Reliable interpretability of biology-inspired deep neural networks

This code supplements the publication by Esser-Skala and Fortelny (2023).

Folders

(Not all of these folders are included in the git repository.)

• data: output files generated by P-NET and DTox (available from https://doi.org/10.5281/zenodo.7760561)
• doc: project documentation
• docker: files for creating Docker containers with P-NET or DTox installed
• literature: relevant publications
• plots: generated plots
• pnet_data: P-NET data files
• renv: R environment data
• scripts: bash and R scripts

Preparation

P-NET and DTox

Download the provided Docker containers from the GitHub Container registry:

bash docker pull ghcr.io/csbg/pnet-container:1.0.0 docker pull ghcr.io/csbg/dtox-container:1.0.0

Alternatively, pull these containers with Apptainer/Singularity:

bash singularity pull docker://ghcr.io/csbg/pnet-container:1.0.0 singularity pull docker://ghcr.io/csbg/dtox-container:1.0.0

When using the latter container format, replace all calls to run_[pnet/dtox]_docker.sh with run_[pnet/dtox]_singularity.sh.

R

In order to run the R scripts, you will need one of the following:

• an installation of R 4.3.1; restore required packages from renv.lock via bash Rscript -e "renv::restore()"`

• the Docker container available from the GitHub Container registry: bash docker pull ghcr.io/csbg/r_pnet_robustness:1.0.0 Replace calls to Rscript below by scripts/run_rscript_docker.sh.

• the Apptainer/Singularity container: bash singularity pull docker://ghcr.io/csbg/r_pnet_robustness:1.0.0 Replace calls to Rscript below by scripts/run_rscript_singularity.sh.

Datasets

Download the MSK-IMPACT 2017 dataset:

bash wget https://cbioportal-datahub.s3.amazonaws.com/msk_impact_2017.tar.gz tar xzf msk_impact_2017.tar.gz -C pnet_data

Run P-NET experiments

Generally, each experiment comprises the following steps:

1. Load P-NET input data via load_data_[dataset].R.
2. Optionally, modify input data via modify_data_[technique].R.
3. Run P-NET via Docker using the provided bash script run_pnet_docker.sh. This script has three arguments:
   • -e experiment: experiment name, required
   • -l [n]: lower seed, optional (default: -1, which uses the original seeds)
   • -u [n]: upper seed, optional (default: 49)

Within each experiment, results from each run are saved in a subfolder indicating the two random seeds used (e.g., data/pnet_original/0_0).

utils.R is required by all data preparation scripts.

Original setup

Run P-NET with the original setup as described in the publication.

bash Rscript scripts/load_data_original.R scripts/run_pnet_docker.sh -e pnet_original

Deterministic inputs

Input data is modified so that presence of mutation and copy number amplification is perfectly correlated with class label 1 (copy number deletion is always 0).

bash Rscript scripts/load_data_original.R Rscript scripts/modify_data_deterministic.R scripts/run_pnet_docker.sh -e pnet_deterministic

Shuffled labels

Shuffle training/test labels before each run using uniform class frequencies.

bash for seed in {-1..49}; do Rscript scripts/load_data_original.R Rscript scripts/modify_data_shuffled.R FALSE $seed scripts/run_pnet_docker.sh -e pnet_shuffled_each -l $seed -u $seed done

MSK-IMPACT 2017 dataset

```bash Rscript scripts/loaddatamskimpact.R "Non-Small Cell Lung Cancer" scripts/runpnetdocker.sh -e mskimpactnsclcoriginal

for seed in {-1..49}; do Rscript scripts/loaddatamskimpact.R "Non-Small Cell Lung Cancer" Rscript scripts/modifydatashuffled.R FALSE $seed scripts/runpnetdocker.sh -e mskimpactnsclcshuffled -l $seed -u $seed done

Rscript scripts/loaddatamskimpact.R "Breast Cancer" scripts/runpnetdocker.sh -e mskimpactbcoriginal

for seed in {-1..49}; do Rscript scripts/loaddatamskimpact.R "Breast Cancer" Rscript scripts/modifydatashuffled.R FALSE $seed scripts/runpnetdocker.sh -e mskimpactbcshuffled -l $seed -u $seed done

Rscript scripts/loaddatamskimpact.R "Colorectal Cancer" scripts/runpnetdocker.sh -e mskimpactccoriginal

for seed in {-1..49}; do Rscript scripts/loaddatamskimpact.R "Colorectal Cancer" Rscript scripts/modifydatashuffled.R FALSE $seed scripts/runpnetdocker.sh -e mskimpactccshuffled -l $seed -u $seed done

Rscript scripts/loaddatamskimpact.R "Prostate Cancer" scripts/runpnetdocker.sh -e mskimpactpcoriginal

for seed in {-1..49}; do Rscript scripts/loaddatamskimpact.R "Prostate Cancer" Rscript scripts/modifydatashuffled.R FALSE $seed scripts/runpnetdocker.sh -e mskimpactpcshuffled -l $seed -u $seed done ```

Run DTox experiments

Run DTox with seeds ranging from 0 (i.e., the original seed) to 50.

bash scripts/run_dtox_docker.sh

Results from each run are saved in a subfolder indicating the random seed used (e.g., data/dtox/0).

Analyze results

plot_figures.R generates all figures shown in the publication, using files in data (described below):

bash Rscript scripts/plot_figures.R

styling.R is required by this script.

P-NET

After each run, the following files are copied from the P-NET output folders:

• analysis/extracted/node_importance_graph_adjusted.csv (renamed to node_importance.csv): contains node importance scores, with the following columns:
  • (first, unnamed): node name
  • coef: original node importance scores
  • coef_graph: indegree plus outdegree of node
  • coefcombined: adjusted node importance score (= coef / coefgraph if coefgraph > mean(coefgraph) + 5 sd(coef_graph) in the respective layer)
  • coefcombinedzscore: scaled coef_combined
  • coefcombined2: z(z(coefgraph) - z(coef))
  • layer: layer of the node
• _logs/p1000/pnet/onsplit_average_reg_10_tanh_large_testing/P-net_ALL_testing.csv (renamed to predictions_test.csv): predictions for the test set, with the following columns:
  • (first, unnamed): sample name
  • pred: predicted class (unfortunately, encoded by a double 1.0 or 0.0)
  • pred_scores: probability of the predicted class
  • y: true class (encoded as integer 1 or 0)
• _logs/p1000/pnet/onsplit_average_reg_10_tanh_large_testing/P-net_ALL_training.csv (renamed to predictions_train.csv): predictions for the training set (same columns as above)

DTox

The following files generated by DTox are required for subsequent analyses:

• module_relevance.tsv: contains node importance scores, with the following columns:
  • (first, unnamed): compound identifier
  • remaining columns: node identifiers (UniProt and Reactome IDs)
• test_labels.csv: predictions for the test set, with two columns:
  • truth: true label (0 or 1)
  • predicted: predicted label (decimal number between 0 and 1)

Appendix: How to build Docker images

P-NET

The folder docker/pnet contains everything needed for building a Docker image with P-NET installed:

• Dockerfile: instructions for assembling the image
• entrypoint.sh: script for running P-NET; used as entrypoint in the container
• environment_pnet.yml: conda environment specification
• patch_seeds.diff: patch that allows to change the random seed for P-NET
• setup.sh: executed during image assembly; installs P-NET (with input data) and conda

Build and deploy this image via

bash docker build --tag ghcr.io/csbg/pnet-container:1.0.0 . docker push ghcr.io/csbg/pnet-container:1.0.0

DTox

The folder docker/dtox contains everything needed for building a Docker image with DTox installed:

• Dockerfile: instructions for assembling the image
• entrypoint.sh: entrypoint in the container; activates conda environment
• environment_dtox.yml: conda environment specification
• patch_seeds.diff: patch that allows to change the random seed for DTox and saves predicted labels for the test set
• run_dtox.py: executes the DTox workflow as described in the tutorial available in the DTox GitHub repository
• setup.sh: executed during image assembly; installs DTox and conda

Build and deploy this image via

bash docker build --tag ghcr.io/csbg/dtox-container:1.0.0 . docker push ghcr.io/csbg/dtox-container:1.0.0

R

The folder docker/r contains the Dockerfile needed for building a Docker image with R and required packages installed.

Build and deploy this image via

bash cp ../../renv.lock . docker build --tag ghcr.io/csbg/r_pnet_robustness:1.0.0 . docker push ghcr.io/csbg/r_pnet_robustness:1.0.0

Appendix: Description of files required by P-NET

• genes/: only the genes present in both of the following two files will be analyzed: (a) tcga_prostate_expressed_genes_and_cancer_genes.csv (b) HUGO_genes/protein-coding_gene_with_coordinate_minimal.txt (TSV, no column names; meaning of columns: chromosome, start, end, gene name)
• pathways/:
  • pathways_short_names.xlsx: short pathway names for figure labels
  • Reactome/ReactomePathways.gmt: genes associated with Reactome pathways; TSV, no column names, variable number of columns: (1) pathway name (2) reactome id (3) type (unused) (4ff) associated genes
  • Reactome/ReactomePathways.txt: TSV mapping Reactome ids to names; loaded by P-NET but apparently not used (?)
  • Reactome/ReactomePathwaysRelation.txt: TSV specifying the Reactome pathway hierarchy as edge list; columns indicate parent and child; only human pathways are used (i.e., the child id has to start with "HSA")
• prostate/
  • processed/
  • P1000_final_analysis_set_cross_important_only.csv: mutation data; first column contains sample name, remaining columns represent genes, cells indicate number of mutations; data is preprocessed to a binary matrix, indicating presence/absence of at least one mutation (i.e., 1 if original >= 1)
  • P1000_data_CNA_paper.csv: CNV data; first column (unnamed) contains sample name, remaining columns represent genes, cells indicate copy number status; data is preprocessed to two binary matrices: one indicates presence of copy number amplification (1 if original > 1.5), the other indicates presence of CN deletion (1 if original < -1.5)
  • response_paper.csv: input labels, two columns: (1) id – sample name (2) response – sample label (1 = metastatic tumor)
  • splits/: splits of input data; all files have three columns: (1) [unnamed] – running number starting at zero (2) id – sample name (3) response – sample label (column is NOT used by P-NET!)
  • test_set.csv: samples in the test set
  • training_set_0.csv: samples in the training set
  • validation_set.csv: samples in the validation set