Multiple Generation Distinct Toxicant Exposures Induce Epigenetic-Transgenerational-Inheritance of Enhanced Pathology and Obesity

This study examines the actions of the agricultural fungicide vinclozolin on the F0 generation, followed by jet fuel hydrocarbon mixture exposure of the F1 generation, and then pesticide DDT (dichlorodiphenyltrichloroethane) on the F2 generation gestating females. The subsequent F3 and F4 generations and F5 transgenerational-generation were examined for male sperm epigenetic alterations and pathology in males and females. Significant impacts on the male sperm differential DNA methylation regions (DMRs) were observed. The F3-F5 generations were similar in approximately 50% of the DMRs. The pathology of each generation was assessed in the testis, ovary, kidney, prostate, obesity, and tumors. The pathology used a newly developed Deep Learning (DL), artificial intelligence (AI), based histopathology analysis. Observations demonstrated compounded disease impacts in obesity and metabolic parameters, but other pathologies plateaued with smaller increases at the F5 transgenerational generation. Therefore, the multiple generational exposures, which occurs in human populations, may increase epigenetic impacts and disease susceptibility. The experimental design used three successive generations of gestating female exposures during fetal gonadal sex determination, followed by three generations of no exposure to obtain the F5 generation to reveal the ultimate transgenerational phenotype for pathology and disease. The F0 generation gestating female was exposed to the agricultural fungicide vinclozolin. The second successive exposure of the F1 generation gestating female involved jet fuel (JP8). The third successive exposure of the F2 generation gestating female involved DDT (dichlorodiphenyltrichloroethane). The subsequent F3 and F4 generations and F5 transgenerational-generation were examined for male sperm epigenetic alterations and pathology in males and females. Methylation differences between treatment groups was done using an MeDIP-seq procedure. The control lineage was only taken to the F3 generation. This F3 generation control was then compared with the F3, F4, and F5 treatment lineage sample groups.