Neuroepigenetic mechanisms of alcohol response - PROJECT SUMMARY The overarching goal of the research presented in this application is to reveal the fundamental and conserved genetic and epigenetic programs that encode and transmit a memory of alcohol exposure for several generations. In mammals, in utero ethanol exposure is associated with an array of well-characterized neurobehavioral issues. However, there is mounting evidence in a variety of model organisms that some adverse neurological features are also detectable in the third generation following exposure, indicating a transgenerational effect. Alcohol also has a clear epigenetic impact and directly contributes to the modification of the epigenome. Nevertheless, it is unclear how the memory of ethanol exposure persists in the nervous system across generations. Here, I combine the tractability and conservation of the model system Caenorhabditis elegans with state-of-the-art epigenomic analyses, classical genetics, and behavioral approaches to shed light on the mechanisms of epigenetic memory of alcohol exposure and its transgenerational behavioral effects. My preliminary data shows that ethanol exposure (1) leads to increased histone acetylation in directly exposed animals; and that (2) ethanol seeking behavior is increased until at least the third generation, paralleling the increased alcohol consumption observed in rodent models and humans. Whether these observations are connected, and ethanol-mediated acetylation is directly responsible for the altered transgenerational impact on behavior is unknown. The specific altered acetylation marks and their distribution across the neuronal genome are also unknown. Thus, the driving hypothesis of this project is that neuronal histone hyperacetylation persists across generations to drive ethanol-induced transgenerational behavioral effects. In Aim 1, I will use a histone multiplex assay, immunofluorescence, and CUT&RUN sequencing of FACS-sorted neuronal nuclei to identify the neuronal epigenetic changes accompanying exposure to ethanol. In Aim 2, I will pharmacologically inhibit and perform neuron subtype-specific RNA interference of histone acetyltransferases and demethylases to determine the role of neuronal histone acetylation in ethanol’s transgenerational behavioral effects. This proposal has the potential to establish the impact of ethanol exposure on neuronal histone PTMs and their role in transgenerational behavioral effects. By applying genetic, epigenetic, and pharmacological approaches in a relevant and highly tractable organism, this proposed work will advance our knowledge of the epigenetic memory of alcohol exposure and its transgenerational behavioral effects. I will carry out these studies in Patrick Allard’s lab at UCLA, a respected expert in environmental epigenetics, with support from co-sponsors Xia Yang, an expert in epigenomics, and Kelly Huffman, an expert in fetal alcohol spectrum disorders. The environment at UCLA will also provide excellent intellectual, technical, and professional training. This will aid me in achieving my career goal of running an independent research lab studying the neuroepigenetics of chemical exposures.
