Mapping a genome-scale in vivo knockout screen to a mechanistic network model identifies VAV2, RASA1, and LEPR as regulators of cardiomyocyte hypertrophy.

Cardiomyocyte hypertrophy is a leading clinical predictor of heart failure, yet newly identified candidate genes often remain disconnected from the signaling mechanisms that govern cardiomyocyte growth. We developed a computational-experimental pipeline that integrates genome-scale mouse knockout phenotypes with a logic-based differential equation model of hypertrophic signaling. Among 9,605 genes evaluated by the International Mouse Phenotyping Consortium, 939 knockout lines induced abnormal heart morphology. Directional curation of hypertrophy-related sub-phenotypes followed by interaction-based network expansion mapped 37 genes to the signaling model. Virtual knockdown screening identified five candidates with concordant in vivo and in silico effects: LRIG1 and CBL as predicted negative regulators and VAV2, RASA1, and LEPR as predicted positive regulators. Mechanistic subnetwork analysis linked these candidates to distinct receptor-proximal, Ras, PI3K-AKT, and MAPK signaling axes. In neonatal rat cardiomyocytes, siRNA-mediated depletion of VAV2, RASA1, or LEPR reduced phenylephrine-induced cell growth, supporting their cell-autonomous contribution to hypertrophy. Quantitative phenotyping further validated the predicted decreased cardiac hypertrophy for VAV2 and LEPR knockouts but identified potential age-dependent mechanisms for RASA1 knockout. Overall, this study establishes the application of network models to translate from in vivo phenotypic screens into pathway mechanisms.