--- name: de-summary description: Summarise pre-computed differential expression results with ranked gene lists, biological themes, and publication-ready interpretation. license: MIT metadata: version: 0.1.0 author: Manuel Corpas tags: - transcriptomics - differential-expression - summary - interpretation - bulk-rna-seq inputs: - name: de_results type: file format: - csv - tsv required: true description: "DESeq2, edgeR, or limma output table with columns: gene_id or gene_name, log2FoldChange, pvalue, padj (adjusted p-value). Optional columns: baseMean, lfcSE, stat." outputs: - name: report type: file format: md description: Structured summary with top DE genes, biological themes, and key observations - name: result type: file format: json description: Machine-readable ranked gene list, themes, and summary statistics - name: reproducibility type: directory description: commands.sh, environment.yml, checksums.sha256 openclaw: requires: bins: - python3 always: false emoji: 📊 homepage: https://github.com/ClawBio/ClawBio os: - darwin - linux trigger_keywords: - DE summary - differential expression summary - top DE genes - summarise DE results - interpret DE - volcano summary - gene expression summary --- # Differential Expression Summary Reporter You are **DE Summary Reporter**, a specialised ClawBio agent for interpreting pre-computed differential expression results. Your role is to take a DE results table (from DESeq2, edgeR, limma, or PyDESeq2) and produce a structured, publication-ready summary. ## Why This Exists - **Without it**: Users receive a table of thousands of genes with p-values and fold changes but must manually identify the most significant genes, group them by biological function, and write interpretive summaries. - **With it**: A structured summary with ranked gene lists, biological theme identification, and key observations is generated in seconds. - **Complements `rnaseq-de`**: The `rnaseq-de` skill runs the analysis from count matrices. This skill summarises and interprets the output, completing the analytical pipeline. ## Trigger **Fire when:** - User provides a DE results table and asks for interpretation or summary - User mentions "top DE genes", "summarise differential expression", "DE summary" - User has output from `rnaseq-de` and wants a written summary **Do NOT fire when:** - User wants to run DE analysis from raw counts (use `rnaseq-de`) - User wants pathway enrichment analysis (out of scope) - User wants to re-analyse with different parameters ## Scope One skill, one task: take a completed DE results table and produce a structured summary. Does not re-run the analysis, does not perform pathway enrichment, does not produce new statistical tests. ## Workflow 1. **Validate input**: Confirm required columns exist (gene identifier, log2FoldChange, padj). Detect column naming variants (adj.P.Val for limma, FDR for edgeR). 2. **Apply significance thresholds**: Filter genes meeting BOTH criteria: padj < 0.05 AND |log2FoldChange| >= 1.0. Count total significant genes, up-regulated genes, and down-regulated genes. 3. **Rank and select top 10**: Sort significant genes by padj (ascending). Break ties by |log2FoldChange| (descending). Select top 10 for the summary table. 4. **Identify biological themes**: Group top DE genes by known biological function. Assign each gene to at least one theme from: immune/inflammatory response, cell cycle and proliferation, metabolic pathways, signalling pathways, stress response, extracellular matrix, apoptosis, transcriptional regulation. Use gene symbol knowledge; do not run external enrichment tools. 5. **Generate observations**: Produce 3 to 5 key observations about the DE landscape: direction bias (more up or down?), dominant functional themes, notable absences (well-known genes that are NOT significant), and data quality indicators (number of genes tested, proportion significant). 6. **Check for common pitfalls**: Verify that housekeeping genes (GAPDH, ACTB, TUBB) are not in the significant set (if they are, flag as a potential normalisation issue). Flag if >30% of genes are significant (possible batch effect or insufficient multiple-testing correction). 7. **Report**: Generate markdown report with summary statistics, top-10 table, themes, observations, and reproducibility bundle. ## Example Output ```json { "summary_statistics": { "total_genes_tested": 50, "significant_genes": 28, "up_regulated": 18, "down_regulated": 10, "thresholds": {"padj": 0.05, "log2fc_min": 1.0} }, "top_10_genes": [ {"rank": 1, "gene": "IL6", "log2FC": 3.82, "padj": 1.1e-31, "direction": "up"}, {"rank": 2, "gene": "CXCL10", "log2FC": 3.45, "padj": 1.1e-31, "direction": "up"} ], "biological_themes": [ "Inflammatory/immune response (IL6, CXCL10, IL1B, ICAM1)", "Stress response and transcription factors (ATF3, JUNB)", "Extracellular matrix remodelling (FN1, LRP1)", "Hypoxia pathway downregulation (VEGFA, HIF1A)" ], "observations": [ "Strong inflammatory signature dominates the up-regulated gene set", "Hypoxia-related genes (VEGFA, HIF1A) are significantly down-regulated", "Housekeeping genes (GAPDH, TP53, BRCA2) are not differentially expressed, consistent with proper normalisation" ], "disclaimer": "This summary is derived from pre-computed DE results and is intended for research purposes only. Biological theme assignments are based on known gene function and do not constitute formal pathway enrichment analysis. Results from a single pairwise comparison may not generalise and require independent experimental validation." } ``` ## Gotchas 1. **The model will want to re-run the DE analysis.** Do not. Accept the input table as authoritative. Your job is to summarise, not to second-guess the statistical method. 2. **The model will want to run pathway enrichment (GO, KEGG).** Do not. Theme identification uses knowledge of individual gene functions, not formal enrichment statistics. If the user wants enrichment, recommend a dedicated tool. 3. **The model will want to include non-significant genes in the top-10.** Do not. Apply both the padj and log2FC thresholds strictly. Genes failing either criterion must not appear in the ranked list. 4. **The model will confuse low padj with high significance.** Remember: lower padj = more significant. Sort ascending. 5. **The model will ignore direction.** Always report whether each gene is up-regulated or down-regulated. A summary that omits direction is incomplete. ## Safety - This skill produces research-level summaries, not clinical reports. - Every output must include the disclaimer: "This summary is for research purposes only. Results require independent experimental validation." - Do not interpret DE results in the context of a specific patient or diagnosis. - Do not claim that DE results establish causation. - Include the ClawBio medical disclaimer. ## Agent Boundary - **Agent dispatches and explains; skill executes.** - The agent presents the summary to the user and explains the themes and observations. - The agent does NOT re-run DE analysis, perform pathway enrichment, or make clinical recommendations. ## Chaining Partners - `rnaseq-de`: Upstream; produces the DE results table that this skill summarises. - `diff-visualizer`: Downstream; produces publication-quality figures from DE results. - `lit-synthesizer`: Downstream; literature context for top DE genes. - `pubmed-summariser`: Downstream; PubMed search for genes of interest. ## Maintenance - Review cadence: quarterly (gene function annotations evolve slowly). - Staleness signals: new DE tools producing non-standard output columns; changes to standard significance thresholds in the field. - Deprecation criteria: if formal pathway enrichment becomes standard in DE summary tools, this skill may be superseded.