Mitochondrial NNT Promotes Diastolic Dysfunction in Cardiometabolic HFpEF

BACKGROUND: Clinical management of heart failure with preserved ejection fraction (HFpEF) is hindered by a lack of disease-modifying therapies capable of altering its distinct pathophysiology. Despite the widespread implementation of a 2-hit model of cardiometabolic HFpEF to inform precision therapy, which utilizes ad libitum high-fat diet and 0.5% N(ω)-nitro-L-arginine methyl ester, we observe that C57BL6/J mice exhibit less cardiac diastolic dysfunction in response to high-fat diet and 0.5% N(ω)-nitro-L-arginine methyl ester. METHODS: Genetic strain-specific single-nucleus transcriptomic analysis identified disease-relevant genes that enrich oxidative metabolic pathways within cardiomyocytes. Because C57BL/6J mice are known to harbor a loss-of-function mutation affecting the inner mitochondrial membrane protein Nnt (nicotinamide nucleotide transhydrogenase), we used an isogenic model of Nnt loss-of-function to determine whether intact NNT is necessary for the pathological cardiac manifestations of high-fat diet and 0.5% N(ω)-nitro-L-arginine methyl ester. Twelve-week-old mice cross-bred to isolate wild-type ( Nnt +/+ ) or loss-of-function ( Nnt − / − ) Nnt in the C57BL/6N background were challenged with high-fat diet and 0.5% N(ω)-nitro-L-arginine methyl ester for 9 weeks (n=6–10). RESULTS: Nnt +/+ mice exhibited impaired ventricular diastolic relaxation and pathological remodeling, as assessed via E/e′ (42.8 versus 21.5, P =1.2×10 − 10 ), E/A (2.3 versus 1.4, P =4.1×10 − 2 ), diastolic stiffness (0.09 versus 0.04 mm Hg/μL, P =5.1×10 − 3 ), and myocardial fibrosis ( P =2.3×10 − 2 ). Liquid chromatography and mass spectroscopy exposed a 40.0% reduction in NAD + ( P =8.4×10 − 3 ) and a 38.8% reduction in glutathione:GSSG ( P =2.6×10 − 2 ) among Nnt +/+ mice after high-fat diet and 0.5% N(ω)-nitro-L-arginine methyl ester feeding. Using single-nucleus ligand-receptor analysis, we implicate Fgf1 (fibroblast growth factor 1) as a putative NNT-dependent mediator of cardiomyocyte-to-fibroblast signaling of myocardial fibrosis. CONCLUSIONS: Together, these findings underscore the pivotal role of mitochondrial dysfunction in HFpEF pathogenesis, implicating both NNT and Fgf1 as novel therapeutic targets.