Multiomics analysis of concurrent MASH and atherosclerosis in male and female mice

Metabolic dysfunction-associated steatohepatitis (MASH) has reached epidemic proportions with limited therapies available. Surprisingly, the leading cause of death in these patients is atherosclerotic cardiovascular disease. No pharmacological therapy exists to treat MASH and atherosclerosis simultaneously, which is attributed to the lack of translational mouse models, particularly in females. Herein, we have utilized both male and female atheroprone mice, combined with different high-fat diets containing physiologically relevant cholesterol levels, to model concurrent MASH-atherosclerosis. Upon phenotypic characterization using standard histopathology, immunohistochemistry, and biochemical assays on liver, cardiovascular, and plasma samples, we report the first female MASH-atherosclerosis model. Furthermore, we conducted a multi-omics exploration of metabolomic, lipidomic, and transcriptomic profiles altered within these groups, and found metabolic and genomic signatures comparable to the human disease. These models will facilitate future cardiometabolic research to unravel the molecular and metabolic underpinnings at the intersection of MASH and atherosclerosis, which must be pharmacologically targeted. Overall design: Eight-week-old Ldlr-/- male and female mice were housed under controlled temperature (22±2°C) and humidity conditions (40-60%) on a 12-hour light/dark cycle and fed ad libitum a standard chow diet (LabDiet #5053, 13% of calories from fat). For our dietary model we used established29,42 high-fat and high-cholesterol western diet (WD, Envigo TD.88137, 42 kcal% fat with 0.2% cholesterol), choline deficient with high fat and high cholesterol diet (Research Diets #D22052704, 45 kcal% fat without added choline and 0.2% cholesterol) and modified MASH diet (Research Diets #D22052703, 40 kcal% fat with palmitic acid and trans-fat levels and 0.2% cholesterol) for 24 weeks prior to euthanasia and tissue harvest.