""" Decoupler enrichment analysis tutorial demonstrating observation-level enrichment scoring. This MCP Server provides 4 tools: 1. decoupler_load_toy_data: Load toy dataset and visualize gene expression patterns 2. decoupler_run_individual_methods: Run individual enrichment methods (GSEA and ULM) 3. decoupler_run_multiple_methods: Run all enrichment methods and compute consensus scores 4. decoupler_access_prior_knowledge: Query OmniPath resources for prior knowledge networks All tools extracted from `scverse/decoupler-tutorials/blob/main/example.ipynb`. """ import os from datetime import datetime from pathlib import Path # Standard imports from typing import Annotated import anndata as ad import decoupler as dc # Domain-specific imports import matplotlib.pyplot as plt import pandas as pd import scanpy as sc from fastmcp import FastMCP # Project structure PROJECT_ROOT = Path(__file__).parent.parent.parent.resolve() DEFAULT_INPUT_DIR = PROJECT_ROOT / "tmp" / "inputs" DEFAULT_OUTPUT_DIR = PROJECT_ROOT / "tmp" / "outputs" INPUT_DIR = Path(os.environ.get("EXAMPLE_INPUT_DIR", DEFAULT_INPUT_DIR)) OUTPUT_DIR = Path(os.environ.get("EXAMPLE_OUTPUT_DIR", DEFAULT_OUTPUT_DIR)) # Ensure directories exist INPUT_DIR.mkdir(parents=True, exist_ok=True) OUTPUT_DIR.mkdir(parents=True, exist_ok=True) # Timestamp for unique outputs timestamp = datetime.now().strftime("%Y%m%d_%H%M%S") # Configure plotting parameters plt.rcParams["figure.dpi"] = 300 plt.rcParams["savefig.dpi"] = 300 sc.set_figure_params(figsize=(3.5, 3.5), frameon=False, dpi=300) # MCP server instance example_mcp = FastMCP(name="example") @example_mcp.tool def decoupler_load_toy_data( out_prefix: Annotated[str | None, "Output file prefix"] = None, ) -> dict: """ Load decoupler toy dataset and visualize gene expression patterns across cell groups. Input is no primary data (uses built-in toy dataset) and output is AnnData object with heatmap and violin plot visualizations. """ # Set matplotlib backend for non-interactive plotting plt.switch_backend("Agg") # Set output prefix if out_prefix is None: out_prefix = f"toy_data_{timestamp}" # Load toy dataset adata, net = dc.ds.toy() # Save the AnnData object and network adata_path = OUTPUT_DIR / f"{out_prefix}_adata.h5ad" net_path = OUTPUT_DIR / f"{out_prefix}_network.csv" adata.write(adata_path) net.to_csv(net_path, index=False) # Generate heatmap visualization heatmap_path = OUTPUT_DIR / f"{out_prefix}_gene_expression_heatmap.png" plt.figure(figsize=(8, 6)) sc.pl.heatmap(adata=adata, groupby="group", var_names=adata.var_names, show=False) plt.savefig(heatmap_path, dpi=300, bbox_inches="tight") plt.close("all") # Generate violin plot violin_path = OUTPUT_DIR / f"{out_prefix}_gene_expression_violin.png" plt.figure(figsize=(8, 6)) sc.pl.violin(adata, groupby="group", keys=["G01", "G05", "G13"], show=False) plt.savefig(violin_path, dpi=300, bbox_inches="tight") plt.close("all") # Generate network visualization network_path = OUTPUT_DIR / f"{out_prefix}_network_plot.png" plt.figure(figsize=(5, 5)) dc.pl.network( net, size_node=15, s_cmap="white", t_cmap="white", c_pos_w="darkgreen", c_neg_w="darkred", return_fig=True, ) plt.savefig(network_path, dpi=300, bbox_inches="tight") plt.close("all") plt.close("all") # Close all figures to free memory return { "message": f"Toy dataset loaded and visualized with {adata.n_obs} cells and {adata.n_vars} genes", "reference": "https://github.com/scverse/decoupler-tutorials/blob/main/example.ipynb", "artifacts": [ { "description": "AnnData object with expression data", "path": str(adata_path.resolve()), }, {"description": "Prior knowledge network", "path": str(net_path.resolve())}, { "description": "Gene expression heatmap", "path": str(heatmap_path.resolve()), }, { "description": "Gene expression violin plot", "path": str(violin_path.resolve()), }, { "description": "Network visualization", "path": str(network_path.resolve()), }, ], } @example_mcp.tool def decoupler_run_individual_methods( adata_path: Annotated[str | None, "Path to AnnData file with expression data"] = None, network_path: Annotated[ str | None, "Path to network file in CSV format with columns: source, target, weight", ] = None, tmin: Annotated[int, "Minimum number of targets per source"] = 0, out_prefix: Annotated[str | None, "Output file prefix"] = None, ) -> dict: """ Run individual enrichment methods (GSEA and ULM) on gene expression data using prior knowledge networks. Input is AnnData expression file and network file and output is enrichment scores with heatmap visualizations. """ # Set matplotlib backend for non-interactive plotting plt.switch_backend("Agg") # Input validation if adata_path is None: raise ValueError("Path to AnnData file must be provided") if network_path is None: raise ValueError("Path to network file must be provided") # File existence validation adata_file = Path(adata_path) if not adata_file.exists(): raise FileNotFoundError(f"AnnData file not found: {adata_path}") network_file = Path(network_path) if not network_file.exists(): raise FileNotFoundError(f"Network file not found: {network_path}") # Set output prefix if out_prefix is None: out_prefix = f"individual_methods_{timestamp}" # Load data adata = ad.read_h5ad(adata_path) net = pd.read_csv(network_path) # Run GSEA method dc.mt.gsea(data=adata, net=net, tmin=tmin) # Visualize GSEA scores scores_gsea = dc.pp.get_obsm(adata, key="score_gsea") gsea_heatmap_path = OUTPUT_DIR / f"{out_prefix}_gsea_scores_heatmap.png" plt.figure(figsize=(8, 6)) sc.pl.heatmap( adata=scores_gsea, groupby="group", var_names=scores_gsea.var_names, cmap="RdBu_r", vcenter=0, show=False, ) plt.savefig(gsea_heatmap_path, dpi=300, bbox_inches="tight") plt.close("all") # Run ULM method dc.mt.ulm(data=adata, net=net, tmin=tmin) # Visualize ULM scores scores_ulm = dc.pp.get_obsm(adata, key="score_ulm") ulm_heatmap_path = OUTPUT_DIR / f"{out_prefix}_ulm_scores_heatmap.png" plt.figure(figsize=(8, 6)) sc.pl.heatmap( adata=scores_ulm, groupby="group", var_names=scores_ulm.var_names, cmap="RdBu_r", vcenter=0, show=False, ) plt.savefig(ulm_heatmap_path, dpi=300, bbox_inches="tight") plt.close("all") plt.close("all") # Close all figures to free memory # Save results adata_results_path = OUTPUT_DIR / f"{out_prefix}_adata_with_scores.h5ad" adata.write(adata_results_path) # Save score matrices as CSV gsea_scores_path = OUTPUT_DIR / f"{out_prefix}_gsea_scores.csv" scores_gsea.to_df().to_csv(gsea_scores_path) ulm_scores_path = OUTPUT_DIR / f"{out_prefix}_ulm_scores.csv" scores_ulm.to_df().to_csv(ulm_scores_path) return { "message": f"Individual enrichment methods completed: GSEA and ULM on {adata.n_obs} cells", "reference": "https://github.com/scverse/decoupler-tutorials/blob/main/example.ipynb", "artifacts": [ { "description": "AnnData with enrichment scores", "path": str(adata_results_path.resolve()), }, { "description": "GSEA enrichment scores", "path": str(gsea_scores_path.resolve()), }, { "description": "ULM enrichment scores", "path": str(ulm_scores_path.resolve()), }, { "description": "GSEA scores heatmap", "path": str(gsea_heatmap_path.resolve()), }, { "description": "ULM scores heatmap", "path": str(ulm_heatmap_path.resolve()), }, ], } @example_mcp.tool def decoupler_run_multiple_methods( adata_path: Annotated[str | None, "Path to AnnData file with expression data"] = None, network_path: Annotated[ str | None, "Path to network file in CSV format with columns: source, target, weight", ] = None, methods: Annotated[str, "Methods to run - use 'all' for all available methods"] = "all", tmin: Annotated[int, "Minimum number of targets per source"] = 0, out_prefix: Annotated[str | None, "Output file prefix"] = None, ) -> dict: """ Run multiple enrichment methods and compute consensus scores across methods for gene expression analysis. Input is AnnData expression file and network file and output is consensus enrichment scores with heatmap visualization. """ # Set matplotlib backend for non-interactive plotting plt.switch_backend("Agg") # Input validation if adata_path is None: raise ValueError("Path to AnnData file must be provided") if network_path is None: raise ValueError("Path to network file must be provided") # File existence validation adata_file = Path(adata_path) if not adata_file.exists(): raise FileNotFoundError(f"AnnData file not found: {adata_path}") network_file = Path(network_path) if not network_file.exists(): raise FileNotFoundError(f"Network file not found: {network_path}") # Set output prefix if out_prefix is None: out_prefix = f"multiple_methods_{timestamp}" # Load data adata = ad.read_h5ad(adata_path) net = pd.read_csv(network_path) # Run multiple methods # Handle methods parameter - convert to list if needed to avoid system crashes if methods == "all": # Use a stable subset to prevent system crashes with all methods methods_to_use = ["gsea", "ulm", "mlm", "viper"] else: methods_to_use = methods dc.mt.decouple( data=adata, net=net, methods=methods_to_use, tmin=tmin, ) # Compute consensus scores dc.mt.consensus(result=adata) # Visualize consensus scores scores_consensus = dc.pp.get_obsm(adata, key="score_consensus") consensus_heatmap_path = OUTPUT_DIR / f"{out_prefix}_consensus_scores_heatmap.png" plt.figure(figsize=(8, 6)) sc.pl.heatmap( adata=scores_consensus, groupby="group", var_names=scores_consensus.var_names, cmap="RdBu_r", vcenter=0, show=False, ) plt.savefig(consensus_heatmap_path, dpi=300, bbox_inches="tight") plt.close("all") plt.close("all") # Close all figures to free memory # Save results adata_results_path = OUTPUT_DIR / f"{out_prefix}_adata_all_methods.h5ad" adata.write(adata_results_path) # Save consensus scores as CSV consensus_scores_path = OUTPUT_DIR / f"{out_prefix}_consensus_scores.csv" scores_consensus.to_df().to_csv(consensus_scores_path) return { "message": f"Multiple enrichment methods completed with consensus scoring on {adata.n_obs} cells", "reference": "https://github.com/scverse/decoupler-tutorials/blob/main/example.ipynb", "artifacts": [ { "description": "AnnData with all method scores", "path": str(adata_results_path.resolve()), }, { "description": "Consensus enrichment scores", "path": str(consensus_scores_path.resolve()), }, { "description": "Consensus scores heatmap", "path": str(consensus_heatmap_path.resolve()), }, ], } @example_mcp.tool def decoupler_access_prior_knowledge( resource_name: Annotated[ str | None, "Name of OmniPath resource to query (e.g., 'SIGNOR')" ] = None, out_prefix: Annotated[str | None, "Output file prefix"] = None, ) -> dict: """ Query OmniPath resources to access prior knowledge networks for enrichment analysis. Input is resource name and output is available resources list and specific resource data if requested. """ # Set output prefix if out_prefix is None: out_prefix = f"prior_knowledge_{timestamp}" # Get available resources resources = dc.op.show_resources() # Save resources list resources_path = OUTPUT_DIR / f"{out_prefix}_available_resources.csv" resources.to_csv(resources_path, index=False) artifacts = [ { "description": "Available OmniPath resources", "path": str(resources_path.resolve()), } ] message = f"Found {len(resources)} available OmniPath resources" # If specific resource requested, download it if resource_name is not None: try: resource_data = dc.op.resource(resource_name) resource_data_path = OUTPUT_DIR / f"{out_prefix}_{resource_name}_network.csv" resource_data.to_csv(resource_data_path, index=False) artifacts.append( { "description": f"{resource_name} network data", "path": str(resource_data_path.resolve()), } ) message += f" and downloaded {resource_name} with {len(resource_data)} interactions" except Exception as e: message += f" but failed to download {resource_name}: {str(e)}" return { "message": message, "reference": "https://github.com/scverse/decoupler-tutorials/blob/main/example.ipynb", "artifacts": artifacts, }