X chromosome inactivation in sex disparities to substance use disorder

项目摘要/摘要 与物质滥用障碍相关的性别差异，临床前和临床前和 临床研究。女性一般更容易受到自闭症的启动、升级和戒断的影响 物质滥用行为多于男性。尽管这些差异很大程度上归因于荷尔蒙 不同的是，非激素因素调节成瘾的证据已经被一些 学习。然而，性染色体影响物质滥用行为的潜在机制 代表着我们在物质使用障碍的表观遗传学方面的知识基础上的巨大差距。我们提议写一部小说 女性逃脱X染色体失活(XCI)有助于性行为的假说 成瘾行为的差异。我们将应用尖端技术和独特的新方法 我们最近开发的工具来全面调查XCI逃脱对性相关的影响 上瘾的差异。XCI是一种发生在哺乳动物雌性身上的表观遗传机制，它的作用是 使性别间的基因表达均衡。女性有两条X染色体(XX)，在XCI期间，一条X染色体 随机选择的染色体被转录沉默。然而，已知的是，许多X关联 基因逃避XCI，表现为双等位基因表达。这项建议的目标是确定 XCI逃避对物质滥用障碍性别差异的贡献。首先，我们将使用小说 研究细胞单等位基因(XCI)或双等位基因(XCI逸出)基因表达的尖端小鼠模型 与阿片类药物和精神刺激剂成瘾相关的特定X连锁基因：单胺氧化酶A (MAOA)和GABAA受体A3(Gabra3)。我开创了一种创新的方法，使用了一种基因特异的双重双链 顺反子报告鼠作为可视化这些成瘾相关基因的等位基因使用的工具 在体内的成瘾模型中。接下来，我们将确定XCI在脑组织中的分子图谱和特异性 长期接触阿片类药物和精神刺激剂期间的神经细胞，使用高度创新的单细胞 RNA测序技术。为了实现这些目标，我组建了一支才华横溢的多学科团队， 成瘾、神经科学、遗传老鼠模型、生物信息学和生物统计学的研究合作者。 这种创新的方法来研究和分析基因特异性XCI逃逸作为一种表观遗传机制 物质滥用的背景有可能开辟一个新的研究领域 上瘾。此外，这些转基因小鼠可用于研究XCI逃逸作为表观遗传机制 在其他神经系统疾病中。作为一名早期研究人员，这些研究也将促进我的长期工作 目标是成为物质使用障碍表观遗传学的未来领导者。

PROJECT SUMMARY/ABSTRACT There is a sex-based disparity associated with substance abuse disorders, which is evidenced by preclinical and clinical studies. Females are generally more vulnerable to the initiation, escalation and withdrawal effects of substance abuse behavior than males. Although these differences have largely been attributed to hormonal differences, evidence for non-hormonal factors that regulate addiction has been demonstrated by a number of studies. However, the mechanisms underlying sex chromosome influences on substance abuse behavior represent a huge gap in our knowledge base on the epigenetics of substance use disorders. We propose a novel hypothesis that escape from X-chromosome inactivation (XCI) in females contributes to sex associated differences in addiction behavior. We will apply cutting edge technology and uniquely novel approaches and tools we developed recently to comprehensively investigate the impact of XCI escape on sex associated disparities in addiction. XCI is an epigenetic mechanism that occurs in mammalian females and serves to equalize gene expression between the sexes. Females have two X chromosomes (XX), and during XCI, one X chromosome is randomly chosen to be transcriptionally silenced. However, it is known that a number of X linked genes escape XCI and display bi allelic gene expression. The objective of this proposal is to determine the contribution of XCI escape on sex-associated differences in substance abuse disorder. First, we will use novel cutting edge mouse models to characterize cellular mono-allelic (XCI) or bi-allelic (XCI escape) gene expression of specific X-linked genes associated with addiction to opioids and psychostimulants: monoamine oxidase A (Maoa) and GABAA receptor A3 (Gabra3). I pioneered an innovative approach using a gene specific dual bi- cistronic reporter mouse as a tool to enable the visualization of allelic usage of these addiction associated genes in vivo in a model of addiction. Next, we will determine the molecular landscape of XCI in brain tissue and specific neuronal cells during chronic exposure to opioids and psychostimulants, using a highly innovative single cell RNA sequencing technology. To accomplish these goals, I have assembled a talented, multidisciplinary team of research collaborators in addiction, neuroscience, genetic mouse modelling, bioinformatics and biostatistics. This innovative approach to the study and analysis of gene specific XCI escape as an epigenetic mechanism in the context of substance abuse has the potential to open up a new area of research on the epigenetics of addiction. Further, these genetically modified mice can be used to study XCI escape as an epigenetic mechanism in other neurologic disorders. As an early stage investigator, these studies will also advance my long term objective of becoming a future leader in the epigenetics of substance use disorders.