Glucocorticoid receptor epigenetic programming in alcohol-induced neuropathology

项目总结。 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并存。有几条证据表明 慢性酒精暴露对认知功能减退的发展和进展具有有害影响 痴呆症，包括阿尔茨海默病(AD)。酒精诱导的表观遗传学改变(例如，DNA高/低- 甲基化)对基因表达产生长期影响。然而，转录改变的表观遗传学基础 由AUD引起的，它们对认知损害的影响，以及AD神经病理的发展 不清楚。酒精刺激糖皮质激素的释放，过量会扰乱神经功能 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，因为它有助于 糖皮质激素介导的神经病理过程中的关键分子机制。在这里，我们 建议研究糖皮质激素受体(GR)介导的慢性前列腺癌的分子机制 酒精导致转录改变，包括参与AD发展的基因网络，在 酒精依赖的动物模型。我们将使用高通量测序技术(剪切和运行以及 Atac-seq)和通过表观遗传编辑(CRISPR/dCas9)的表观遗传表达操纵来评估酒精- 诱导GR结合的变化以及随后的转录和行为变化(K99期)，以进一步 深入研究认知衰退和神经元病理发展的生物学过程 在老化过程中(R00阶段)。本项目的创新性与杰出的研究成果相结合 环境将帮助申请者获得尖端分子和生物信息学方法的培训，并 作为一名独立的酒精研究人员取得成功所需的额外职业发展机会。Dr。 Eleonora Gatta将接受UIC的一个由资深酒精研究人员和神经科学家组成的团队的培训。这个 导师团队由S.C.Pandey博士(主要导师)、A.Guidotti博士、E.J.格洛弗、A.Lasek和M. Maienschein-Cline将在实验设计、酒精依赖的动物模型、完成 以及对大数据集以及酒精研究中的专业和职业发展的分析。此外, 培训计划包括相当多的赠款撰写、口头陈述、道德规范以及建筑方面的培训。 对Gatta博士在R00期间作为一名独立研究人员的成功至关重要 相位。除了由指导团队提供的培训外，Gatta博士还可以访问 UIC精神病学系和酒精表观遗传学研究中心(CARE)。总而言之， 这个K99/R00应用程序将通过揭示AUD和AD共同发病的新的大脑机制 认知功能减退和AD风险增加的表观遗传学机制 AUD患者的神经病理学。

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.