\n").split("\t")[1:]] + num_ambiguous = [int(i) for i in next(i_stream).rstrip("\n").split("\t")[1:]] + + df = pd.read_csv( + filepath, + sep="\t", + skiprows=4, + names=[ + "ensembl_gene_id", + "unstranded_rna_counts", + "first_read_transcription_strand", + "second_read_transcription_strand", + ], + ) + # Remove NA values + df = df[~df["ensembl_gene_id"].isna()] + return _STARinformation( + num_unmapped=num_unmapped, + num_multimapping=num_multimapping, + num_no_feature=num_no_feature, + num_ambiguous=num_ambiguous, + gene_names=df["ensembl_gene_id"].values.tolist(), + count_matrix=df, + ) -from como import como_utilities, rpy2_api -from como.project import Config -r_file_path: Path = Path(__file__).parent / "rscripts" / "generate_counts_matrix.R" +@dataclass +class _StudyMetrics: + study_name: str + count_files: list[Path] + strandedness_files: list[Path] + __sample_names: list[str] = field(default_factory=list) + __num_samples: int = 0 + @property + def sample_names(self) -> list[str]: + return self.__sample_names -def create_counts_matrix(context_name): - """ - Create a counts matrix by reading gene counts tables in COMO_input///geneCounts/ - Uses R in backend (generate_counts_matrix.R) - """ - config = Config() - input_dir = config.data_dir / "COMO_input" / context_name - print(f"Looking for STAR gene count tables in '{input_dir}'") + @property + def num_samples(self): + return self.__num_samples + + def __post_init__(self): + self.__num_samples = len(self.count_files) + self.__sample_names = [f.stem for f in self.count_files] + + if len(self.count_files) != len(self.strandedness_files): + raise ValueError( + f"Unequal number of count files and strand files for study '{self.study_name}'. " + f"Found {len(self.count_files)} count files and {len(self.strandedness_files)} strand files." + ) + + if self.num_samples != len(self.count_files): + raise ValueError( + f"Unequal number of samples and count files for study '{self.study_name}'. " + f"Found {self.num_samples} samples and {len(self.count_files)} count files." + ) + + if self.num_samples != len(self.strandedness_files): + raise ValueError( + f"Unequal number of samples and strand files for study '{self.study_name}'. " + f"Found {self.num_samples} samples and {len(self.strandedness_files)} strand files." + ) + + if self.__num_samples == 1: + raise ValueError(f"Only one sample exists for study {self.study_name}. Provide at least two samples") + + self.count_files.sort() + self.strandedness_files.sort() + self.__sample_names.sort() + + +def _context_from_filepath(file: Path) -> str: + return file.stem.split("_S")[0] + + +def _sample_name_from_filepath(file: Path) -> str: + return re.search(r".+_S\d+R\d+", file.stem).group() + + +def _organize_gene_counts_files(data_dir: Path) -> list[_StudyMetrics]: + root_gene_count_dir = Path(data_dir, "geneCounts") + root_strandedness_dir = Path(data_dir, "strandedness") + + gene_counts_directories: list[Path] = sorted( + [p for p in Path(root_gene_count_dir).glob("*") if not p.name.startswith(".")] + ) + strandedness_directories: list[Path] = sorted( + [p for p in Path(root_strandedness_dir).glob("*") if not p.name.startswith(".")] + ) + + if len(gene_counts_directories) != len(strandedness_directories): + raise ValueError( + f"Unequal number of gene count directories and strandedness directories. " + f"Found {len(gene_counts_directories)} gene count directories and {len(strandedness_directories)} strandedness directories." # noqa: E501 + f"\nGene count directory: {root_gene_count_dir}\nStrandedness directory: {root_strandedness_dir}" + ) + + # For each study, collect gene count files, fragment files, insert size files, layouts, and strandedness information + study_metrics: list[_StudyMetrics] = [] + for gene_dir, strand_dir in zip(gene_counts_directories, strandedness_directories): + if gene_dir.stem != strand_dir.stem: + raise ValueError( + f"Gene directory name of '{gene_dir.stem}' does not match stranded directory name of '{strand_dir.stem}'" # noqa: E501 + ) + + study_metrics.append( + _StudyMetrics( + study_name=gene_dir.stem, + count_files=list(gene_dir.glob("*.tab")), + strandedness_files=list(strand_dir.glob("*.txt")), + ) + ) + return study_metrics + + +def _process_first_multirun_sample(strand_file: Path, all_counts_files: list[Path]): + sample_count = pd.DataFrame() + for file in all_counts_files: + star_information = _STARinformation.build_from_tab(file) + strand_information = strand_file.read_text().rstrip("\n").lower() + + if strand_information not in ("none", "first_read_transcription_strand", "second_read_transcription_strand"): + raise ValueError( + f"Unrecognized Strand Information: {strand_information}; " + f"expected 'none', 'first_read_transcription_strand', or 'second_read_transcription_strand'" + ) + + if strand_information == "none": + strand_information = "unstranded_rna_counts" + + run_counts = star_information.count_matrix[["gene_id", strand_information]] + run_counts.columns = pd.Index(["ensembl_gene_id", "counts"]) + if sample_count.empty: + sample_count = run_counts + else: + # Merge to take all items from both data frames + sample_count = sample_count.merge(run_counts, on="ensembl_gene_id", how="outer") + + # Set na values to 0 + sample_count = sample_count.fillna(value="0") + sample_count.iloc[:, 1:] = sample_count.iloc[:, 1:].apply(pd.to_numeric) + + count_sums: pd.DataFrame = pd.DataFrame(sample_count.sum(axis=1, numeric_only=True)) + count_sums.insert(0, "ensembl_gene_id", sample_count["ensembl_gene_id"]) + count_sums.columns = pd.Index(["ensembl_gene_id", _sample_name_from_filepath(strand_file)]) + return count_sums + + +def _process_standard_replicate(counts_file: Path, strand_file: Path, sample_name: str): + star_information = _STARinformation.build_from_tab(counts_file) + strand_information = strand_file.read_text().rstrip("\n").lower() + + if strand_information not in ("none", "first_read_transcription_strand", "second_read_transcription_strand"): + raise ValueError( + f"Unrecognized Strand Information: {strand_information}; " + f"expected 'none', 'first_read_transcription_strand', or 'second_read_transcription_strand'" + ) + + if strand_information == "none": + strand_information = "unstranded_rna_counts" + + sample_count = star_information.count_matrix[["ensembl_gene_id", strand_information]] + sample_count.columns = pd.Index(["ensembl_gene_id", sample_name]) + return sample_count + + +def _prepare_sample_counts( + sample_name: str, + counts_file: Path, + strand_file: Path, + all_counts_files: list[Path], +) -> pd.DataFrame | Literal["SKIP"]: + # Test if the counts_file is the first run in a multi-run smaple + if re.search(r"R\d+r1", counts_file.as_posix()): + return _process_first_multirun_sample(strand_file=strand_file, all_counts_files=all_counts_files) + elif re.search(r"R\d+r\d+", counts_file.as_posix()): + return "SKIP" + else: + return _process_standard_replicate(counts_file, strand_file, sample_name) + + +async def _create_sample_counts_matrix(metrics: _StudyMetrics) -> pd.DataFrame: + adjusted_index = 0 + counts: pd.DataFrame | Literal["SKIP"] = _prepare_sample_counts( + sample_name=metrics.sample_names[0], + counts_file=metrics.count_files[0], + strand_file=metrics.strandedness_files[0], + all_counts_files=metrics.count_files, + ) + + for i in range(1, metrics.num_samples): + new_counts = _prepare_sample_counts( + sample_name=metrics.sample_names[i], + counts_file=metrics.count_files[i], + strand_file=metrics.strandedness_files[i], + all_counts_files=metrics.count_files, + ) + if isinstance(new_counts, str) and new_counts == "SKIP": + adjusted_index += 1 + continue + + counts: pd.DataFrame = counts.merge(new_counts, on="ensembl_gene_id", how="outer") + counts = counts.fillna(value=0) + + # Remove run number "r\d+" from multi-run names + if re.search(r"R\d+r1", metrics.sample_names[i]): + new_sample_name = re.sub(r"r\d+", "", metrics.sample_names[i]) + counts.columns[i + 1 - adjusted_index] = new_sample_name + + return counts + + +async def _create_counts_matrix(context_name: str, config: Config): + """Create a counts matrix by reading gene counts table(s).""" + data_dir = config.data_dir / "COMO_input" / context_name matrix_output_dir = config.data_dir / "data_matrices" / context_name - print(f"Creating Counts Matrix for '{context_name}'") - # call generate_counts_matrix.R to create count matrix from COMO_input folder - rpy2_api.Rpy2( - r_file_path=r_file_path, - data_dir=input_dir.as_posix(), - out_dir=matrix_output_dir.as_posix(), - ).call_function("generate_counts_matrix_main") + study_metrics = _organize_gene_counts_files(data_dir=data_dir) + final_matrix: pd.DataFrame = pd.DataFrame() + for metric in study_metrics: + counts: pd.DataFrame = await _create_sample_counts_matrix(metric) + final_matrix = ( + counts if final_matrix.empty else pd.merge(final_matrix, counts, on="ensembl_gene_id", how="outer") + ) -def create_config_df(context_name): - """ - Create configuration sheet at /main/data/config_sheets/rnaseq_data_inputs_auto.xlsx - based on the gene counts matrix. If using zFPKM normalization technique, fetch mean fragment lengths from - /work/data/COMO_input///fragmentSizes/ + output_filename = matrix_output_dir / f"gene_counts_matrix_full_{data_dir.stem}.csv" + output_filename.parent.mkdir(parents=True, exist_ok=True) + final_matrix.to_csv(output_filename, index=False) + logger.success(f"Wrote gene count matrix for '{data_dir.stem}' at '{output_filename}'") + + +async def _create_config_df(context_name: str) -> pd.DataFrame: # noqa: C901 + """Create configuration sheet. + + The configuration file created is based on the gene counts matrix. + If using zFPKM normalization technique, mean fragment lengths will be fetched """ config = Config() gene_counts_files = list(Path(config.data_dir, "COMO_input", context_name, "geneCounts").rglob("*.tab")) - out_df = pd.DataFrame(columns=["SampleName", "FragmentLength", "Layout", "Strand", "Group"]) + sample_names: list[str] = [] + fragment_lengths: list[int | float] = [] + layouts: list[str] = [] + strands: list[str] = [] + groups: list[str] = [] + preparation_method: list[str] = [] - for gcfilename in gene_counts_files: + for gene_count_filename in sorted(gene_counts_files): try: # Match S___R___r___ # \d{1,3} matches 1-3 digits # (?:r\d{1,3})? matches an option "r" followed by three digits - label = re.findall(r"S\d{1,3}R\d{1,3}(?:r\d{1,3})?", gcfilename.as_posix())[0] + label = re.findall(r"S\d{1,3}R\d{1,3}(?:r\d{1,3})?", gene_count_filename.as_posix())[0] - except IndexError: + except IndexError as e: raise IndexError( - f"\n\nFilename of '{gcfilename}' is not valid. Should be 'contextName_SXRYrZ.tab', where X is the " - "study/batch number, Y is the replicate number, and Z is the run number." + f"\n\nFilename of '{gene_count_filename}' is not valid. " + f"Should be 'contextName_SXRYrZ.tab', where X is the study/batch number, Y is the replicate number, " + f"and Z is the run number." "\n\nIf not a multi-run sample, exclude 'rZ' from the filename." - ) + ) from e study_number = re.findall(r"S\d{1,3}", label)[0] rep_number = re.findall(r"R\d{1,3}", label)[0] @@ -74,67 +327,74 @@ def create_config_df(context_name): for r in runs: r_label = re.findall(r"r\d{1,3}", r.as_posix())[0] - R_label = re.findall(r"R\d{1,3}", r.as_posix())[0] + R_label = re.findall(r"R\d{1,3}", r.as_posix())[0] # noqa: N806 frag_filename = "".join([context_name, "_", study_number, R_label, r_label, "_fragment_size.txt"]) - frag_files.append(config.data_dir / "COMO_input" / context_name / "fragmentSizes" / study_number / frag_filename) - - layout_filename = context_name + "_" + label + "_layout.txt" - strand_filename = context_name + "_" + label + "_strandedness.txt" - frag_filename = context_name + "_" + label + "_fragment_size.txt" - prep_filename = context_name + "_" + label + "_prep_method.txt" + frag_files.append( + config.data_dir / "COMO_input" / context_name / "fragmentSizes" / study_number / frag_filename + ) context_path = config.data_dir / "COMO_input" / context_name - layout_path = context_path / "layouts" - strand_path = context_path / "strandedness" - frag_path = context_path / "fragmentSizes" - prep_path = context_path / "prepMethods" - layout_files = list(layout_path.rglob(layout_filename)) - strand_files = list(strand_path.rglob(strand_filename)) - frag_files = list(frag_path.rglob(frag_filename)) - prep_files = list(prep_path.rglob(prep_filename)) + layout_files: list[Path] = list((context_path / "layouts").rglob(f"{context_name}_{label}_layout.txt")) + strand_files: list[Path] = list( + (context_path / "strandedness").rglob(f"{context_name}_{label}_strandedness.txt") + ) + frag_files: list[Path] = list( + (context_path / "fragmentSizes").rglob(f"{context_name}_{label}_fragment_size.txt") + ) + prep_files: list[Path] = list((context_path / "prepMethods").rglob(f"{context_name}_{label}_prep_method.txt")) - # Get layout layout = "UNKNOWN" if len(layout_files) == 0: - print( - f"No layout file found for {label}, writing as 'UNKNOWN', this should be defined by user if using zFPKM or rnaseq_gen.py will not run" + logger.warning( + f"No layout file found for {label}, writing as 'UNKNOWN', " + f"this should be defined by user if using zFPKM or rnaseq_gen.py will not run" ) elif len(layout_files) == 1: - with open(layout_files[0]) as file: + with layout_files[0].open("w") as file: layout = file.read().strip() elif len(layout_files) > 1: - raise ValueError(f"Multiple matching layout files for {label}, make sure there is only one copy for each replicate in COMO_input") + raise ValueError( + f"Multiple matching layout files for {label}, " + f"make sure there is only one copy for each replicate in COMO_input" + ) - # Get strandedness strand = "UNKNOWN" if len(strand_files) == 0: - print( + logger.warning( f"No strandedness file found for {label}, writing as 'UNKNOWN'. " - f"This will not interfere with the analysis since you have already set rnaseq_preprocess.py to infer the strandedness when writing the counts matrix" + f"This will not interfere with the analysis since you have already set rnaseq_preprocess.py to " + f"infer the strandedness when writing the counts matrix" ) elif len(strand_files) == 1: - with open(strand_files[0]) as file: + with strand_files[0].open("w") as file: strand = file.read().strip() elif len(strand_files) > 1: - raise ValueError(f"Multiple matching strandedness files for {label}, make sure there is only one copy for each replicate in COMO_input") + raise ValueError( + f"Multiple matching strandedness files for {label}, " + f"make sure there is only one copy for each replicate in COMO_input" + ) - # Get preparation method prep = "total" if len(prep_files) == 0: - print(f"No prep file found for {label}, assuming 'total' as in Total RNA library preparation") + logger.warning(f"No prep file found for {label}, assuming 'total' as in Total RNA library preparation") elif len(prep_files) == 1: - with open(prep_files[0]) as file: + with prep_files[0].open("w") as file: prep = file.read().strip().lower() if prep not in ["total", "mrna"]: raise ValueError(f"Prep method must be either 'total' or 'mrna' for {label}") elif len(prep_files) > 1: - raise ValueError(f"Multiple matching prep files for {label}, make sure there is only one copy for each replicate in COMO_input") + raise ValueError( + f"Multiple matching prep files for {label}, " + f"make sure there is only one copy for each replicate in COMO_input" + ) - # Get fragment length mean_fragment_size = 100 if len(frag_files) == 0: - print(f"\nNo fragment file found for {label}, using '100'. This must be defined by the user in order to use zFPKM normalization") + logger.warning( + f"No fragment file found for {label}, using '100'. " + f"This must be defined by the user in order to use zFPKM normalization" + ) elif len(frag_files) == 1: if layout == "single-end": mean_fragment_size = 0 @@ -156,180 +416,233 @@ def create_config_df(context_name): mean_fragment_size = sum(mean_fragment_sizes * library_sizes) / sum(library_sizes) elif len(frag_files) > 1: - raise ValueError(f"Multiple matching fragment files for {label}, make sure there is only one copy for each replicate in COMO_input") - - new_row = pd.DataFrame( - { - "SampleName": [f"{context_name}_{study_number}{rep_number}"], - "FragmentLength": [mean_fragment_size], - "Layout": [layout], - "Strand": [strand], - "Group": [study_number], - "LibraryPrep": [prep], - } - ) - - out_df = pd.concat([out_df, new_row], sort=True) - out_df.sort_values("SampleName", inplace=True) + raise ValueError( + f"Multiple matching fragment files for {label}, " + f"make sure there is only one copy for each replicate in COMO_input" + ) + sample_names.append(f"{context_name}_{study_number}{rep_number}") + fragment_lengths.append(mean_fragment_size) + layouts.append(layout) + strands.append(strand) + groups.append(study_number) + preparation_method.append(prep) + + out_df = pd.DataFrame( + { + "sample_name": sample_names, + "fragment_length": fragment_lengths, + "layout": layouts, + "strand": strands, + "study": groups, + "library_prep": preparation_method, + } + ).sort_values("sample_name") return out_df -def split_config_df(df): - """ - Split a config dataframe into two seperate ones. One for Total RNA library prep, one for mRNA +def _split_config_df(df): + """Split a config dataframe to two. + + One for Total RNA library prep, one for mRNA """ - df_t = df[df["LibraryPrep"] == "total"] - df_m = df[df["LibraryPrep"] == "mrna"] + df_t = df[df["library_prep"] == "total"] + df_m = df[df["library_prep"] == "mrna"] return df_t, df_m -def split_counts_matrices(count_matrix_all, df_total, df_mrna): - """ - Split a counts-matrix dataframe into two seperate ones. One for Total RNA library prep, one for mRNA +def _split_counts_matrices( + count_matrix_all: Path, df_total: pd.DataFrame, df_mrna: pd.DataFrame +) -> tuple[pd.DataFrame, pd.DataFrame]: + """Split a counts-matrix dataframe to two. + + One for Total RNA library prep, one for mRNA """ + logger.info(f"Reading gene count matrix file at '{count_matrix_all}'") matrix_all = pd.read_csv(count_matrix_all) - matrix_total = matrix_all[["genes"] + [n for n in matrix_all.columns if n in df_total["SampleName"].tolist()]] - matrix_mrna = matrix_all[["genes"] + [n for n in matrix_all.columns if n in df_mrna["SampleName"].tolist()]] + matrix_total = matrix_all[ + ["ensembl_gene_id"] + [n for n in matrix_all.columns if n in df_total["sample_name"].tolist()] + ] + matrix_mrna = matrix_all[ + ["ensembl_gene_id"] + [n for n in matrix_all.columns if n in df_mrna["sample_name"].tolist()] + ] return matrix_total, matrix_mrna -def create_gene_info_file(matrix_file_list: list[str], input_format: Input, taxon_id): - """ - Create gene info file for specified context by reading first column in its count matrix file at - results//gene_info_.csv - """ - config = Config() - - print("Fetching gene info") - gene_info_file = config.data_dir / "gene_info.csv" - genes: list[str] = [] - for file in matrix_file_list: - genes += pd.read_csv(file)["genes"].tolist() - genes = list(set(genes)) - - # Create our output database format - # Do not include values equal to "form" - # Remove items not equal to `form` because the input database cannot exist as an output database - output_db: list[Output] = [ - i for i in [Output.ENSEMBL_GENE_ID, Output.GENE_SYMBOL, Output.GENE_ID, Output.CHROMOSOMAL_LOCATION] if i.value != input_format.value - ] - - biodbnet = BioDBNet() - gene_info = biodbnet.db2db( - input_values=genes, - input_db=input_format, - output_db=output_db, +async def _create_gene_info_file( + *, + matrix_files: list[Path], + taxon_id, + config: Config, + cache: bool, +): + """Create gene info file for specified context by reading first column in its count matrix file.""" + logger.info("Fetching gene info") + genes = set() + for file in matrix_files: + data: pd.DataFrame | sc.AnnData = pd.read_csv(file) if file.suffix == ".csv" else sc.read_h5ad(file) + input_values = data.iloc[:, 0].tolist() if isinstance(data, pd.DataFrame) else data.var_names.tolist() + conversion = await convert_gene_data(input_values, taxon_id) + genes.update(conversion["entrez_gene_id"].astype(str).tolist()) + + mygene = MyGene(cache=cache) + gene_data = await mygene.query( + items=list(genes), taxon=taxon_id, + scopes="entrezgene", ) - - gene_info.rename(columns={Output.ENSEMBL_GENE_ID.value: "ensembl_gene_id"}, inplace=True) - gene_info["start_position"] = gene_info["Chromosomal Location"].str.extract(r"chr_start: (\d+)") - gene_info["end_position"] = gene_info["Chromosomal Location"].str.extract(r"chr_end: (\d+)") - gene_info.rename(columns={"Gene Symbol": "hgnc_symbol", "Gene ID": "entrezgene_id"}, inplace=True) - gene_info.drop(["Chromosomal Location"], axis=1, inplace=True) - gene_info.to_csv(gene_info_file, index=False) - print(f"Gene Info file written at '{gene_info_file}'") - - -def handle_context_batch(context_names, mode, input_format: Input, taxon_id, provided_matrix_file): - """ - Handle iteration through each context type and create files according to flag used (config, matrix, info) - """ - config = Config() + gene_info: pd.DataFrame = pd.DataFrame( + data=None, + columns=pd.Index(data=["ensembl_gene_id", "gene_symbol", "entrez_gene_id", "start_position", "end_position"]), + index=pd.Index(data=range(len(gene_data))), + ) + for i, data in enumerate(gene_data): + ensembl_ids = data.get("ensembl.gene", "-") + if isinstance(ensembl_ids, list): + ensembl_ids = ensembl_ids[0] + + start_pos = data.get("genomic_pos.start", 0) + end_pos = data.get("genomic_pos.end", 0) + if isinstance(start_pos, list): + start_pos = sum(start_pos) / len(start_pos) + if isinstance(end_pos, list): + end_pos = sum(end_pos) / len(end_pos) + + gene_info.at[i, "gene_symbol"] = data.get("symbol", "-") + gene_info.at[i, "entrez_gene_id"] = data.get("entrezgene", "-") + gene_info.at[i, "ensembl_gene_id"] = ensembl_ids + gene_info.at[i, "start_position"] = start_pos + gene_info.at[i, "end_position"] = end_pos + + gene_info = gene_info[ + (gene_info["entrez_gene_id"] != "-") & (gene_info["ensembl_gene_id"] != "-") & (gene_info["gene_symbol"] != "-") + ] + gene_info["size"] = gene_info["end_position"].astype(int) - gene_info["start_position"].astype(int) + gene_info.drop(columns=["start_position", "end_position"], inplace=True) + output_filepath = config.data_dir / "gene_info.csv" + gene_info.to_csv(output_filepath, index=False) + logger.success(f"Gene Info file written at '{output_filepath}'") + + +async def _handle_context_batch( # noqa: C901 + context_names: list[str], + mode: Literal["create", "provide"], + taxon_id, + provided_matrix_file, + config: Config, + cache: bool, +): + """Handle iteration through each context type and create appropriate files.""" trnaseq_config_filename = config.config_dir / "trnaseq_data_inputs_auto.xlsx" mrnaseq_config_filename = config.config_dir / "mrnaseq_data_inputs_auto.xlsx" - tflag = False # turn on when any total set is found to prevent writer from being init multiple times or empty - mflag = False # turn on when any mrna set is found to prevent writer from being init multiple times or empty - - print(f"Found {len(context_names)} contexts to process: {', '.join(context_names)}") - - tmatrix_files = [] - mmatrix_files = [] - for context_name in context_names: - context_name = context_name.strip(" ") - print(f"Preprocessing {context_name}") - gene_output_dir = config.result_dir / context_name - matrix_output_dir = config.data_dir / "data_matrices" / context_name - - gene_output_dir.parent.mkdir(parents=True, exist_ok=True) - matrix_output_dir.parent.mkdir(parents=True, exist_ok=True) - - print('Gene info output directory is "{}"'.format(gene_output_dir)) - - matrix_path_all = matrix_output_dir / f"gene_counts_matrix_full_{context_name}.csv" - matrix_path_total = matrix_output_dir / f"gene_counts_matrix_total_{context_name}.csv" - matrix_path_mrna = matrix_output_dir / f"gene_counts_matrix_mrna_{context_name}.csv" - - if mode == "make": - create_counts_matrix(context_name) - # TODO: warn user or remove samples that are all 0 to prevent density plot error in zFPKM - df = create_config_df(context_name) - df_t, df_m = split_config_df(df) - - if not df_t.empty: - if not tflag: - tflag = True - twriter = pd.ExcelWriter(trnaseq_config_filename) - - tmatrix_files.append(matrix_path_total) - df_t.to_excel(twriter, sheet_name=context_name, header=True, index=False) - - if not df_m.empty: - if not mflag: - mflag = True - mwriter = pd.ExcelWriter(mrnaseq_config_filename) - - mmatrix_files.append(matrix_path_mrna) - df_m.to_excel(mwriter, sheet_name=context_name, header=True, index=False) - - tmatrix, mmatrix = split_counts_matrices(matrix_path_all, df_t, df_m) - if len(tmatrix.columns) >= 1: - tmatrix.to_csv(matrix_path_total, header=True, index=False) - if len(mmatrix.columns) >= 1: - mmatrix.to_csv(matrix_path_mrna, header=True, index=False) - - if mode == "make": - if tflag: - twriter.close() - if mflag: - mwriter.close() - - create_gene_info_file(tmatrix_files + mmatrix_files, input_format, taxon_id) - - else: - matrix_files: list[str] = como_utilities.stringlist_to_list(provided_matrix_file) - create_gene_info_file(matrix_files, input_format, taxon_id) - - -def rnaseq_preprocess(context_names: str, mode: str, input_format: Input, taxon_id: Union[int, str], matrix_file: Optional[str] = None) -> None: - if not mode == "make" and not mode == "provide": - raise ValueError("mode must be either 'make' or 'provide'") - - if input_format not in [Input.ENSEMBL_GENE_ID, Input.GENE_SYMBOL, Input.GENE_ID]: - raise ValueError("input_format must be either 'ENSEMBL_GENE_ID', 'GENE_SYMBOL', or 'GENE_ID'") + using_trna = False # turn on when any total set is found to prevent writer from being init multiple times or empty + using_mrna = False # turn on when any mrna set is found to prevent writer from being init multiple times or empty + + logger.success(f"Found {len(context_names)} contexts to process: {', '.join(context_names)}") + + tmatrix_files: list[Path] = [] + mmatrix_files: list[Path] = [] + match mode: + case "create": + for context_name in context_names: + context_name = context_name.strip(" ") + logger.info(f"Processing {context_name}") + gene_output_dir = config.result_dir / context_name + matrix_output_dir = config.data_dir / "data_matrices" / context_name + + gene_output_dir.parent.mkdir(parents=True, exist_ok=True) + matrix_output_dir.parent.mkdir(parents=True, exist_ok=True) + + logger.info(f"Gene info output directory is '{gene_output_dir}'") + + await _create_counts_matrix(context_name, config=config) + # TODO: warn user or remove samples that are all 0 to prevent density plot error in zFPKM + config_df = await _create_config_df(context_name) + trna_df, mrna_df = _split_config_df(config_df) + + matrix_path_total = matrix_output_dir / f"gene_counts_matrix_total_{context_name}.csv" + if not trna_df.empty: + if not using_trna: + using_trna = True + twriter = pd.ExcelWriter(trnaseq_config_filename) + + tmatrix_files.append(matrix_path_total) + trna_df.to_excel(twriter, sheet_name=context_name, header=True, index=False) + + matrix_path_mrna = matrix_output_dir / f"gene_counts_matrix_mrna_{context_name}.csv" + if not mrna_df.empty: + if not using_mrna: + using_mrna = True + mwriter = pd.ExcelWriter(mrnaseq_config_filename) + + mmatrix_files.append(matrix_path_mrna) + mrna_df.to_excel(mwriter, sheet_name=context_name, header=True, index=False) + + matrix_path_all = matrix_output_dir / f"gene_counts_matrix_full_{context_name}.csv" + trna_matrix, mrna_matrix = _split_counts_matrices(matrix_path_all, trna_df, mrna_df) + if len(trna_matrix.columns) >= 1: + trna_matrix.to_csv(matrix_path_total, header=True, index=False) + if len(mrna_matrix.columns) >= 1: + mrna_matrix.to_csv(matrix_path_mrna, header=True, index=False) + + if using_trna: + twriter.close() + if using_mrna: + mwriter.close() + + await _create_gene_info_file( + matrix_files=tmatrix_files + mmatrix_files, + taxon_id=taxon_id, + config=config, + cache=cache, + ) + case "provide": + matrix_files: list[Path] = [Path(p) for p in stringlist_to_list(provided_matrix_file)] + await _create_gene_info_file( + matrix_files=matrix_files, + taxon_id=taxon_id, + config=config, + cache=cache, + ) + case _: + raise ValueError("'--mode' must be either 'create' or 'provide'") + + +async def rnaseq_preprocess( + context_names: list[str], + mode: Literal["create", "provide"], + taxon_id: int | str, + input_format: Input | str, + matrix_file: str | Path | None = None, + config: Config = None, + cache: bool = True, +) -> None: + """Preprocesses RNA-seq data for downstream analysis. + + Fetches additional gene information from a provided matrix or gene counts, + or optionally creates this matrix using gene count files obtained using STAR aligner + """ + config = Config() if config is None else config + if mode not in {"create", "provide"}: + raise ValueError("mode must be either 'create' or 'provide'") if not isinstance(taxon_id, int) and taxon_id not in ["human", "mouse"]: raise ValueError("taxon_id must be either an integer, or accepted string ('mouse', 'human')") - handle_context_batch(context_names=context_names, mode=mode, input_format=input_format, taxon_id=taxon_id, provided_matrix_file=matrix_file) - - -def parse_args(): - """ - Parse arguments to rnaseq_preprocess.py, create a gene info files for each provided context at: - /work/data/results//gene_info_.csv. - - If using --info-matrix or --info-matrix-config: - create gene count matrix file at /work/data/data_matrices//gene_counts_matrix_.csv, + await _handle_context_batch( + context_names=context_names, + mode=mode, + taxon_id=taxon_id, + provided_matrix_file=Path(matrix_file if matrix_file is not None else "").as_posix(), + config=config, + cache=cache, + ) - If using --info-matrix-config: - create config file at /work/data/config_sheets/rnaseq_data_inputs_auto.xlsx - """ +def _parse_args(): parser = argparse.ArgumentParser( prog="rnaseq_preprocess.py", description=""" @@ -343,7 +656,6 @@ def parse_args(): For additional help, please post questions/issues in the MADRID GitHub repo at https://github.com/HelikarLab/MADRID or email babessell@gmail.com""", ) - parser.add_argument( "-n", "--context-names", @@ -357,107 +669,73 @@ def parse_args(): the count matrix as an imported .csv file. If making multiple models in a batch, then use the format: "context1 context2 context3". """, ) - parser.add_argument( - "-f", - "--gene-format", - type=str, + "-i", + "--taxon-id", required=False, - default="Ensembl Gene ID", - dest="gene_format", - help="Format of Genes, accepts 'Ensembl', 'Entrez', or'HGNC symbol'", + default=9606, + dest="taxon_id", + help="BioDbNet taxon ID number, also accepts 'human', or 'mouse'", ) - parser.add_argument( - "-i", "--taxon-id", required=False, default=9606, dest="taxon_id", help="BioDbNet taxon ID number, also accepts 'human', or 'mouse'" + "--input-format", + required=True, + dest="input_format", + help="The data input format, such as Ensembl, Entrez, etc.", ) - - group = parser.add_mutually_exclusive_group(required=True) - - group.add_argument( - "-p", - "--provide-matrix", - action="store_true", - required=False, - default=False, - dest="provide_matrix", - help="Provide your own count matrix. Requires additional argument '--matrix' which is .csv file " - "where colnames are sample names (in contextName_SXRY format) and rownames are genes in " - "in format specified by --gene-format", - ) # would be nice if this was a directory full matrices in case you want to do in batches - - group.add_argument( - "-y", - "--create-matrix", - action="store_true", - required=False, - default=False, - dest="make_matrix", - help="Flag for if you want to make a counts matrix, but not a config file. " - "Requires a correctly structured COMO_input folder in /work/data/. Can make one using: " - "https://github.com/HelikarLab/FastqToGeneCounts", + parser.add_argument( + "--mode", + type=str, + required=True, + dest="mode", + choices={"create", "provide"}, + help="Mode of rnaseq_preprocess.py, either 'make' or 'provide'", ) - parser.add_argument( - "-m", "--matrix", - required="--provide-matrix" in sys.argv, # require if using --provide-matrix flag, + required=False, dest="provided_matrix_fname", - default="SKIP", - help="Name of provided counts matrix in " "/work/data/data_matrices//.csv", + default=None, + help="Name of provided counts matrix in /work/data/data_matrices//.csv", ) - args = parser.parse_args() - args.context_names = como_utilities.stringlist_to_list(args.context_names) - - return args - - -def main(): - args = parse_args() - - if args.gene_format.upper() in ["ENSEMBL", "ENSEMBLE", "ENSG", "ENSMUSG", "ENSEMBL ID", "ENSEMBL GENE ID"]: - gene_format_database: Input = Input.ENSEMBL_GENE_ID - - elif args.gene_format.upper() in ["HGNC SYMBOL", "HUGO", "HUGO SYMBOL", "SYMBOL", "HGNC", "GENE SYMBOL"]: - gene_format_database: Input = Input.GENE_SYMBOL + parsed = parser.parse_args() + parsed.context_names = stringlist_to_list(parsed.context_names) - elif args.gene_format.upper() in ["ENTREZ", "ENTRES", "ENTREZ ID", "ENTREZ NUMBER" "GENE ID"]: - gene_format_database: Input = Input.GENE_ID - - else: # provided invalid gene format - raise ValueError(f"Gene format (--gene_format) is invalid; accepts 'Ensembl', 'Entrez', and 'HGNC symbol'; provided: {args.gene_format}") + if parsed.mode == "provide" and parsed.provided_matrix_fname is None: + raise ValueError("If provide_matrix is True, then provided_matrix_fname must be provided") # handle species alternative ids - if isinstance(args.taxon_id, str): - if args.taxon_id.upper() == "HUMAN" or args.taxon_id.upper() == "HOMO SAPIENS": - taxon_id = Taxon.HOMO_SAPIENS - elif args.taxon_id.upper() == "MOUSE" or args.taxon_id.upper() == "MUS MUSCULUS": - taxon_id = Taxon.MUS_MUSCULUS - else: - raise ValueError(f"Taxon id (--taxon-id) is invalid; accepts 'human', 'mouse'; provided: {args.taxon_id}") - - elif isinstance(args.taxon_id, int): - taxon_id = args.taxon_id - else: - raise ValueError(f"Taxon id (--taxon-id) is invalid; accepts 'human', 'mouse'; provided: {args.taxon_id}") - - # use mutually exclusive flag to set mode which tells which files to generate - if args.provide_matrix: - mode = "provide" - elif args.make_matrix: - mode = "make" + taxon_id = str(parsed.taxon_id) + if taxon_id.isdigit(): + try: + parsed.taxon_id = Taxon.from_int(int(taxon_id)) + except ValueError: + parsed.taxon_id = int(taxon_id) else: - raise ValueError("Must set either --provide-matrix or --make-matrix") + if taxon_id.upper() in {"HUMAN", "HOMO SAPIENS"}: + parsed.taxon_id = Taxon.HOMO_SAPIENS + elif taxon_id.upper() in {"MOUSE", "MUS MUSCULUS"}: + parsed.taxon_id = Taxon.MUS_MUSCULUS + else: + raise ValueError( + f"Taxon id (--taxon-id) is invalid; accepts 'human', 'mouse', or an integer value; provided: {taxon_id}" + ) - handle_context_batch( - context_names=args.context_names, - mode=mode, - input_format=gene_format_database, - taxon_id=taxon_id, - provided_matrix_file=args.provided_matrix_fname, - ) + args = _Arguments(**vars(parsed)) + return args if __name__ == "__main__": - main() + args: _Arguments = _parse_args() + taxon_id_value = args.taxon_id.value if isinstance(args.taxon_id, Taxon) else args.taxon_id + + asyncio.run( + rnaseq_preprocess( + context_names=args.context_names, + mode=args.mode, + taxon_id=taxon_id_value, + matrix_file=args.provided_matrix_fname, + input_format=args.input_format, + ) + ) diff --git a/main/como/rpy2_api.py b/main/como/rpy2_api.py index 4d182162..910552ec 100644 --- a/main/como/rpy2_api.py +++ b/main/como/rpy2_api.py @@ -1,3 +1,5 @@ +# ruff: noqa + import typing from pathlib import Path @@ -24,7 +26,7 @@ def call_function(self, r_function_name: str, *args, **kwargs) -> typing.Any: self._args = args if self._kwargs == {}: self._kwargs = kwargs - + func_ = rpy2.robjects.packages.SignatureTranslatedAnonymousPackage(self._r_file_read, "func_") # Dynamically call the func_ function, using the arguments passed in diff --git a/main/como/rscripts/DGE.R b/main/como/rscripts/DGE.R index a6634629..341a2553 100644 --- a/main/como/rscripts/DGE.R +++ b/main/como/rscripts/DGE.R @@ -1,17 +1,18 @@ suppressPackageStartupMessages(library("DESeq2")) suppressPackageStartupMessages(library("edgeR")) +suppressPackageStartupMessages(library("readr")) suppressPackageStartupMessages(library("readxl")) suppressPackageStartupMessages(library("stringr")) work_dir <- getwd() -r_log_directory <- str_interp("${work_dir}/logs") +r_log_directory <- stringr::str_interp("${work_dir}/logs") if (!dir.exists(r_log_directory)) { dir.create(r_log_directory) } zz <- file(file.path(r_log_directory, "DGE.Rout"), open = "wt") sink(zz, type = "message") readCountMatrix <- function(cmat_file, config_file, disease_name) { - conf <- read_excel(config_file, sheet = disease_name, col_names = TRUE) - cmat_whole <- read.csv(cmat_file, header = TRUE) + conf <- readxl::read_excel(config_file, sheet = disease_name, col_names = TRUE) + cmat_whole <- readr::read_csv(cmat_file) cmat_whole[, -1] <- lapply(cmat_whole[, -1], as.numeric) cmat_whole <- cmat_whole[rowSums(cmat_whole[, -1]) > 0,] genes <- as.character(cmat_whole$genes) diff --git a/main/como/rscripts/combine_distributions.R b/main/como/rscripts/combine_distributions.R index a1fa55d0..54a888dd 100644 --- a/main/como/rscripts/combine_distributions.R +++ b/main/como/rscripts/combine_distributions.R @@ -6,7 +6,7 @@ suppressPackageStartupMessages(library("ggplot2")) # Check if rlogs directory exists, From: https://stackoverflow.com/a/46008094 # Then prevent messy messages from repeatedly writing to juypter work_dir <- getwd() -r_log_directory <- str_interp("${work_dir}/logs") +r_log_directory <- stringr::str_interp("${work_dir}/logs") if (!dir.exists(r_log_directory)) { dir.create(r_log_directory) } zz <- file(file.path(r_log_directory, "combine_distributions.Rout"), open = "wt") sink(zz, type = "message") @@ -427,213 +427,195 @@ combine_omics_zdistros <- function( combine_zscores_main <- function( - wd, - contexts, - use_mrna_flag, - use_trna_flag, - use_scrna_flag, - use_proteins_flag, + working_dir, + context_names, + global_use_mrna, + global_use_trna, + global_use_scrna, + global_use_proteins, keep_gene_scores, - tweight_master = 1, - mweight_master = 1, - sweight_master = 1, - pweight_master = 2 + global_trna_weight, + global_mrna_weight, + global_scrna_weight, + global_protein_weight ) { - fig_path <- file.path(wd, "figures") - if (!file.exists(fig_path)) { dir.create(fig_path) } + figure_output_dir <- file.path(working_dir, "figures") + if (!file.exists(figure_output_dir)) { dir.create(figure_output_dir) } - tbatches <- parse_contexts_zfpkm(wd, contexts, "total") - mbatches <- parse_contexts_zfpkm(wd, contexts, "mrna") - sbatches <- parse_contexts_zumi(wd, contexts, "scrna") - pbatches <- parse_contexts_zscore_prot(wd, contexts) + global_trna_batches <- parse_contexts_zfpkm(working_dir, context_names, "total") + global_mrna_batches <- parse_contexts_zfpkm(working_dir, context_names, "mrna") + global_scrna_batches <- parse_contexts_zumi(working_dir, context_names, "scrna") + global_protein_batches <- parse_contexts_zscore_prot(working_dir, context_names) - for (context in contexts) { - use_mrna <- use_mrna_flag - use_trna <- use_trna_flag - use_scrna <- use_scrna_flag - use_proteins <- use_proteins_flag + for (context in context_names) { + context_use_trna <- global_use_trna + context_use_mrna <- global_use_mrna + context_use_scrna <- global_use_scrna + context_use_proteins <- global_use_proteins - tweight <- tweight_master - mweight <- mweight_master - sweight <- sweight_master - pweight <- pweight_master + context_trna_weight <- global_trna_weight + context_mrna_weight <- global_mrna_weight + context_scrna_weight <- global_scrna_weight + context_protein_weight <- global_protein_weight - cont_tbatches <- tbatches[[context]] - cont_mbatches <- mbatches[[context]] - cont_sbatches <- sbatches[[context]] - cont_pbatches <- pbatches[[context]] + context_trna_batch <- global_trna_batches[[context]] + context_mrna_batch <- global_mrna_batches[[context]] + context_scrna_batch <- global_scrna_batches[[context]] + context_protein_batch <- global_protein_batches[[context]] - if (length(cont_tbatches) == 0 & use_trna_flag) { - use_trna <- FALSE + if (length(context_trna_batch) == 0 & global_use_trna) { + context_use_trna <- FALSE print(paste0("No total RNA-seq zFPKM Matrix files found for ", context, ". Will not use for this context.")) } - if (length(cont_mbatches) == 0 & use_mrna_flag) { - use_mrna <- FALSE + if (length(context_mrna_batch) == 0 & global_use_mrna) { + context_use_mrna <- FALSE print(paste0("No polyA RNA-seq zFPKM Matrix files found for ", context, ". Will not use for this context.")) } - if (length(cont_sbatches) == 0 & use_scrna_flag) { - use_scrna <- FALSE + if (length(context_scrna_batch) == 0 & global_use_scrna) { + context_use_scrna <- FALSE print(paste0("No SC RNA-seq zFPKM Matrix files found for ", context, ". Will not use for this context.")) } - if (length(cont_pbatches) == 0 & use_proteins_flag) { - use_proteins <- FALSE + if (length(context_protein_batch) == 0 & global_use_proteins) { + context_use_proteins <- FALSE print(paste0("No proteomics z-score Matrix files found for ", context, ". Will not use for this context.")) } - if (use_trna) { + if (context_use_trna) { print("Will merge total RNA-seq distributions") - twd <- file.path(wd, context, "total") - nreps <- c() - cnt <- 0 - for (batch in cont_tbatches) { - res <- merge_batch(twd, context, batch) + trna_workdir <- file.path(working_dir, context, "total") + num_reps <- c() + count <- 0 + for (batch in context_trna_batch) { + res <- merge_batch(trna_workdir, context, batch) zmat <- res[[1]] - nreps <- c(nreps, res[[2]]) - comb_z <- combine_batch_zdistro(twd, context, batch, zmat) + num_reps <- c(num_reps, res[[2]]) + comb_z <- combine_batch_zdistro(trna_workdir, context, batch, zmat) colnames(comb_z) <- c("ENTREZ_GENE_ID", batch) - if (!cnt) { merge_z <- comb_z } + if (!count) { merge_z <- comb_z } else { merge_z <- full_join(merge_z, comb_z, by = "ENTREZ_GENE_ID") } - cnt <- cnt + 1 + count <- count + 1 } - comb_batches_z_trna <- combine_context_zdistro(twd, context, nreps, merge_z) - filename <- file.path(twd, paste0("combined_zFPKM_", context, ".csv")) + comb_batches_z_trna <- combine_context_zdistro(trna_workdir, context, num_reps, merge_z) + filename <- file.path(trna_workdir, paste0("combined_zFPKM_", context, ".csv")) write.csv(comb_batches_z_trna, filename, row.names = FALSE) - if (!use_proteins & !use_mrna & !use_scrna) { - filename <- file.path(wd, context, "total", paste0("model_scores_", context, ".csv")) + if (!context_use_proteins & !context_use_mrna & !context_use_scrna) { + filename <- file.path(working_dir, context, "total", paste0("model_scores_", context, ".csv")) write.csv(comb_batches_z_trna, filename, row.names = FALSE) } } else { comb_batches_z_trna <- NA } - if (use_mrna) { + if (context_use_mrna) { print("Will merge polyA enriched RNA-seq distributions") - mwd <- file.path(wd, context, "mrna") - nreps <- c() - cnt <- 0 - for (batch in cont_mbatches) { - res <- merge_batch(mwd, context, batch) + mrna_workdir <- file.path(working_dir, context, "mrna") + num_reps <- c() + count <- 0 + for (batch in context_mrna_batch) { + res <- merge_batch(mrna_workdir, context, batch) zmat <- res[[1]] - nreps <- c(nreps, res[[2]]) - comb_z <- combine_batch_zdistro(mwd, context, batch, zmat) + num_reps <- c(num_reps, res[[2]]) + comb_z <- combine_batch_zdistro(mrna_workdir, context, batch, zmat) colnames(comb_z) <- c("ENTREZ_GENE_ID", batch) - if (!cnt) { merge_z <- comb_z } + if (!count) { merge_z <- comb_z } else { merge_z <- full_join(merge_z, comb_z, by = "ENTREZ_GENE_ID") } - cnt <- cnt + 1 + count <- count + 1 } - comb_batches_z_mrna <- combine_context_zdistro(mwd, context, nreps, merge_z) - filename <- file.path(mwd, paste0("combined_zFPKM_", context, ".csv")) + comb_batches_z_mrna <- combine_context_zdistro(mrna_workdir, context, num_reps, merge_z) + filename <- file.path(mrna_workdir, paste0("combined_zFPKM_", context, ".csv")) write.csv(comb_batches_z_mrna, filename, row.names = FALSE) - if (!use_proteins & !use_trna & !use_scrna) { - filename <- file.path(mwd, paste0("model_scores_", context, ".csv")) + if (!context_use_proteins & !context_use_trna & !context_use_scrna) { + filename <- file.path(mrna_workdir, paste0("model_scores_", context, ".csv")) write.csv(comb_batches_z_mrna, filename, row.names = FALSE) } } else { comb_batches_z_mrna <- NA } - if (use_scrna) { + if (context_use_scrna) { print("Will merge single-cell RNA-seq distributions") - swd <- file.path(wd, context, "scrna") - nreps <- c() - cnt <- 0 - for (batch in cont_sbatches) { - res <- merge_batch(swd, context, batch) + scrna_workdir <- file.path(working_dir, context, "scrna") + num_reps <- c() + count <- 0 + for (batch in context_scrna_batch) { + res <- merge_batch(scrna_workdir, context, batch) zmat <- res[[1]] - nreps <- c(nreps, res[[2]]) - comb_z <- combine_batch_zdistro(swd, context, batch, zmat) + num_reps <- c(num_reps, res[[2]]) + comb_z <- combine_batch_zdistro(scrna_workdir, context, batch, zmat) colnames(comb_z) <- c("ENTREZ_GENE_ID", batch) - if (!cnt) { merge_z <- comb_z } + if (!count) { merge_z <- comb_z } else { merge_z <- full_join(merge_z, comb_z, by = "ENTREZ_GENE_ID") } - cnt <- cnt + 1 + count <- count + 1 } - comb_batches_z_scrna <- combine_context_zdistro(swd, context, nreps, merge_z) - filename <- file.path(swd, paste0("combined_zFPKM_", context, ".csv")) + comb_batches_z_scrna <- combine_context_zdistro(scrna_workdir, context, num_reps, merge_z) + filename <- file.path(scrna_workdir, paste0("combined_zFPKM_", context, ".csv")) write.csv(comb_batches_z_scrna, filename, row.names = FALSE) - if (!use_proteins & !use_trna & !use_mrna) { - filename <- file.path(swd, paste0("model_scores_", context, ".csv")) + if (!context_use_proteins & !context_use_trna & !context_use_mrna) { + filename <- file.path(scrna_workdir, paste0("model_scores_", context, ".csv")) write.csv(comb_batches_z_scrna, filename, row.names = FALSE) } } else { comb_batches_z_scrna <- NA } - if (use_proteins) { + if (context_use_proteins) { print("Will merge protein abundance distributions") - pwd = file.path(wd, context, "proteomics") - nreps <- c() - cnt <- 0 - for (batch in cont_pbatches) { - res <- merge_batch(pwd, context, batch) + protein_workdir <- file.path(working_dir, context, "proteomics") + num_reps <- c() + count <- 0 + for (batch in context_protein_batch) { + res <- merge_batch(protein_workdir, context, batch) zmat <- res[[1]] - nreps <- c(nreps, res[[2]]) - comb_z <- combine_batch_zdistro(pwd, context, batch, zmat) + num_reps <- c(num_reps, res[[2]]) + comb_z <- combine_batch_zdistro(protein_workdir, context, batch, zmat) colnames(comb_z) <- c("ENTREZ_GENE_ID", batch) - if (!cnt) { merge_z <- comb_z } + if (!count) { merge_z <- comb_z } else { merge_z <- full_join(merge_z, comb_z, by = "ENTREZ_GENE_ID") } - cnt <- cnt + 1 + count <- count + 1 } - comb_batches_z_prot <- combine_context_zdistro(pwd, context, nreps, merge_z) - filename <- file.path(pwd, paste0("combined_zscore_proteinAbundance_", context, ".csv")) + comb_batches_z_prot <- combine_context_zdistro(protein_workdir, context, num_reps, merge_z) + filename <- file.path(protein_workdir, paste0("combined_zscore_proteinAbundance_", context, ".csv")) write.csv(comb_batches_z_prot, filename, row.names = FALSE) - if (!use_mrna & !use_trna & !use_scrna) { - filename <- file.path(pwd, paste0("model_scores_", context, ".csv")) + if (!context_use_mrna & !context_use_trna & !context_use_scrna) { + filename <- file.path(protein_workdir, paste0("model_scores_", context, ".csv")) write.csv(comb_batches_z_prot, filename, row.names = FALSE) } } else { comb_batches_z_prot <- NA } - if (!use_trna) { tweight <- 0 } - if (!use_mrna) { mweight <- 0 } - if (!use_scrna) { sweight <- 0 } - if (!use_proteins) { pweight <- 0 } + if (!context_use_trna) { context_trna_weight <- 0 } + if (!context_use_mrna) { context_mrna_weight <- 0 } + if (!context_use_scrna) { context_scrna_weight <- 0 } + if (!context_use_proteins) { context_protein_weight <- 0 } comb_omics_z <- combine_omics_zdistros( - wd, + working_dir, context, comb_batches_z_trna, comb_batches_z_mrna, comb_batches_z_scrna, comb_batches_z_prot, - tweight, - mweight, - sweight, - pweight + context_trna_weight, + context_mrna_weight, + context_scrna_weight, + context_protein_weight ) - filename <- file.path(wd, context, paste0("model_scores_", context, ".csv")) + filename <- file.path(working_dir, context, paste0("model_scores_", context, ".csv")) write.csv(comb_omics_z, filename, row.names = FALSE) } -} - - -print(get_batch_name("test.csv")) - -# Call "combine_zscores_main" -# combine_zscores_main( -# wd = "/Users/joshl/PycharmProjects/MADRID/main/data/results", -# context = c("naiveB"), -# use_trna = TRUE, -# use_mrna = TRUE, -# use_scrna = FALSE, -# use_proteins = FALSE, -# keep_gene_scores = TRUE, -# tweight = 1, -# mweight = 1, -# sweight = 1, -# pweight = 2 -# ) +} \ No newline at end of file diff --git a/main/como/rscripts/generate_counts_matrix.R b/main/como/rscripts/generate_counts_matrix.R deleted file mode 100644 index 08a60ae0..00000000 --- a/main/como/rscripts/generate_counts_matrix.R +++ /dev/null @@ -1,239 +0,0 @@ -suppressPackageStartupMessages(library("tidyverse")) - -# Check if rlogs directory exists, From: https://stackoverflow.com/a/46008094 -# Then prevent messy messages from repeatedly writing to juypter -work_dir <- getwd() -r_log_directory <- str_interp("${work_dir}/logs") -if (!dir.exists(r_log_directory)) { dir.create(r_log_directory) } -zz <- file(file.path(r_log_directory, "generate_counts_matrix.Rout"), open = "wt") -sink(zz, type = "message") - -# fetch and organize COMO_input -organize_gene_counts_files <- function(data_dir) { - # fetch and organize COMO_input - # accepts path to data directory, normalization technique - - study_metrics <- list() # list storing all the samples - tissue_name <- basename(data_dir) # get tissue name from directory name - - # collect paths to files - count_dir_i <- file.path(data_dir, "geneCounts") # COMO_input/tissueName/geneCounts - count_dir <- list.dirs(path = count_dir_i, full.names = TRUE, recursive = FALSE) %>% - Filter(function(x) !any(grepl(".ipynb_checkpoints", x)), .) # COMO_input/tissueName/geneCounts/SX - - strand_dir_i <- file.path(data_dir, "strandedness") # COMO_input/tissueName/strandedness - strand_dir <- list.dirs(path = strand_dir_i, full.names = TRUE, recursive = FALSE) # tissueName/strandedness/SX - - # for each study, collect gene count files, frag files, insert size files, layouts, and strand info - for (j in 1:length(count_dir)) { - cnt_d <- count_dir[j] # study's gene count folder - sname <- basename(cnt_d) # get study name from directory name - entry <- list() # list specific to a study, top level of SampleMetrics - str_d <- file.path(data_dir, "strandedness", sname) # strandedness file directory - - #counts_glob <- paste(c(cnt_d, "/*.tab"), collapse="") - counts_glob <- file.path(cnt_d, "*.tab") - counts_files <- Sys.glob(counts_glob) # outputs of STAR - - n_replicates <- length(counts_files) # number of replicates for this study - replicate_names <- rep(0, n_replicates) # initialize empty array to store rep names - strandedness_files <- rep(0, n_replicates) # initialize empty array to store - - for (i in 1:n_replicates) { # for each replicate, get name of replicate ex. S1R1 - replicate_file <- str_match(counts_files[i], "geneCounts/\\s*(.*?)\\s*.tab")[, 2] # file path - replicate_names[i] <- basename(replicate_file) # file name only - rname <- sub(pattern = "(.*)\\..*$", replacement = "\\1", basename(replicate_file)) # rem file extension - strandedness_file <- paste0(rname, "_strandedness.txt") # use rep name to get matching strand - strandedness_files[i] <- file.path(str_d, strandedness_file) # path to strand files - - } - - entry[["CountFiles"]] <- counts_files # assign gene count file paths to study entry - entry[["NumSamples"]] <- n_replicates # assign number of replicates to study entry - entry[["SampleNames"]] <- replicate_names # assign sample names to study entry - entry[["StrandFiles"]] <- strandedness_files # assign strandedness files - study_metrics[[sname]] <- entry # assign study entry to the list of studies - - } - - return(study_metrics) -} - - -prepare_sample_counts <- function(counts_file, counts_files, strand_file) { - - # prepare a gene count file to be added to the count matrix - if (grepl("R\\d+r1", counts_file, ignore.case = FALSE)) { # if first run in a multirun sample - rm_run_number <- unlist(strsplit(counts_file, "r1"))[1] # remove run number from sample id - #split_directory <- unlist(strsplit(rm_run_number, "/")) # split from path - #basename <- split_directory[length(split_directory)] # get sample id - bname <- basename(rm_run_number) - search_str <- paste0(bname, "r\\d+") - run_files <- counts_files[grepl(search_str, counts_files)] # search for other files that are part of same run - samp_count <- NULL - - - for (f in run_files) { # for each run associated with this replicate, - run_count <- read.delim(f) # read gene counts - run_len <- length(run_count[, 1]) # number of genes - run_count <- run_count[4:run_len,] # only count mapped genes - run_count <- na.omit(run_count) # omit na if applicable - genes <- run_count[, 1] # list of genes - - strand_read <- gsub("[\r\n]", "", readChar(strand_file, file.info(strand_file)$size)) - - stopifnot((strand_read == "NONE") || - (strand_read == "SECOND_READ_TRANSCRIPTION_STRAND") || - (strand_read == "FIRST_READ_TRANSCRIPTION_STRAND") - ) - - if (strand_read == "FIRST_READ_TRANSCRIPTION_STRAND") { # forward - cl <- 3 - } else if (strand_read == "SECOND_READ_TRANSCRIPTION_STRAND") { # reverse - cl <- 4 - } else { # unstranded - cl <- 2 - } - - #cl <- which.max(colSums(run_count[,2:4])) + 1 # max counts is most likely the correct strandedness - - run_count <- data.frame(cbind(run_count[, 1], run_count[, cl])) # get counts from correct strandedness - run_count[, 1] <- genes # force rewrite genes bc rpy2 does not handle previous step properly - colnames(run_count) <- c("genes", "counts") # rename columns to descriptive headers - - if (is.null(samp_count)) { # if haven't declared sample counts yet - samp_count <- run_count # assign samp_counts to this run count if first one - } - - else { # otherwise merge - samp_count <- merge(samp_count, run_count, by = "genes", all = TRUE) - samp_count[is.na(samp_count)] <- 0 - } - } - - genes <- samp_count["genes"] - samp_count["genes"] <- NULL - samp_count <- sapply(samp_count, as.numeric) - samp_sum <- rowSums(samp_count) # sum the counts from each run - samp_count <- data.frame(cbind(genes, samp_sum)) - samp_count[, 1] <- genes # rewrite bc rpy2 is weird - colnames(samp_count)[2] <- "counts" - - return(samp_count) - } - - else if (grepl("R\\d+r", counts_file) == TRUE) { # multirun files handled when the first run is iterated on, skip - return("skip") - } - - else { # if not multirun, handle normally - samp_count <- read.delim(counts_file) # read file - samp_len <- length(samp_count[, 1]) # number of genes - samp_count <- samp_count[4:samp_len,] # only count mapped genes - samp_count <- na.omit(samp_count) # remove NAs - genes <- samp_count[, 1] # get gene names - - strand_read <- gsub("[\r\n]", "", readChar(strand_file, file.info(strand_file)$size)) - - stopifnot((strand_read == "NONE") || - (strand_read == "SECOND_READ_TRANSCRIPTION_STRAND") || - (strand_read == "FIRST_READ_TRANSCRIPTION_STRAND") - ) - - if (strand_read == "FIRST_READ_TRANSCRIPTION_STRAND") { # forward - cl <- 3 - } else if (strand_read == "SECOND_READ_TRANSCRIPTION_STRAND") { # reverse - cl <- 4 - } else { # unstranded - cl <- 2 - } - #cl <- which.max(colSums(samp_count[,2:4])) + 1 # max counts is most likely the correct strandedness - - samp_count <- data.frame(cbind(samp_count[, 1], samp_count[, cl])) # df from gene names and correct counts - samp_count[, 1] <- genes # force rewrite genes bc rpy2 does not handle previous step properly - colnames(samp_count) <- c("genes", "counts") # rename columns for merging - - return(samp_count) - } -} - - -create_counts_matrix <- function(counts_files, replicate_names, n_replicates, strand_files) { - # If the length of n_replicates is 1, then all samples are replicates of the same study - # Show a warning and exit the script; we cannot have less than 2 replicates - # Show the replicate name in the warning - if (n_replicates == 1) { - warning("There is only one replicate for study ", replicate_names[1], ". Please provide at least two replicates.") - stop() - } - - i_adjust <- 0 # adjusted index, subtracts number of multiruns processed - counts <- prepare_sample_counts(counts_files[1], counts_files, strand_files[1]) # get first column of counts to add - - if (grepl("R\\d+r1", replicate_names[1], ignore.case = FALSE)) { # if first of a set of multiruns - colnames(counts)[2] <- unlist(strsplit(replicate_names[1], "r\\d+"))[1] # remove run number tag - - } else { - colnames(counts)[2] <- replicate_names[1] # if not multirun, header is okay - } - - - for (i in 2:n_replicates) { # iterate through rest of replicates after handling the first and initializing matrix - new_cnts <- prepare_sample_counts(counts_files[i], counts_files, strand_files[i]) # get next column of counts to add - options(warn = -1) # turn off warnings - - if (is.character(new_cnts) && new_cnts == "skip") { # handle skipped multirun calls - i_adjust <- i_adjust + 1 - next - } - - counts <- merge(counts, new_cnts, by = "genes", all = TRUE) # merge new count list with matrix - counts[is.na(counts)] <- 0 # if gene lists differ (could be the case if aligment platforms differ) make na's 0 - - if (grepl("R\\d+r1", replicate_names[i], ignore.case = FALSE)) { # remove run number from multirun name - samp_name <- unlist(strsplit(replicate_names[i], "r\\d+"))[1] - } - else { - samp_name <- replicate_names[i] # if not multirun, sample name is okay - } - - colnames(counts)[i + 1 - i_adjust] <- samp_name # set column name of added counts - - } - - return(counts) -} - - -generate_counts_matrix_main <- function(data_dir, out_dir) { - - print("Organizing Files") - study_metrics <- organize_gene_counts_files(data_dir) - print("Creating counts matrix") - - for (i in 1:length(study_metrics)) { # for each study - res <- create_counts_matrix(study_metrics[[i]][["CountFiles"]], - study_metrics[[i]][["SampleNames"]], - study_metrics[[i]][["NumSamples"]], - study_metrics[[i]][["StrandFiles"]] - ) # create count matrix for study - - study_metrics[[i]][["CountMatrix"]] <- res # store count matrix - - study_metrics[[i]][["NumSamples"]] <- ncol(study_metrics[[i]][["CountMatrix"]]) # store number of samples - - if (i == 1) { # if first study - full_count_matrix <- study_metrics[[i]][["CountMatrix"]] # initialize supermatrix - } else { # for successive studies - add_mat <- study_metrics[[i]][["CountMatrix"]] - full_count_matrix <- merge(full_count_matrix, add_mat, by = "genes", all = TRUE) # merge study matrix to super - } - - } - - full_count_matrix["genes"] <- sub("\\.\\d+", "", full_count_matrix$genes) - file_split <- unlist(strsplit(data_dir, "/")) - file_name <- file.path(out_dir, paste0("gene_counts_matrix_full_", file_split[length(file_split)], ".csv")) - write.csv(full_count_matrix, file_name, row.names = FALSE) - cat("Count Matrix written at ", file_name, "\n") -} diff --git a/main/como/rscripts/protein_transform.R b/main/como/rscripts/protein_transform.R index 1615a78f..ce98b5e3 100644 --- a/main/como/rscripts/protein_transform.R +++ b/main/como/rscripts/protein_transform.R @@ -1,4 +1,5 @@ suppressPackageStartupMessages(library("ggplot2")) +suppressPackageStartupMessages(library("readr")) suppressPackageStartupMessages(library("tidyverse")) suppressPackageStartupMessages(library("zoo")) @@ -171,7 +172,7 @@ rm_infinite <- function(abundance) { protein_transform_main <- function(abundance_matrix, out_dir, group_name) { dir.create(file.path(out_dir, "figures"), showWarnings = FALSE) - prot <- as.data.frame(read.csv(abundance_matrix)) + prot <- as.data.frame(readr::read_csv(abundance_matrix)) prot[is.na(prot)] <- 0 min_thresh <- min(prot>0) #row.names(prot) <- prot$ENTREZ_GENE_ID diff --git a/main/como/rscripts/rnaseq.R b/main/como/rscripts/rnaseq.R index bf5146d9..23b0a37b 100644 --- a/main/como/rscripts/rnaseq.R +++ b/main/como/rscripts/rnaseq.R @@ -1,125 +1,109 @@ -suppressPackageStartupMessages(library("tidyverse")) -suppressPackageStartupMessages(library("limma")) +suppressPackageStartupMessages(library("biomaRt")) +suppressPackageStartupMessages(library("dplyr")) suppressPackageStartupMessages(library("edgeR")) suppressPackageStartupMessages(library("genefilter")) -suppressPackageStartupMessages(library("biomaRt")) +suppressPackageStartupMessages(library("limma")) +suppressPackageStartupMessages(library("readr")) +suppressPackageStartupMessages(library("readxl")) suppressPackageStartupMessages(library("sjmisc")) -suppressPackageStartupMessages(library("zFPKM")) suppressPackageStartupMessages(library("stringr")) -suppressPackageStartupMessages(library("readxl")) -suppressPackageStartupMessages(library("dplyr")) +suppressPackageStartupMessages(library("tidyverse")) +suppressPackageStartupMessages(library("zFPKM")) # Check if rlogs directory exists, From: https://stackoverflow.com/a/46008094 # Then prevent messy messages from repeatedly writing to juypter work_dir <- getwd() -r_log_directory <- str_interp("${work_dir}/logs") +r_log_directory <- stringr::str_interp("${work_dir}/logs") if (!dir.exists(r_log_directory)) { dir.create(r_log_directory) } zz <- file(file.path(r_log_directory, "rnaseq.Rout"), open = "wt") sink(zz, type = "message") +# Create a variable x set equal to 1 + + read_counts_matrix <- function(counts_matrix_filepath, config_filepath, info_filepath, context_name) { - config_object <- read_excel(config_filepath, sheet = context_name) # read configuration sheet - counts_matrix <- read.csv(counts_matrix_filepath, header = TRUE) %>% arrange(., genes) # read counts matrix - - gene_info <- read.csv(info_filepath) %>% - mutate(size = end_position - start_position) %>% - arrange(., ensembl_gene_id) %>% - filter(.$ensembl_gene_id %in% counts_matrix$genes) - - counts_matrix <- counts_matrix[(gene_info$entrezgene_id != "-"),] # remove unnamed genes - gene_info <- gene_info[(gene_info$entrezgene_id != "-"),] # remove unnamed genes - genes <- gene_info$entrezgene_id # get gene names - + config_object <- readxl::read_excel(config_filepath, sheet = context_name) + counts_matrix <- data.frame(readr::read_csv(file = counts_matrix_filepath)) %>% dplyr::arrange(., ensembl_gene_id) + gene_info <- data.frame(readr::read_csv(info_filepath)) %>% + dplyr::mutate(size = end_position - start_position) %>% # Calculate gene size + dplyr::arrange(.$ensembl_gene_id) %>% + dplyr::filter(.$entrez_gene_id != "-") %>% # Remove un-named genes + dplyr::filter(.$ensembl_gene_id %in% counts_matrix$ensembl_gene_id) # Only keep genes that are in counts_matrix + + # Select rows where "genes" in counts_matrix are also present in gene_info + counts_matrix <- counts_matrix[(counts_matrix$ensembl_gene_id %in% gene_info$ensembl_gene_id),] + entrez_genes <- gene_info$entrez_gene_id + # remove version numbers from ensembl id - for (i in seq_along(genes)) { - row <- genes[i] + for (i in seq_along(entrez_genes)) { + row <- entrez_genes[i] if (grepl("\\\\.", row)) { - gen <- unlist(str_split(row, "\\\\."))[1] - genes[i] <- gen + gen <- unlist(stringr::str_split(row, "\\\\."))[1] + entrez_genes[i] <- gen } } - + + + # initialize groups and pre-allocate space sample_metrics <- list() - groups <- unique(config_object$Group) - - # initialize groups + num_samples <- length(config_object$sample_name) + groups <- unique(config_object$study) for (group in groups) { - sample_metrics[[group]][["CountMatrix"]] <- genes - sample_metrics[[group]][["FragmentLengths"]] <- c() - sample_metrics[[group]][["SampleNames"]] <- c() - sample_metrics[[group]][["Layout"]] <- c() + sample_metrics[[group]] <- list( + CountMatrix = matrix(data = NA, nrow = nrow(counts_matrix), ncol = num_samples), + FragmentLengths = numeric(length = num_samples), + SampleNames = character(length = num_samples), + Layout = character(length = num_samples) + ) } - + # add to group count matrices and insert lists - for (i in seq_along(config_object$SampleName)) { - entry <- config_object$SampleName[i] # CELL-TYPE_S##R## - group <- config_object$Group[i] - - # These values will be added to the SampMetrix object - matrix_value <- counts_matrix[, entry] - fragment_length_value <- config_object$FragmentLength[i] - layout_value <- config_object$Layout[i] - sample_name_value <- entry - - - # In case insert_size is NULL, set it to 0 - if (is.na(fragment_length_value)) { - fragment_length_value <- 0 - } - - if (entry %in% colnames(counts_matrix)) { - - # get init values - sample_matrix <- sample_metrics[[group]][["CountMatrix"]] - fragment_lengths <- sample_metrics[[group]][["FragmentLengths"]] - sample_names <- sample_metrics[[group]][["SampleNames"]] - layouts <- sample_metrics[[group]][["Layout"]] - - # add replicate to values - sample_matrix <- cbind(sample_matrix, matrix_value) - fragment_lengths <- c(fragment_lengths, fragment_length_value) - layouts <- c(layouts, layout_value) - sample_names <- c(sample_names, sample_name_value) - - # update values - sample_metrics[[group]][["CountMatrix"]] <- sample_matrix - sample_metrics[[group]][["FragmentLengths"]] <- fragment_lengths - sample_metrics[[group]][["SampleNames"]] <- sample_names - sample_metrics[[group]][["Layout"]] <- layouts - - - } else { - print(paste(c(entry, " not found in count matrix."), collapse = "")) # inform that a sample is missing + for (i in seq_along(config_object$sample_name)) { + entry <- config_object$sample_name[i] # CELL-TYPE_S##R## + group <- config_object$study[i] + + # Test if `entry` is not in counts_matrix columns + if (!(entry %in% colnames(counts_matrix))) { + print(paste(entry, "not found in count matrix.")) + next } + + # In case insert_size is NULL, set it to 0 + fragment_length_value <- ifelse(is.na(config_object$fragment_length[i]), 0, config_object$fragment_length[i]) + + # update sample_metrics with current replicate values + col_index <- which(colnames(counts_matrix) == entry) # Use column index for faster access + sample_metrics[[group]][["CountMatrix"]][, i] <- as.numeric(counts_matrix[, col_index]) + sample_metrics[[group]][["FragmentLengths"]][i] <- fragment_length_value + sample_metrics[[group]][["SampleNames"]][i] <- entry + sample_metrics[[group]][["Layout"]][i] <- config_object$layout[i] } - + for (group in groups) { # for each study/batch group samp_mat <- sample_metrics[[group]][["CountMatrix"]] - samp_mat <- samp_mat[, -1] - samp_mat <- sapply(data.frame(samp_mat), as.numeric) - samp_mat <- as.matrix(samp_mat) # convert df to matrix - # samp_mat <- t(samp_mat) - + samp_mat <- apply(samp_mat, FUN = as.numeric, MARGIN = 2, simplify = FALSE) # '2' means apply over columns + write.csv(samp_mat, file = "/home/joshl/projects/COMO/main/samp_mat.csv") + samp_mat <- t(samp_mat) + colnames(samp_mat) <- sample_metrics[[group]][["SampleNames"]] # set column names to sample names sample_metrics[[group]][["CountMatrix"]] <- samp_mat # update counts matrix sample_metrics[[group]][["NumSamples"]] <- ncol(samp_mat) # set number of samples - sample_metrics[[group]][["Entrez"]] <- as.character(gene_info$entrezgene_id) # store entrez ids + sample_metrics[[group]][["Entrez"]] <- as.character(gene_info$entrez_gene_id) # store entrez ids sample_metrics[[group]][["GeneSizes"]] <- gene_info$size # store gene size sample_metrics[[group]][["StudyNumber"]] <- group } - return(sample_metrics) } calculate_tpm <- function(sample_metrics) { - - for (i in 1:length(sample_metrics)) { + + for (i in seq_along(sample_metrics)) { count_matrix <- sample_metrics[[i]][["CountMatrix"]] gene_size <- sample_metrics[[i]][["GeneSizes"]] - tpm_matrix <- do.call(cbind, lapply(1:ncol(count_matrix), function(j) { - rate = log(count_matrix[, j]) - log(gene_size[j]) - denom = log(sum(exp(rate))) + tpm_matrix <- do.call(cbind, lapply(seq_len(ncol(count_matrix)), function(j) { + rate <- log(count_matrix[, j]) - log(gene_size[j]) + denom <- log(sum(exp(rate))) exp(rate - denom + log(1e6)) })) colnames(tpm_matrix) <- colnames(count_matrix) @@ -132,47 +116,52 @@ calculate_tpm <- function(sample_metrics) { calculate_fpkm <- function(sample_metrics) { - for (i in 1:length(sample_metrics)) { + for (i in seq_along(sample_metrics)) { layout <- sample_metrics[[i]][["Layout"]] # get layout - - # normalize with fpkm if paired, rpkm if single end, kill if something else is specified - stopifnot(layout[1] == "paired-end" || layout[1] == "single-end") + if (layout[1] == "paired-end") { # fpkm + # Print components of sample_metrics[[i]], which is of type list count_matrix <- sample_metrics[[i]][["CountMatrix"]] gene_size <- sample_metrics[[i]][["GeneSizes"]] mean_fragment_lengths <- sample_metrics[[i]][["FragmentLengths"]] - fpkm_matrix <- do.call(cbind, lapply(1:ncol(count_matrix), function(j) { - eff_len = gene_size - mean_fragment_lengths[j] + 1 # plus one to prevent div by 0 - N = sum(count_matrix[, j]) - exp(log(count_matrix[, j]) + log(1e9) - - log(eff_len) - - log(N)) - })) + + fpkm_matrix <- matrix(nrow = nrow(count_matrix), ncol = ncol(count_matrix)) + print(class(count_matrix)) + print(head(colSums(count_matrix))) + for (j in seq_len(ncol(count_matrix))) { + eff_len <- gene_size - mean_fragment_lengths[j] + 1 # plus one to prevent div by 0 + + print(paste("j:", j)) + print(paste("ncol:", ncol(count_matrix))) + N <- sum(count_matrix[, j]) + + fpkm_matrix[, j] <- exp(log(count_matrix[, j]) + log(1e9) - log(eff_len) - log(N)) + } fpkm_matrix[is.nan(fpkm_matrix)] <- 0 colnames(fpkm_matrix) <- colnames(count_matrix) sample_metrics[[i]][["FPKM_Matrix"]] <- fpkm_matrix - - } else { # rpkm + + } else if (layout[1] == "single-end" ) { # rpkm count_matrix <- sample_metrics[[i]][["CountMatrix"]] gene_size <- sample_metrics[[i]][["GeneSizes"]] - rpkm_matrix <- do.call(cbind, lapply(1:ncol(count_matrix), function(j) { - rate = log(count_matrix[, j]) - log(gene_size[j]) + rpkm_matrix <- do.call(cbind, lapply(seq_len(ncol(count_matrix)), function(j) { + rate <- log(count_matrix[, j]) - log(gene_size[j]) exp(rate - log(sum(count_matrix[, j])) + log(1e9)) })) rpkm_matrix[is.nan(rpkm_matrix)] <- 0 colnames(rpkm_matrix) <- colnames(count_matrix) sample_metrics[[i]][["FPKM_Matrix"]] <- rpkm_matrix - + } else { + stop(paste0("Invalid layout for sample '", sample_metrics[[i]][["StudyNumber"]], "'. Must be paired-end or single-end")) } } - return(sample_metrics) } calculate_z_score <- function(sample_metrics, norm_tech) { - for (i in 1:length(sample_metrics)) { + for (i in seq_along(sample_metrics)) { if (norm_tech == "CPM") { tmat <- sample_metrics[[i]][["CPM_Matrix"]] } else if (norm_tech == "TPM") { @@ -180,7 +169,7 @@ calculate_z_score <- function(sample_metrics, norm_tech) { } zmat <- matrix(nrow = nrow(tmat), ncol = ncol(tmat)) - for (j in 1:ncol(tmat)) { + for (j in seq_len(ncol(tmat))) { tvec <- tmat[, j] logvec <- log2(tvec) logvec[is.infinite(logvec)] <- NA @@ -201,14 +190,14 @@ cpm_filter <- function(sample_metrics, filt_options, context_name, prep) { N_exp <- filt_options$replicate_ratio N_top <- filt_options$replicate_ratio_high min.count <- filt_options$min_count - for (i in 1:length(sample_metrics)) { + for (i in seq_along(sample_metrics)) { study_number <- sample_metrics[[i]][["StudyNumber"]] counts <- sample_metrics[[i]][["CountMatrix"]] ent <- sample_metrics[[i]][["Entrez"]] size <- sample_metrics[[i]][["GeneSizes"]] lib.size <- colSums(counts) - CPM <- cpm(counts, lib.size = lib.size) + CPM <- edgeR::cpm(counts, lib.size = lib.size) cpm_fname <- file.path(work_dir, "data", "results", context_name, prep, paste0("CPM_Matrix_", prep, "_", study_number, ".csv")) write_cpm <- cbind(ent, CPM) write.csv(write_cpm, cpm_fname, row.names = FALSE) @@ -217,7 +206,7 @@ cpm_filter <- function(sample_metrics, filt_options, context_name, prep) { top.samples <- round(N_top * ncol(counts)) test_bools <- data.frame(gene = ent) - for (j in 1:ncol(CPM)) { + for (j in seq_len(ncol(CPM))) { cutoff <- ifelse(min.count == "default", 10e6 / (median(sum(counts[, j]))), 1e6 * min.count / (median(sum(counts[, j])))) @@ -253,7 +242,7 @@ TPM_quant_filter <- function(sample_metrics, filt_options, context_name, prep) { quant <- filt_options$quantile sample_metrics <- calculate_tpm(sample_metrics) - for (i in 1:length(sample_metrics)) { + for (i in seq_along(sample_metrics)) { study_number <- sample_metrics[[i]][["StudyNumber"]] counts <- sample_metrics[[i]][["CountMatrix"]] @@ -268,7 +257,7 @@ TPM_quant_filter <- function(sample_metrics, filt_options, context_name, prep) { top.samples <- round(N_top * ncol(tpm)) test_bools <- data.frame(gene = ent) - for (j in 1:ncol(tpm)) { + for (j in seq_len(ncol(tpm))) { tpm_q <- tpm[, j] tpm_q <- tpm_q[tpm_q > 0] q_cutoff <- quantile(tpm_q, prob = 1 - quant / 100) @@ -304,11 +293,11 @@ zfpkm_filter <- function(sample_metrics, filt_options, context_name, prep) { N_exp <- filt_options$replicate_ratio # ratio replicates for active N_top <- filt_options$replicate_ratio_high # ratio of replicates for high-confidence cutoff <- filt_options$min_zfpkm - + sample_metrics <- calculate_fpkm(sample_metrics) for (i in seq_along(sample_metrics)) { study_number <- sample_metrics[[i]][["StudyNumber"]] - + entrez_ids <- sample_metrics[[i]][["Entrez"]] # get entrez ids fpkm_matrix <- sample_metrics[[i]][["FPKM_Matrix"]] # get fpkm matrix fpkm_df <- data.frame(fpkm_matrix) # convert to df @@ -317,16 +306,16 @@ zfpkm_filter <- function(sample_metrics, filt_options, context_name, prep) { write_fpkm <- cbind(entrez_ids, fpkm_df) colnames(write_fpkm)[1] <- "ENTREZ_GENE_ID" write.csv(write_fpkm, fpkm_filename, row.names = FALSE) - + minimums <- fpkm_df == 0 - nas <- is.na(fpkm_df) == 1 - + na_values <- is.na(fpkm_df) == 1 + # calculate zFPKM - zmat <- zFPKM(fpkm_df, min_thresh = 0, assayName = "FPKM") + zmat <- zFPKM::zFPKM(fpkm_df, min_thresh = 0, assayName = "FPKM") zmat[minimums] <- -4 # instead of -inf set to lower limit zfpkm_fname <- file.path(work_dir, "data", "results", context_name, prep, paste0("zFPKM_Matrix_", prep, "_", study_number, ".csv")) - write_zfpkm <- cbind(entrez_ids, zmat) + write_zfpkm <- dplyr::bind_cols(entrez_ids, zmat) colnames(write_zfpkm)[1] <- "ENTREZ_GENE_ID" write.csv(write_zfpkm, zfpkm_fname, row.names = FALSE) @@ -338,7 +327,7 @@ zfpkm_filter <- function(sample_metrics, filt_options, context_name, prep) { zfpkm_plotname <- file.path(zfpkm_plot_dir, paste0("zFPKM_plot_", study_number, ".pdf")) pdf(zfpkm_plotname) - zFPKMPlot(fpkm_df, min_thresh = min(fpkm_df), assayName = "FPKM") + zFPKM::zFPKMPlot(fpkm_df, min_thresh = min(fpkm_df), assayName = "FPKM") dev.off() min.samples <- round(N_exp * ncol(zmat)) # min number of samples for active @@ -356,8 +345,7 @@ zfpkm_filter <- function(sample_metrics, filt_options, context_name, prep) { keep_top <- genefilter::genefilter(zmat, flist_top) sample_metrics[[i]][["Entrez_hc"]] <- entrez_ids[keep_top] } - - + return(sample_metrics) } @@ -367,43 +355,46 @@ umi_filter <- function(sample_metrics, filt_options, context_name) { N_exp <- filt_options$replicate_ratio # ratio replicates for active N_top <- filt_options$replicate_ratio_high # ratio of replicates for high-confidence cutoff <- filt_options$min_zfpkm - + + #sample_metrics <- calculate_fpkm(sample_metrics) - for (i in 1:length(sample_metrics)) { + for (i in seq_along(sample_metrics)) { study_number <- sample_metrics[[i]][["StudyNumber"]] - ent <- sample_metrics[[i]][["Entrez"]] # get entrez ids - umat <- sample_metrics[[i]][["CountMatrix"]] # get fpkm matrix - udf <- data.frame(umat) # convert to df - udf[rowSums(udf[]) > 0,] - minimums <- udf == 0 - nas <- is.na(udf) == 1 - zmat <- zFPKM(udf, min_thresh = 0, assayName = "UMI") # calculate zFPKM + + entrez_ids <- sample_metrics[[i]][["Entrez"]] # get entrez ids + count_matrix <- data.frame(sample_metrics[[i]][["CountMatrix"]]) + # Convert count matrix to numeric + count_matrix <- sapply(count_matrix, as.numeric) + count_matrix[rowSums(count_matrix[]) > 0,] + minimums <- count_matrix == 0 + + zmat <- zFPKM::zFPKM(count_matrix, min_thresh = 0, assayName = "UMI") zmat[minimums] <- -4 # instead of -inf set to lower limit - zumi_fname <- file.path(work_dir, "data", "results", context_name, prep, paste0("zUMI_Matrix_", prep, "_", study_number, ".csv")) - # zumi_fname <- file.path("/home", username, "main", "data", "results", context_name, prep, paste0("zUMI_Matrix_", prep, "_", study_number, ".csv")) - - write_zumi <- cbind(ent, zmat) + + write_zumi <- dplyr::bind_cols(entrez_ids, zmat) + # write_zumi <- cbind(ent, zmat) colnames(write_zumi)[1] <- "ENTREZ_GENE_ID" + zumi_fname <- file.path(work_dir, "data", "results", context_name, prep, paste0("zUMI_Matrix_", prep, "_", study_number, ".csv")) write.csv(write_zumi, zumi_fname, row.names = FALSE) - + zumi_plot_dir <- file.path(work_dir, "data", "results", context_name, prep, "figures") # zumi_plot_dir <- file.path("/home", username, "main", "data", "results", context_name, prep, "figures") - + if (!file.exists(zumi_plot_dir)) { dir.create(zumi_plot_dir) } - + batch_size <- 12 - plot_batches <- ceiling(ncol(udf) / batch_size) - + plot_batches <- ceiling(ncol(count_matrix) / batch_size) + if (plot_batches < 2) { zumi_plotname <- file.path(zumi_plot_dir, paste0("zumi_plot_", study_number, ".pdf")) pdf(zumi_plotname) - zFPKMPlot(udf, min_thresh = min(udf), assayName = "UMI") + zFPKM::zFPKMPlot(count_matrix, min_thresh = min(count_matrix), assayName = "UMI") dev.off() - + } else { - + for (j in 1:(plot_batches - 1)) { zumi_plotname <- file.path(zumi_plot_dir, paste0("zumi_plot_", study_number, "_", j, ".pdf")) pdf(zumi_plotname) @@ -411,38 +402,38 @@ umi_filter <- function(sample_metrics, filt_options, context_name) { jmin <- (batch_size * (j - 1)) + 1 jmax <- batch_size * j samps <- jmin:jmax - - while (samps[length(samps)] > ncol(udf)) { - samps <- samps[1:length(samps) - 1] + + while (samps[length(samps)] > ncol(count_matrix)) { + samps <- samps[seq_along(samps) - 1] } - - zFPKMPlot( - udf[, samps], + + zFPKM::zFPKMPlot( + count_matrix[, samps], min_thresh = 0, assayName = "UMI" ) dev.off() } } - + min.samples <- round(N_exp * ncol(zmat)) # min number of samples for active top.samples <- round(N_top * ncol(zmat)) # top number of samples for high-confidence - + # active genes f1 <- genefilter::kOverA(min.samples, cutoff) flist <- genefilter::filterfun(f1) keep <- genefilter::genefilter(zmat, flist) - sample_metrics[[i]][["Entrez"]] <- ent[keep] - + sample_metrics[[i]][["Entrez"]] <- entrez_ids[keep] + # top percentile genes f1_top <- genefilter::kOverA(top.samples, cutoff) flist_top <- genefilter::filterfun(f1_top) keep_top <- genefilter::genefilter(zmat, flist_top) - - sample_metrics[[i]][["Entrez_hc"]] <- ent[keep_top] - + + sample_metrics[[i]][["Entrez_hc"]] <- entrez_ids[keep_top] + } - + return(sample_metrics) } @@ -458,88 +449,52 @@ filter_counts <- function(sample_metrics, technique, filt_options, context_name, } save_rnaseq_tests <- function( - counts_matrix_file, - config_filepath, - out_file, - info_file, - context_name, - prep = "total", - replicate_ratio = 0.5, - batch_ratio = 0.5, - replicate_ratio_high = 0.9, - batch_ratio_high = 0.9, - technique = "quantile", - quantile = 0.9, - min_count = 10, - min_zfpkm = -3 + counts_matrix_file, + config_filepath, + out_file, + info_file, + context_name, + prep = "total", + replicate_ratio = 0.5, + batch_ratio = 0.5, + replicate_ratio_high = 0.9, + batch_ratio_high = 0.9, + technique = "quantile", + quantile = 0.9, + min_count = 10, + min_zfpkm = -3 ) { # condense filter options filt_options <- list() - if (exists("replicate_ratio")) { - filt_options$replicate_ratio <- replicate_ratio - } else { - filt_options$replicate_ratio <- 0.2 - } - if (exists("batch_ratio")) { - filt_options$batch_ratio <- batch_ratio - } else { - filt_options$batch_ratio <- 0.5 - } - if (exists("quantile")) { - filt_options$quantile <- quantile - } else { - filt_options$quantile <- 50 - } - if (exists("min_count")) { - filt_options$min_count <- min_count - } else { - filt_options$min_count <- 1 - } - if (exists("min_zfpkm")) { - filt_options$min_zfpkm <- min_zfpkm - } else { - filt_options$min_zfpkm <- -3 - } - if (exists("replicate_ratio_high")) { - filt_options$replicate_ratio_high <- replicate_ratio_high - } else { - filt_options$replicate_ratio_high <- 0.9 - } - if (exists("batch_ratio_high")) { - filt_options$batch_ratio_high <- batch_ratio_high - } else { - filt_options$batch_ratio_high <- 0.9 - } + filt_options$replicate_ratio <- replicate_ratio + filt_options$batch_ratio <- batch_ratio + filt_options$quantile <- quantile + filt_options$min_count <- min_count + filt_options$min_zfpkm <- min_zfpkm + filt_options$replicate_ratio_high <- replicate_ratio_high + filt_options$batch_ratio_high <- batch_ratio_high if (prep == "scrna") { technique <- "umi" print("Note: Single cell filtration does not normalize and assumes counts are counted with UMI") } - - sample_metrics <- read_counts_matrix(counts_matrix_file, config_filepath, info_file, context_name) # read count matrix - entrez_all <- sample_metrics[[1]][["Entrez"]] #get entrez ids - - sample_metrics <- filter_counts(sample_metrics, technique, filt_options, context_name, prep) # normalize and filter count - expressedGenes <- c() - topGenes <- c() - for (i in 1:length(sample_metrics)) { # get high confidence and expressed genes for each study/batch number - expressedGenes <- c(expressedGenes, sample_metrics[[i]][["Entrez"]]) - topGenes <- c(topGenes, sample_metrics[[i]][["Entrez_hc"]]) - } + + print("1") sample_metrics <- read_counts_matrix(counts_matrix_file, config_filepath, info_file, context_name) # read count matrix - + print("2") entrez_all <- sample_metrics[[1]][["Entrez"]] #get entrez ids sample_metrics <- filter_counts(sample_metrics, technique, filt_options, context_name, prep) # normalize and filter count + print("3") expressedGenes <- c() topGenes <- c() - - for (i in 1:length(sample_metrics)) { # get high confidence and expressed genes for each study/batch number + for (i in seq_along(sample_metrics)) { # get high confidence and expressed genes for each study/batch number expressedGenes <- c(expressedGenes, sample_metrics[[i]][["Entrez"]]) topGenes <- c(topGenes, sample_metrics[[i]][["Entrez_hc"]]) } - + print("4") + expMat <- as.data.frame(table(expressedGenes)) # convert expression to df topMat <- as.data.frame(table(topGenes)) # convert high confidence to df nc <- length(sample_metrics) # number of columns @@ -547,14 +502,16 @@ save_rnaseq_tests <- function( topMat <- cbind(topMat, "Prop" = topMat$Freq / nc) # calculate proportion of studies/batch high-confidence sample_metrics[["ExpressionMatrix"]] <- expMat # store expression matrix for saving sample_metrics[["TopMatrix"]] <- topMat # store high confidence matrix for saving + print("8") # get genes which are expressed and high-confidence according to use defined ratio sample_metrics[["ExpressedGenes"]] <- as.character(expMat$expressedGenes[expMat$Prop >= batch_ratio]) sample_metrics[["TopGenes"]] <- as.character(topMat$topGenes[topMat$Prop >= batch_ratio_high]) - + print("9") + # create a table to write gene expression and high confidence to write_table <- data.frame(entrez_all) write_table <- cbind(write_table, rep(0, nrow(write_table))) - for (i in 1:nrow(write_table)) { + for (i in seq_len(nrow(write_table))) { if (as.character(write_table[i, 1]) %in% as.character(sample_metrics$ExpressedGenes)) { write_table[i, 2] <- 1 } @@ -562,8 +519,20 @@ save_rnaseq_tests <- function( write_table[i, 3] <- 1 } } + print("10") header <- c("ENTREZ_GENE_ID", "expressed", "high") #write_table <- rbind(header, write_table) colnames(write_table) <- header write.csv(write_table, out_file, row.names = FALSE, col.names = FALSE) + print("11") + print(head(write_table)) } + +save_rnaseq_tests( + counts_matrix_file = "/home/joshl/projects/COMO/main/data/data_matrices/naiveB/gene_counts_matrix_total_naiveB.csv", + config_filepath = "/home/joshl/projects/COMO/main/data/config_sheets/trnaseq_data_inputs_auto.xlsx", + out_file = "/home/joshl/projects/COMO/rnaseq_r_out.csv", + info_file = "/home/joshl/projects/COMO/main/data/gene_info.csv", + context_name = "naiveB", + technique = "zfpkm" +) \ No newline at end of file diff --git a/main/como/utils.py b/main/como/utils.py new file mode 100644 index 00000000..9668a34d --- /dev/null +++ b/main/como/utils.py @@ -0,0 +1,260 @@ +from __future__ import annotations + +import contextlib +import io +import sys +from collections.abc import Iterator +from enum import Enum +from pathlib import Path +from typing import ClassVar + +import aiofiles +import pandas as pd +from fast_bioservices import BioDBNet, Output, Taxon +from fast_bioservices.pipeline import ( + determine_gene_type, + ensembl_to_gene_id_and_symbol, + gene_id_to_ensembl_and_gene_symbol, + gene_symbol_to_ensembl_and_gene_id, +) +from loguru import logger + +__all__ = ["Compartments", "split_gene_expression_data", "stringlist_to_list", "suppress_stdout"] + + +class Algorithm(Enum): + GIMME = "GIMME" + FASTCORE = "FASTCORE" + IMAT = "IMAT" + TINIT = "TINIT" + + @staticmethod + def from_string(value: str) -> Algorithm: + match value.lower(): + case "gimme": + return Algorithm.GIMME + case "fastcore": + return Algorithm.FASTCORE + case "imat": + return Algorithm.IMAT + case "tinit": + return Algorithm.TINIT + case _: + raise ValueError(f"Unknown solver: {value}") + + +class Compartments: + """Convert from compartment "long-hand" to "short-hand". + + Shorthand from: https://cobrapy.readthedocs.io/en/latest/_modules/cobra/medium/annotations.html + + "Extracellular" -> "e" + "golgi" -> "g" + """ + + SHORTHAND: ClassVar[dict[str, list[str]]] = { + "ce": ["cell envelope"], + "c": [ + "cytoplasm", + "cytosol", + "default", + "in", + "intra cellular", + "intracellular", + "intracellular region", + "intracellular space", + ], + "er": ["endoplasmic reticulum"], + "erm": ["endoplasmic reticulum membrane"], + "e": [ + "extracellular", + "extraorganism", + "out", + "extracellular space", + "extra organism", + "extra cellular", + "extra-organism", + "external", + "external medium", + ], + "f": ["flagellum", "bacterial-type flagellum"], + "g": ["golgi", "golgi apparatus"], + "gm": ["golgi membrane"], + "h": ["chloroplast"], + "l": ["lysosome"], + "im": ["mitochondrial intermembrane space"], + "mm": ["mitochondrial membrane"], + "m": ["mitochondrion", "mitochondria"], + "n": ["nucleus"], + "p": ["periplasm", "periplasmic space"], + "x": ["peroxisome", "glyoxysome"], + "u": ["thylakoid"], + "vm": ["vacuolar membrane"], + "v": ["vacuole"], + "w": ["cell wall"], + "s": ["eyespot", "eyespot apparatus", "stigma"], + } + + _REVERSE_LOOKUP: ClassVar[dict[str, list[str]]] = { + value.lower(): key for key, values in SHORTHAND.items() for value in values + } + + @classmethod + def get(cls, longhand: str) -> str | None: + """Get the short-hand compartment name from the long-hand name.""" + return cls._REVERSE_LOOKUP.get(longhand.lower(), None) + + +def stringlist_to_list(stringlist: str | list[str]) -> list[str]: + """Convert a string from the command line into a Python list. + + In doing so, we must deprecate the use of the current method + + If '[' and ']' are present in the first and last items of the list, + assume we are using the "old" method of providing context names + + :param stringlist: The "string list" gathered from the command line. Example input: "['mat', 'xml', 'json']" + """ + if isinstance(stringlist, str): + if stringlist.startswith("[") and stringlist.endswith("]"): + # Remove any brackets from the first and last items; replace quotation marks and commas with nothing + new_list: list[str] = stringlist.strip("[]").replace("'", "").replace(" ", "").split(",") + + # Show a warning if more than one item is present in the list (this means we are using the old method) + logger.critical( + "DeprecationWarning: Please use the new method of providing context names, " + "i.e. --output-filetypes 'type1 type2 type3'." + ) + logger.critical( + "If you are using COMO, this can be done by setting the 'context_names' variable to a " + "simple string separated by spaces. Here are a few examples!" + ) + logger.critical("context_names = 'cellType1 cellType2 cellType3'") + logger.critical("output_filetypes = 'output1 output2 output3'") + logger.critical( + "\nYour current method of passing context names will be removed in the future. " + "Update your variables above accordingly!\n\n" + ) + + else: + new_list: list[str] = stringlist.split(" ") + + return new_list + return stringlist + + +def split_gene_expression_data(expression_data: pd.DataFrame, recon_algorithm: Algorithm | None = None): + """Split the gene expression data into single-gene and multiple-gene names. + + :param expression_data: The gene expression data to map + :param recon_algorithm: The recon algorithm used to generate the gene expression data + :return: + """ + expression_data.columns = [c.lower() for c in expression_data.columns] + if recon_algorithm in {Algorithm.IMAT, Algorithm.TINIT}: + expression_data.rename(columns={"combine_z": "active"}, inplace=True) + + expression_data = expression_data[["entrez_gene_id", "active"]] + expression_data.loc[:, "entrez_gene_id"] = expression_data["entrez_gene_id"].astype(str) + single_gene_names = expression_data[~expression_data["entrez_gene_id"].str.contains("//")] + multiple_gene_names = expression_data[expression_data["entrez_gene_id"].str.contains("//")] + split_gene_names = multiple_gene_names.assign( + entrez_gene_id=multiple_gene_names["entrez_gene_id"].str.split("///") + ).explode("entrez_gene_id") + + gene_expressions = pd.concat([single_gene_names, split_gene_names], axis=0, ignore_index=True) + gene_expressions.set_index("entrez_gene_id", inplace=True) + return gene_expressions + + +@contextlib.contextmanager +def suppress_stdout() -> Iterator[None]: + """Suppress stdout output from the current context. + + :return: The context manager + """ + with io.StringIO() as buffer: + try: + sys.stdout = buffer + yield + finally: + sys.stdout = sys.__stdout__ + + +async def _format_determination( + biodbnet: BioDBNet, *, requested_output: Output | list[Output], input_values: list[str], taxon: Taxon +) -> pd.DataFrame: + """Determine the data type of the given input values (i.e., Entrez Gene ID, Gene Symbol, etc.). + + :param biodbnet: The BioDBNet to use for deter + :param requested_output: The data type to generate (of type `Output`) + :param input_values: The input values to determine + :param taxon: The Taxon ID + :return: A pandas DataFrame + """ + requested_output = [requested_output] if isinstance(requested_output, Output) else requested_output + cohersion = (await biodbnet.db_find(values=input_values, output_db=requested_output, taxon=taxon)).drop( + columns=["Input Type"] + ) + cohersion.columns = pd.Index(["input_value", *[o.value.replace(" ", "_").lower() for o in requested_output]]) + return cohersion + + +async def _async_read_csv(path: Path, **kwargs) -> pd.DataFrame: + """Asynchronously reads a CSV file and returns a pandas DataFrame. + + :param path: The path to read from + :param kwargs: Additional arguments to pass to pandas.read_csv + :return: A pandas DataFrame + """ + if not path.exists(): + raise FileNotFoundError(f"File {path} does not exist") + if path.suffix not in {".csv", ".tsv"}: + raise ValueError(f"File {path} is not a CSV file") + + kwargs.setdefault("sep", "," if path.suffix == ".csv" else "\t") + async with aiofiles.open(path) as f: + content = await f.read() + return pd.read_csv(io.StringIO(content), **kwargs) + + +async def _async_read_excel(path: Path, **kwargs) -> pd.DataFrame: + """Asynchronously reads an Excel file and returns a pandas DataFrame. + + :param path: The path to read from + :param kwargs: Additional arguments to pass to pandas.read_excel + :return: A pandas DataFrame + """ + if not path.exists(): + raise FileNotFoundError(f"File {path} does not exist") + if path.suffix not in {".xls", ".xlsx"}: + raise ValueError(f"File {path} is not an Excel file") + + async with aiofiles.open(path, "rb") as f: + content = await f.read() + return pd.read_excel(io.StringIO(content.decode()), **kwargs) + + +def is_notebook() -> bool: + """Check if the current environment is a Jupyter Notebook. + + :returns: True if the current environment is a Jupyter Notebook, False otherwise. + """ + try: + from IPython import get_ipython + + return get_ipython() is not None + except ModuleNotFoundError: + return False + + +async def convert_gene_data(values: list[str], taxon_id: int | str | Taxon) -> pd.DataFrame: + gene_type = await determine_gene_type(values) + if all(v == "gene_symbol" for v in gene_type.values()): + return await gene_symbol_to_ensembl_and_gene_id(values, taxon=taxon_id) + elif all(v == "ensembl_gene_id" for v in gene_type.values()): + return await ensembl_to_gene_id_and_symbol(ids=values, taxon=taxon_id) + elif all(v == "entrez_gene_id" for v in gene_type.values()): + return await gene_id_to_ensembl_and_gene_symbol(ids=values, taxon=taxon_id) + else: + raise ValueError("Gene data must be of the same type (i.e., all Ensembl, Entrez, or Gene Symbols)") diff --git a/main/data/boundary_rxns/naiveB_boundary_rxns.csv b/main/data/boundary_rxns/naiveB_boundary_rxns.csv index 9f0f9385..7130c9be 100644 --- a/main/data/boundary_rxns/naiveB_boundary_rxns.csv +++ b/main/data/boundary_rxns/naiveB_boundary_rxns.csv @@ -67,4 +67,4 @@ Exchange,orn,Extracellular,-1,1000 Exchange,orn_D,Extracellular,-1,1000 Exchange,so4,Extracellular,-1000,1000 Exchange,ribflv,Extracellular,-1,1000 -Exchange,Lcystin,Extracellular,-1,1000 +Exchange,Lcystin,Extracellular,-1,1000 \ No newline at end of file diff --git a/main/data/config_sheets/scrnaseq_data_inputs_template.xlsx b/main/data/config_sheets/scrnaseq_data_inputs_template.xlsx deleted file mode 100644 index f811af95..00000000 Binary files a/main/data/config_sheets/scrnaseq_data_inputs_template.xlsx and /dev/null differ diff --git a/main/data/data_matrices/naiveB/gene_counts_matrix_full_naiveB.csv b/main/data/data_matrices/naiveB/gene_counts_matrix_full_naiveB.csv index 8bb68e30..a5d14856 100644 --- a/main/data/data_matrices/naiveB/gene_counts_matrix_full_naiveB.csv +++ b/main/data/data_matrices/naiveB/gene_counts_matrix_full_naiveB.csv @@ -1,61542 +1,61542 @@ 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+ENSG00000210195,6,19,3 +ENSG00000210196,121,210,565 +ENSG00000274175,0,0,0 +ENSG00000273554,0,0,0 +ENSG00000275869,0,0,0 +ENSG00000278510,0,0,0 +ENSG00000273843,0,0,0 +ENSG00000277836,6,11,1 +ENSG00000278673,5,12,8 +ENSG00000277890,0,0,0 +ENSG00000275249,0,0,0 +ENSG00000274792,0,0,0 +ENSG00000276872,0,0,0 +ENSG00000276760,0,0,0 +ENSG00000277927,0,0,0 +ENSG00000278457,0,0,0 +ENSG00000278294,0,0,0 +ENSG00000276711,0,0,0 +ENSG00000278704,0,0,0 +ENSG00000277400,0,0,0 +ENSG00000274847,0,0,0 +ENSG00000276932,0,0,0 +ENSG00000277428,0,0,0 +ENSG00000276256,0,1,1 +ENSG00000276312,0,0,0 +ENSG00000276700,0,0,0 +ENSG00000275757,0,0,0 +ENSG00000273739,0,0,0 +ENSG00000273748,18,23,17 +ENSG00000274202,0,0,0 +ENSG00000278625,0,0,0 +ENSG00000278573,0,0,0 +ENSG00000277196,0,0,0 +ENSG00000278817,0,0,0 +ENSG00000276017,0,0,0 +ENSG00000277630,0,0,0 +ENSG00000278793,0,0,0 +ENSG00000276197,0,0,0 +ENSG00000274917,0,0,0 +ENSG00000273730,0,0,0 +ENSG00000273937,0,0,0 +ENSG00000278384,5,2,11 +ENSG00000278633,0,0,0 +ENSG00000278066,0,0,0 +ENSG00000277374,0,0,0 +ENSG00000275661,0,0,0 +ENSG00000276351,0,0,0 +ENSG00000276345,0,0,0 +ENSG00000273532,0,0,0 +ENSG00000275063,95,89,1 +ENSG00000277856,40,68,4 +ENSG00000271254,15,54,4 +ENSG00000275987,0,0,0 +ENSG00000268674,0,0,0 +ENSG00000277475,0,4,0 +ENSG00000275405,0,0,0 diff --git a/main/data/data_matrices/placeholder/placeholder_empty_data.csv b/main/data/data_matrices/placeholder/placeholder_empty_data.csv deleted file mode 100644 index 486611ff..00000000 --- a/main/data/data_matrices/placeholder/placeholder_empty_data.csv +++ /dev/null @@ -1,2 +0,0 @@ -ENTREZ_GENE_ID,expressed,top -0,0,0 diff --git a/main/tests/environment.yaml b/main/tests/environment.yaml deleted file mode 100644 index 795a2383..00000000 --- a/main/tests/environment.yaml +++ /dev/null @@ -1,23 +0,0 @@ -name: como_tests -channels: - - conda-forge - - bioconda - - r - - defaults -dependencies: - - aioftp==0.21.3 - - conda-forge::bioservices~=1.11.2 - - flake8==5.0.4 - - pandas==1.4.4 - - python==3.10.10 - - pytest-asyncio==0.19.0 - - pytest-localftpserver==1.1.3 - - pytest-mock==3.8.2 - - pytest==7.1.3 - - rpy2==3.4.5 - - toml==0.10.2 - - tqdm==4.64.1 - - pip==22.1.2 - - pip: - - git+https://github.com/JoshLoecker/fast_bioservices - - GEOparse==2.0.3 diff --git a/main/tests/test_execution.py b/main/tests/test_execution.py deleted file mode 100644 index 30f782fd..00000000 --- a/main/tests/test_execution.py +++ /dev/null @@ -1,3 +0,0 @@ -def test_imports(): - from como.como_utilities import Compartments, stringlist_to_list, split_gene_expression_data - from como.project import Config diff --git a/pyproject.toml b/pyproject.toml index f8773cca..2c6d42c5 100644 --- a/pyproject.toml +++ b/pyproject.toml @@ -1,41 +1,52 @@ -[build-system] -requires = ["hatchling", "hatch-vcs"] -build-backend = "hatchling.build" - [project] name = "COMO" dynamic = ["version"] -#readme = "README.md" -requires-python = ">=3.8" +requires-python = ">=3.10,<3.13" dependencies = [ - "cobra", - "fast-bioservices>=0.3.4", - "cobamp @ git+https://github.com/JoshLoecker/cobamp.git", - "geoparse", - "numpy", - "openpyxl", - "pandas", - "rpy2", - "sqlalchemy", + "cobra>=0.28.0", + "fast-bioservices>=0.3.9", + "gurobipy>=11.0", + "kaleido==0.2.1", + "loguru>=0.7.2", + "pandas>=1.3.5", + "plotly>=5.24.1", + "scanpy>=1.9.8", + "scipy>=1.7.3", + "scikit-learn>=1.5.2", + "setuptools<60.0", + "openpyxl>=3.1.5", + "aiofiles>=24.1.0", + "aioftp>=0.23.1", "troppo", + "cobamp", ] -[tool.ruff] -line-length = 150 +[build-system] +requires = ["hatchling"] +build-backend = "hatchling.build" [tool.hatch.version] -source = "vcs" +path = "main/como/__init__.py" [tool.hatch.build] packages = ["main/como"] -[tool.hatch.build.hooks.vcs] -version-file = "_version.py" - [tool.hatch.metadata] -allow-direct-references=true +allow-direct-references = true [tool.pytest.ini_options] -pythonpath = [ - "main/src" +pythonpath = [ "main/src" ] + +[tool.uv] +dev-dependencies = [ + "commitlint>=1.3.0", + "pytest-asyncio>=0.24.0", + "pytest>=8.3.3", + "ruff>=0.8.0", + "hypothesis>=6.122.1", + "pytest-cov>=6.0.0", ] + +[tool.uv.sources] +troppo = { git = "https://github.com/JoshLoecker/troppo", rev = "update_dependencies" } +cobamp = { git = "https://github.com/JoshLoecker/cobamp", rev = "update_packages" } diff --git a/ruff.toml b/ruff.toml new file mode 100644 index 00000000..9591bda3 --- /dev/null +++ b/ruff.toml @@ -0,0 +1,50 @@ +line-length = 120 +extend-include = ["docs/**/*.py", "tests/**/*.py"] +exclude = ["__init__.py", "main/COMO.ipynb"] + +[format] +quote-style = "double" +docstring-code-format = true + +[lint] +# Linting rules: https://docs.astral.sh/ruff/rules/ +select = [ + "A", # do not use python builtins for variables or parameters; https://pypi.org/project/flake8-builtins/ + "ASYNC", # identify asynchronous-related problems; https://pypi.org/project/flake8-async/ + "B", # find likely bugs and design problems; https://pypi.org/project/flake8-bugbear/ + "C4", # create better list, set, & dict comprehensions; https://pypi.org/project/flake8-comprehensions/ + "C90", # check for complexity; https://pypi.org/project/mccabe/ + "D", # docstring style checker; https://pypi.org/project/pydocstyle/ + "DOC", # docstring linter; https://pypi.org/project/pydoclint/ + "E", # style guide checking; https://pypi.org/project/pycodestyle/ + "F", # check for errors; https://pypi.org/project/pyflakes/ + "RUF", # ruff-specific linting rules; https://docs.astral.sh/ruff/rules/#ruff-specific-rules-ruf + "FA", # use from __future__ import annotations if needed; https://pypi.org/project/flake8-future-annotations/ + "FURB", # refurbish and modernize Python codebases; https://pypi.org/project/refurb/ + "I", # sorting imports rules; https://pypi.org/project/isort/ + "N", # check naming conventions; https://pypi.org/project/pep8-naming/ + "PERF", # check performance anti-patterns; https://pypi.org/project/perflint/ + "PT", # check common style issues or inconsistencies in pytest; https://pypi.org/project/flake8-pytest-style/ + "PTH", # use pathlib where possible; https://pypi.org/project/flake8-use-pathlib/ + "S", # security testing; https://pypi.org/project/flake8-bandit/ + "SIM", # check for code that can be simplified; https://pypi.org/project/flake8_simplify/ + "T20", # do not use prints in production; https://pypi.org/project/flake8-print/ + "TRY", # prevent Exception handling anti-patterns; https://pypi.org/project/tryceratops/ + "UP" # upgrade syntax for newer versions; https://pypi.org/project/pyupgrade +] +ignore = [ + "D100", # allow undocumented public module definitions + "D101", # allow undocumented public class + "D203", # do not require one blank line before class docstring + "D213", # first docstring line should be on the second line + "TRY003", # allow exception messages outside the `Exception` class + "F401", # allow unused imports +] + +[lint.per-file-ignores] +"tests/*" = [ + "D101", # allow undocumented public class + "D102", # allow undocumented class method + "D103", # allow undocumented public method definitions + "S101", # allow use of `assert` in test files +] diff --git a/main/tests/__init__.py b/tests/__init__.py similarity index 100% rename from main/tests/__init__.py rename to tests/__init__.py diff --git a/main/tests/conftest.py b/tests/conftest.py similarity index 100% rename from main/tests/conftest.py rename to tests/conftest.py diff --git a/main/tests/fixtures/__init__.py b/tests/fixtures/__init__.py similarity index 100% rename from main/tests/fixtures/__init__.py rename to tests/fixtures/__init__.py diff --git a/main/tests/fixtures/fixture_ftp_server.py b/tests/fixtures/fixture_ftp_server.py similarity index 72% rename from main/tests/fixtures/fixture_ftp_server.py rename to tests/fixtures/fixture_ftp_server.py index 0a0b64ac..5c5ae958 100644 --- a/main/tests/fixtures/fixture_ftp_server.py +++ b/tests/fixtures/fixture_ftp_server.py @@ -1,12 +1,10 @@ -import os +from pathlib import Path + import pytest @pytest.fixture(scope="module") def ftp_file_names() -> list[str]: - """ - This function returns a list of file names for the FTP server - """ return ["file_one.raw", "file_two.raw", "file_three.raw", "config.txt", "README.md"] @@ -16,16 +14,15 @@ def fixture_ftp_server(ftpserver, ftp_file_names): # This will not upload real files, it will only create files that we can collect with the Reader class # From: https://github.com/oz123/pytest-localftpserver/blob/c1ebb0068f4197fad5e14e9c1ef3488de5cf877f/tests/test_pytest_localftpserver.py#L362 - # Reset temporary directories ftpserver.reset_tmp_dirs() # Get the base file path to "upload" files - base_path = ftpserver.get_local_base_path() + base_path = Path(ftpserver.get_local_base_path()) # Define a list of files to create on the local ftp server - files_on_server: list[str] = [] + files_on_server: list[Path] = [] for filename in ftp_file_names: - abs_file_path = os.path.join(base_path, filename) + abs_file_path: Path = base_path / filename files_on_server.append(abs_file_path) - with open(abs_file_path, "w") as f: + with abs_file_path.open("w") as f: f.write(filename) diff --git a/main/tests/inputs/test_async_bioservices_input.csv b/tests/inputs/test_async_bioservices_input.csv similarity index 100% rename from main/tests/inputs/test_async_bioservices_input.csv rename to tests/inputs/test_async_bioservices_input.csv diff --git a/main/tests/inputs/test_proteomics_input.csv b/tests/inputs/test_proteomics_input.csv similarity index 100% rename from main/tests/inputs/test_proteomics_input.csv rename to tests/inputs/test_proteomics_input.csv diff --git a/main/tests/test_model_creation.py b/tests/test_model_creation.py similarity index 75% rename from main/tests/test_model_creation.py rename to tests/test_model_creation.py index 77959ef9..ab2a0396 100644 --- a/main/tests/test_model_creation.py +++ b/tests/test_model_creation.py @@ -1,8 +1,3 @@ -""" -The purpose of this file is to test the result of creating a constraint-based metabolic model with the provided naiveB-cell data -""" - - def test_correct_bracket(): pass diff --git a/main/tests/test_project.py b/tests/test_project.py similarity index 60% rename from main/tests/test_project.py rename to tests/test_project.py index d0724772..3a278bfe 100644 --- a/main/tests/test_project.py +++ b/tests/test_project.py @@ -1,9 +1,5 @@ -import os from pathlib import Path -# Add parent directory to path, allows us to import the "project.py" file from the parent directory -# From: https://stackoverflow.com/a/30536516/13885200 - from como import project @@ -14,4 +10,3 @@ def test_config(): assert configs.data_dir == root_dir / "data" assert configs.config_dir == root_dir / "data" / "config_sheets" assert configs.result_dir == root_dir / "data" / "results" - assert configs.code_dir == root_dir / "como" diff --git a/main/tests/test_proteomics.py b/tests/test_proteomics.py similarity index 81% rename from main/tests/test_proteomics.py rename to tests/test_proteomics.py index a402db43..d3c6ad66 100644 --- a/main/tests/test_proteomics.py +++ b/tests/test_proteomics.py @@ -1,7 +1,3 @@ -""" -This file tests the proteomics module -""" - from pathlib import Path import aioftp @@ -11,8 +7,7 @@ from como.proteomics.FTPManager import Download, Reader, aioftp_client from como.proteomics.proteomics_preprocess import ParseCSVInput, PopulateInformation -from tests.fixtures.fixture_ftp_server import fixture_ftp_server -from tests.fixtures.fixture_ftp_server import ftp_file_names +from tests.fixtures.fixture_ftp_server import fixture_ftp_server, ftp_file_names class TestCrux: @@ -21,23 +16,20 @@ class TestCrux: class TestFileInformation: def test_creating_instances(self, tmp_path: str): - """ - This is an example instance. If anything is incorrectly changed in the FileInformation class, this test will fail. - """ - + """Fail if FileInformation changes.""" # Define several constants cell_type: str = "t_gondii" study: str = "S1" replicate: str = "R1" file_name: str = "2DE_2DE_10-2D" - + # Define paths; this file was chosen because it is only 14MB in size download_url: str = f"ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2015/02/PXD000297/{file_name}.raw" raw_path: Path = Path(tmp_path, f"{cell_type}_{study}{replicate}_{file_name}.raw") mzml_path: Path = Path(tmp_path, f"{cell_type}_{study}{replicate}_{file_name}.mzml") sqt_path: Path = Path(tmp_path, f"{cell_type}_{study}{replicate}_{file_name}.target.sqt") intensity_path: Path = Path(tmp_path, f"protein_abundance_matrix_{cell_type}.csv") - + # Create an instance of FileInformation instance: FileInformation = FileInformation( cell_type=cell_type, @@ -47,12 +39,12 @@ def test_creating_instances(self, tmp_path: str): intensity_csv=intensity_path, mzml_path=mzml_path, sqt_path=sqt_path, - file_size=0 + file_size=0, ) - + # Set the replicate (`R1`) instance.set_replicate(replicate) - + # Check that the instance is correct assert instance.cell_type == cell_type assert instance.download_url == download_url @@ -69,20 +61,23 @@ def test_creating_instances(self, tmp_path: str): class TestFTPManager: @pytest.mark.asyncio async def test_ftp_client(self): - """ - This test checks that the ftp_client function works as expected - """ + """Checks that ftp_client function works as expected.""" host: str = "ftp.pride.ebi.ac.uk" port: int = 21 - client: aioftp.Client = await aioftp_client(host=host, port=port, username="anonymous", password="guest") - + client: aioftp.Client = await aioftp_client( + host=host, + port=port, + username="anonymous", + password="guest", # noqa: S106 + ) + assert client.server_host == host assert client.server_port == port assert await client.get_current_directory() == Path("/") assert await client.quit() is None - + @pytest.mark.skip(reason="pyftpdlib is broken, no way to test this") - def test_reader(self, ftpserver, fixture_ftp_server, ftp_file_names): + def test_reader(self, ftpserver, fixture_ftp_server, ftp_file_names): # noqa: F811 # Use pytest_localftpserver and fixtures.fixture_ftp_server.fix # Now we can get login information for our local FTP server file_extensions: list[str] = ["raw"] @@ -91,26 +86,18 @@ def test_reader(self, ftpserver, fixture_ftp_server, ftp_file_names): port: int = dict_login_data["port"] user: str = dict_login_data["user"] passwd: str = dict_login_data["passwd"] - + # Now we can use our reader class and collect files from the local FTP server - reader = Reader( - file_extensions=file_extensions, - root_link=login_data, - port=port, - user=user, - passwd=passwd - ) - + reader = Reader(file_extensions=file_extensions, root_link=login_data, port=port, user=user, passwd=passwd) + # Assert that we have files ending in "raw", and that the file name found is in the ftp file names list for file in reader.file_names: assert file.endswith(tuple(file_extensions)) assert file in ftp_file_names - + @pytest.mark.skip(reason="pyftpdlib is broken, no way to test this") def test_download(self, mocker, tmp_path): - """ - This checks that the Download class works as expected - """ + """Checks that Download class works as expected.""" class TestProteomicsPreprocess: diff --git a/main/tests/test_rnaseq_preprocess.py b/tests/test_rnaseq_preprocess.py similarity index 60% rename from main/tests/test_rnaseq_preprocess.py rename to tests/test_rnaseq_preprocess.py index 1db53996..752ec333 100644 --- a/main/tests/test_rnaseq_preprocess.py +++ b/tests/test_rnaseq_preprocess.py @@ -1,38 +1,40 @@ -import pytest -import os +from __future__ import annotations + import sys -# Add parent directory to path, allows us to import the "project.py" file from the parent directory -# From: https://stackoverflow.com/a/30536516/13885200 -sys.path.append(os.path.dirname(os.path.dirname(os.path.abspath(__file__)))) -import rnaseq_preprocess +import pytest +from como import rnaseq_preprocess # Define a list of arguments to test -# fmt: off @pytest.mark.skip("Not testing args because the input 'argv' has been removed") @pytest.mark.parametrize( "args", [ # Test using data in COMO_input data ["--context-names", "naiveB immNK", "--gene-format", "Ensembl", "--taxon-id", "9606", "--create-matrix"], - ["--context-names", "dimNK brightNK", "--gene-format", "SYMBOL", "--taxon-id", "human", "--provide-matrix", "--matrix", "COMO_input/counts_matrix.tsv"], + [ + "--context-names", + "dimNK brightNK", + "--gene-format", + "SYMBOL", + "--taxon-id", + "human", + "--provide-matrix", + "--matrix", + "COMO_input/counts_matrix.tsv", + ], ], ) -# fmt: on - def test_arg_input(args: list[str]): - """ - This function asserts that the arguments passed into the function are correct - """ + """Asserts that the arguments passed into the function 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