RegNetAgents: A Multi-Agent Framework for Cross-Network Regulatory Driver Identification in Cancer Genomics

We introduce RegNetAgents, an AI-oriented multi-agent framework for structured, query-driven regulatory candidate identification across heterogeneous gene regulatory networks. The system enables unified analysis of bulk tumor and single-cell-derived ARACNe networks by integrating TCGA-derived cancer networks with large-scale single-cell regulatory networks from the GREmLN project. For a given focal gene, the framework performs dual-network classification, cancer gene filtering using OncoKB annotations, and mode-of-action (MoA) assignment for tumor-derived regulatory relationships. Candidates are ranked by evidence consistency across networks (Both, TCGA-only, GREmLN-only). The system is implemented as a multi-agent LangGraph DAG workflow, accessible through a unified Python API and Model Context Protocol (MCP) client, operating as a downstream analytical layer over precomputed regulatory networks rather than a network inference method. Across eleven breast cancer (BRCA) and twelve colorectal cancer (COAD) focal genes, RegNetAgents identifies candidate regulators significantly enriched for OncoKB-annotated cancer genes. TCGA-derived candidates show strong enrichment (Stouffer Z = 6.69 for BRCA and 6.95 for COAD), while GREmLN-derived candidates also demonstrate significant enrichment (Z = 5.51 for BRCA and 7.06 for COAD; all p < 0.0001). No enrichment is observed in housekeeping or non-driver control gene sets, supporting signal specificity. An extended module enables structured evaluation of oncogenic potential, druggability, clinical relevance, and network vulnerability, supporting end-to-end interpretation from candidate identification to biological hypothesis generation. RegNetAgents establishes an interpretable AI framework for cross-network regulatory candidate identification in cancer genomics.