Sex-based multiomics analysis uncovers metabolic and molecular mediators linking MASH and atherosclerosis

Atherosclerotic cardiovascular disease (ASCVD) is the leading cause of death in patients with metabolic dysfunction-associated steatohepatitis (MASH). No therapy targets both diseases simultaneously, and a roadblock for discovering new treatments is the lack of animal models that recapitulate both diseases, especially in females.

Male and female Ldlr ^-/- mice (n = 8-13) were fed a western diet (WD), modified choline-deficient high-fat diet (mCDHFD), or modified MASH-inducing diet (mMASHD) containing equivalent physiological levels of cholesterol. Comprehensive multiomics including metabolomics, lipidomics, and transcriptomics, alongside histopathological and biochemical analyses, were integrated to characterize concurrent MASH and atherosclerosis. Transcriptomics was validated in other mouse models and integrated with human data (n = 79).

While mCDHFD induced MASH-fibrosis in both sexes, WD was effective only in males, whereas mMASHD primarily affected females. mCDHFD induced concurrent MASH and atherosclerosis in both sexes, while WD effectively recapitulated disease co-occurrence only in males. Correlation analyses highlighted links between MASH and atherosclerosis, identifying circulating cholesterol and C-C motif chemokine ligand 2 (CCL2) as potential predictors of coexisting disease (p <0.04). Integrated metabolomic and transcriptomic analyses identified arginine-proline, glycine-serine, glutathione, and sphingolipid metabolism (p <0.03) as key dysregulated pathways, with sphinganine emerging as a predictor of disease severity. Hepatic itaconate and lactate levels were positively correlated with disease severity, whereas glycine, carnitine, 2-aminomuconic acid, and thiamine pyrophosphate were negatively associated (p <0.04). Lipidomic analyses revealed dysregulated polyunsaturated fatty acid, steryl ester, and dihexosylceramide metabolism. Integration of mouse and human transcriptomes revealed similarities in metabolic and proinflammatory/proatherogenic pathways.

This sex-based multiomics analysis establishes a murine model of concurrent MASH and atherosclerosis, reveals sex-specific dietary responses, and identifies metabolic and transcriptional pathways with potential utility as biomarkers and therapeutic targets.

This study addresses the critical need for an animal model that replicates both metabolic dysfunction-associated steatohepatitis (MASH) and atherosclerotic cardiovascular disease, particularly in females, to facilitate therapeutic development. Using male and female Ldlr ^-/- mice, we found that different diets containing equivalent physiological levels of cholesterol induce sex-specific responses, with a modified choline-deficient high-fat diet effectively modeling both diseases in both sexes, while a western diet is effective only in males. Multiomics analyses identified key metabolic and inflammatory pathways linking MASH and atherosclerosis that mirror transcriptomic signatures found in humans, and highlight circulating cholesterol, CCL2, and sphinganine as potential biomarkers. These findings establish a translational model and reveal sex-specific metabolic pathways that will advance our understanding of the shared pathophysiology of MASH and atherosclerosis, and facilitate the development of dual therapeutic approaches, addressing an urgent unmet clinical need.