Systems-Level Transcriptomics Maps Multilevel Remodeling and Pathway-Selective Translational Alignment Across Murine Models of Cardiometabolic HFpEF

Heart failure with preserved ejection fraction (HFpEF) is a heterogeneous cardiometabolic syndrome in which the molecular programs linking metabolic stress to myocardial remodeling and diastolic dysfunction remain incompletely defined. We integrated ventricular RNA sequencing with pathway activity profiling, transcription factor inference, cell-type enrichment, phenotype association, elastic-net severity modeling, cross-lab murine validation, and human proteomic comparison to define the systems-level architecture of remodeling in the db/db + aldosterone mouse model of cardiometabolic HFpEF. HFpEF hearts exhibited a distinct transcriptomic state characterized by coordinated upregulation of collagen organization, TGFβ signaling, inflammatory response, and NFκB signaling, with reduced ion-channel activity and smaller shifts in oxidative phosphorylation, excitation–contraction coupling, and mechanotransduction. These pathway programs were linked to left ventricular hypertrophy and diastolic dysfunction and were accompanied by enrichment of fibroblast, myofibroblast, and macrophage signatures that tracked the same disease dimensions. Gene-level prioritization identified extracellular matrix, inflammatory, and mechanotransduction-associated candidates linked to disease severity, while transcription factor analysis revealed a broader multi-regulator architecture associated with fibrotic, inflammatory, and stress-responsive remodeling. Elastic-net modeling further showed that phenotype-derived remodeling severity was captured in an exploratory nested cross-validation framework primarily by transcription factor and fibro-inflammatory cell-program features, whereas pathway-summary scores added little incremental predictive information. In an independent HFD+L-NAME cohort, pathway remodeling showed selective reproducibility, and cross-species comparison demonstrated that concordance with human HFpEF proteomic subgroups was pathway selective rather than global. Together, these findings define a multilevel systems architecture of cardiometabolic HFpEF remodeling and support mechanistic prioritization and pathway-matched preclinical model selection.