Glucocorticoid receptor epigenetic programming in alcohol-induced neuropathology

项目摘要。 酒精使用障碍（AUD）是一种复杂的大脑疾病，其特征是一系列持续的行为和 神经化学表现常常与认知障碍共病。几条证据表明， 慢性酒精暴露对认知能力下降的发展和进展具有有害影响， 痴呆，包括阿尔茨海默病（AD）。酒精诱导的表观遗传改变（例如，DNA高/低 甲基化）对基因表达产生长期影响。然而，转录组变化的表观遗传基础 其对认知障碍的影响以及AD神经病理学的发展仍然存在 不清楚酒精会刺激糖皮质激素的释放，过量的糖皮质激素会破坏神经功能 并导致与年龄相关的认知能力下降。因此，酒精会增加认知负担， 糖皮质激素介导的神经病理学的关键分子机制。这里我们 建议检查糖皮质激素受体（GR）介导的分子机制，通过这些机制，慢性 酒精产生转录组学变化，包括参与AD发展的基因网络， 酒精依赖动物模型我们将使用高通量测序技术（CUT&RUN和 ATAC-seq）和通过表观遗传编辑（CRISPR/dCas 9）进行的表观遗传表达操作，以评估酒精- 诱导GR结合的变化以及随后的转录组学和行为变化（K99期），以进一步 深入研究认知衰退和神经病理学发展的生物学过程 老化期间（R 00阶段）。该项目的创新性质与杰出的研究相结合， 环境将帮助申请人获得尖端分子和生物信息学方法的培训， 额外的职业发展机会，作为一个独立的酒精研究人员取得成功所必需的。博士 Eleonora Gatta将接受UIC高级酒精研究人员和神经科学家团队的培训。的 导师团队，由S.C.博士组成。Pandey（primary mentor），A.格洛弗，A. Lasek和M. Maienschein-Cline将提供关于实验设计，酒精依赖动物模型，完成 和大数据集的分析以及酒精研究的专业和职业发展。此外，本发明还提供了一种方法， 培训计划包含了大量的关于赠款写作、口头陈述、道德和建筑方面的培训 生物信息学管道，这将是至关重要的博士加塔的成功，作为一个独立的研究人员在R 00 相位除了指导小组提供的培训外，Gatta博士还将获得 UIC精神病学系和表观遗传学酒精研究中心（CARE）。总的来说， 这项K99/R 00应用将通过揭示 认知能力下降易感性增加和AD风险增加的表观遗传机制 在患有AUD的个体中的神经病理学

PROJECT SUMMARY. Alcohol use disorder (AUD) is a complex brain disorder characterized by an array of persistent behavioral and neurochemical manifestations often comorbid with cognitive impairments. Several lines of evidence indicate that chronic alcohol exposure has a deleterious effect on the development and progression of cognitive decline and dementia, including Alzheimer’s disease (AD). Alcohol-induced epigenetic alterations (e.g., DNA hyper/hypo- methylation) exert long-lasting effects on gene expression. Yet, the epigenetic basis for transcriptomic changes resulting from AUD, their impact on cognitive impairments, and the development of AD neuropathology remains unclear. Alcohol stimulates the release of glucocorticoids, which in excessive amounts, disrupt neuronal function and contribute to age-related cognitive decline. Thus, alcohol augments the cognitive burden by contributing to key molecular mechanisms underlying neuropathology via glucocorticoid-mediated processes. Here, we propose to examine the glucocorticoid receptor (GR)-mediated molecular mechanisms through which chronic alcohol produces transcriptomic changes, including gene networks involved in the development of AD, in an animal model of alcohol dependence. We will use high-throughput sequencing techniques (CUT&RUN and ATAC-seq) and epigenetic expression manipulation via epigenetic editing (CRISPR/dCas9) to assess alcohol- induced changes in GR binding and consequent transcriptomic and behavioral changes (K99 phase) to further delve into the biological processes underlying the development of cognitive decline and neuronal pathology during aging (R00 phase). The innovative nature of this project combined with the outstanding research environment will help the applicant gain training in cutting-edge molecular and bioinformatic approaches and additional career development opportunities necessary for success as an independent alcohol researcher. Dr. Eleonora Gatta will receive training from a team of senior alcohol researchers and neuroscientists at UIC. The mentorship team, composed of Drs. S.C. Pandey (primary mentor), A. Guidotti, E.J. Glover, A. Lasek and M. Maienschein-Cline, will offer insights on experiment design, animal model of alcohol dependence, completion and analyses of big data sets as well as on professional and career development in alcohol research. In addition, the training plan contains considerable training on grant writing, oral presentations, ethics, as well as construction of bioinformatic pipelines that will be crucial to Dr. Gatta’s success as an independent researcher during the R00 phase. In addition to training provided by the mentoring team, Dr. Gatta will have access to resources in the Department of Psychiatry at UIC and in the Center for Alcohol Research on Epigenetics (CARE). Collectively, this K99/R00 application will identify novel brain mechanisms in the co-morbidity of AUD and AD by uncovering epigenetic mechanisms underlying increased susceptibility for cognitive decline and risk to develop AD neuropathology in individuals with AUD.