Sperm miR-142-3p reprogramming mediates paternal stress-induced non-alcoholic steatohepatitis in offspring rats

Abstract

Abstract Although epidemiological data suggest a strong association between paternal adverse environmental exposure and susceptibility to multiple diseases in offspring, the sperm-to-liver pathway involved in offspring disease is complicated and worthy of further exploration. Caffeine contained in many beverages is regarded as a chronic stressor, and exerts reproductive and developmental toxicity. Effects of paternal pre-pregnant caffeine exposure (PPCE) on the long-term health of offspring and the underlying mechanisms remain unclear. This study innovatively reported the occurrence and transgenerational inheritance of PPCE-induced non-alcoholic steatohepatitis (NASH) in offspring, and aimed to elucidate its sperm reprogramming mechanism and the potential intervention targets. Male rats were administrated with caffeine (15 ~ 60 mg/kg/d) by gavage for 8 weeks before mating. Symptoms of NASH were found in two successive generations of male rats (F1 and F2) whose fathers or grandfathers (F0) were exposed to caffeine. RNA-seq was employed to screen out a novel miRNA mediating equilibrant of liver fatty metabolism: miR-142-3p. Role of sperm miR-142-3p in PPCE-induced offspring NAFLD was validated by in vitro fertilization of the sperm of PPCE or miR-142-3pKO sperm with normal oocytes. Overexpression of miR-142-3p in offspring liver reversed NASH manifestation in PPCE male offspring. We further proved that caffeine-induced paternal chronic stress (high glucocorticoid level) but not caffeine itself is the main cause of methylation changes in sperm and offspring NAFLD, via experiments in vitro and glucocorticoid receptor blockade. Moreover, the linkage between serum high glucocorticoids and sperm miR-142-3p low programming was also verified in clinical samples. Overall, we demonstrated for the first time that PPCE induced NASH in offspring with transgenerational inheritance, confirmed the reprogramming mechanism of sperm miR-142-3p, and identified miR-142-3p as a potential intervention target for paternal-derived NASH.