This project is focused on validation of mouse and human cell culture models of transgenerational epigenetic inheritance (TEI) of environmentally induced epimutations. Our mouse and human cell culture models have shown key advantages toward discovering molecular mechanisms underlying the initial induction and subsequent propagation of epimutations in a rapid, cost-effective, non-animal intense, high-resolution manner. We propose experiments to validate the accuracy with which these models recapitulate in vitro the effects of environmental exposures of intact animals in vivo. We will address the following questions:
1) Can cell culture models replace/reduce the need for live animal experiments to elucidate molecular mechanisms by which adverse environmental effects disrupt the epigenome, predisposing disease states in multiple subsequent generations, even with no further exposure to the initial disruption?
2) Can cell culture models accurately recapitulate induced epimutagenesis seen in intact animals?
3) Are genomic patterns of induced epimutations and differentially expressed genes similar in intact animals and cultured cells?
4) Are induced epimutations retained, erased, or replaced during transitions in cell fate?
We will use comprehensive multiomic single-cell transcriptomic and epigenomic profiling of cell types exposed to disruptors in vivo and in vitro. Our in vitro approach is necessary because 20 years of studies of intact animals have failed to elucidate molecular mechanisms underlying TEI. We seek to validate our cell culture models as a high-resolution alternative to intact animals for future studies. The impact of our experiments will be validation of an in vitro approach that will affect a paradigm shift in the manner in which long-standing fundamental questions are addressed regarding molecular mechanisms by which organisms are impacted by their environment, and how these effects can be transmitted across multiple generations.
