Establishing a human cellular model of sex differences in the brain

众所周知，生物性别会影响许多精神疾病的风险、发病年龄和/或严重程度。 以及神经系统疾病。男性患帕金森氏症、自闭症和精神分裂症的几率更高 而女性患严重抑郁障碍、焦虑症、阿尔茨海默病和 多发性硬化症。此外，研究人员还观察到神经系统属性上的性别差异 与神经发生、形态、基因表达、突触密度和连接性有关。这些差异 可能是由于差异遗传(XX和XY)、荷尔蒙影响和/或社会的复杂影响 环境。要了解影响疾病和“正常”大脑功能的具体因素， 这对于理解个体间这种变异的细胞和分子基础至关重要。尽管有一些 引人注目的动物模型工作，研究分析人类性别差异的细胞和分子基础 直到最近，大脑还很少见。部分原因是缺乏人体细胞模型。 神经科学的系统，既与生物学有关，又受基因控制。在此，我们建议 建立一个可控的、明确的、可操作的人类干细胞模型，用于研究 遗传性行为对神经精神疾病的细胞机制的影响。建议的系统将提供 美国和其他国家开发了一种便捷的“插件”系统，用于在基于干细胞的分析中添加性别差异成分。在……里面 目的1，我们建议建立XX和XY干细胞系，它们在性染色体之外是完全等基因的 互补性。性别差异不能完全归因于遗传学，就像常染色体基因组 也是不同的，而且由于不存在自然出现的常染色体同基因的男性/女性对， 我们建议在这里设计出基因相同但性别不同的人类干细胞系。 染色体互补。我们将使用我们手中的Klinefelter胚胎干细胞系(XXY)和 从Klinefelter成纤维细胞开发新的IPSCs。然后，我们将诱导这些行丢失单个X或Y 染色体，创造常染色体男性和女性干细胞系的亚克隆。在目标2中，我们将使用 在SA1中建立常染色体等基因干细胞系统以识别差异的基因和蛋白质 受基因性别的影响。为此，我们将把我们的同基因XX和XY干细胞分别分化为 下丘脑、大脑皮层、星形胶质细胞和小胶质细胞的神经元及其差异效应 XY和XX细胞在基因和蛋白质调控上的差异。

It is well established that biological sex affects risk, age-of-onset, and/or severity of many psychiatric and neurological diseases. Men have a higher incidence of Parkinson's disease, autism, and schizophrenia while women show a higher incidence of major depressive disorder, anxiety disorder, Alzheimer's disease and multiple sclerosis. Additionally, researchers have observed sex differences across nervous system properties related to neurogenesis, morphology, gene expression, synapse density, and connectivity. These differences could be due to differential genetics (XX vs XY), hormonal influence, and/or the complex effects of social environment. To understand the specific factors influencing both disease and “normal” brain function, it is critical to understand the cell and molecular bases of this variation between individuals. In spite of some compelling animal model work, studies analyzing the cell and molecular bases of sex differences in the human brain have, until very recently, been rare. Part of the reason for this is the lack of a human cellular model system for neuroscience that is both biologically relevant and genetically controlled. Here, we propose to establish a well controlled, defined, and manipulatable human stem cell model for studying the impact of genetic sex on cellular mechanisms underlying neuropsychiatric diseases. The system proposed would provide us and others a facile “plug-in” system for adding a sex differences component to stem cell-based analyses. In Aim 1, we propose to develop XX and XY stem cell lines that are fully isogenic outside of the sex chromosome complement. Sex differences cannot be definitively attributed to genetics as long as the autosomal genome also is also divergent, and since there is no natural occurrence of an autosomally isogenic male/female pair, we propose here to engineer human stem cell lines that are genetically identical with the exception of the sex chromosome complement. We will use a Klinefelter embryonic stem cell line (XXY) that we have in hand and develop new iPSCs from Klinefelter fibroblasts. We then will induce these lines to lose either a single X or Y chromosome, creating subclones of autosomally male and female stem cell lines. In Aim 2, we will use the autosomally isogenic stem cell system developed in SA1 to identify genes and proteins that are differentially affected by genetic sex. For this aim we will differentiate our isogenic XX and XY stem cells individually into neurons of the hypothalamus and of the cortex, astrocytes, and microglia and assay differential effects between XY and XX cells on gene and protein regulation.