Genetic Analysis of Copper Toxicity Mechanisms in iPSC-derived Human Neurons

描述(申请人提供)：铜是一种对神经元来说是一种强大的有毒氧化剂，其游离水平必须严格控制。它也是神经元酶反应所必需的微量营养素，如神经递质和神经肽的合成。铜对神经细胞的重要性通过Menkes病得到了说明，Menkes病是一种X连锁遗传病，其特征是神经元组织铜饥饿、神经元极性改变和细胞存活表型。这些铜依赖神经表型背后的确切细胞机制仍不清楚，并构成了这一应用的重点。门克斯病中受影响的基因ATP7A是一种高尔基体定位蛋白，可以将铜加载到分泌蛋白中。这一事实表明，门克斯病的表型是由于整个铜敏感的分泌蛋白质组的改变造成的。我们建议通过全面鉴定人类诱导多能干细胞(IPSCs)分化为神经元的人神经元铜敏感分泌蛋白质组来检验这一假设。我们将测试铜敏感蛋白质组在Menkes病神经元病理进展和严重程度中的参与。这些知识将有助于我们理解和发展门克斯病以及受铜供应影响的疾病的治疗方法，如阿尔茨海默病。

DESCRIPTION (provided by applicant): Copper is a powerful toxic oxidant for neurons whose free levels must be tightly controlled. It is also an essential micronutrient necessary for neurona enzymatic reactions, such as neurotransmitter and neuropeptide synthesis. The importance of copper to neuronal cells is illustrated by Menkes disease, an X-linked genetic disorder characterized by neuronal tissue copper starvation, altered neuronal polarity, and cell survival phenotypes. The precise cellular mechanisms underlying these copper- dependent neuronal phenotypes remain unknown and constitute the focus of this application. The gene affected in Menkes disease, ATP7A, is a Golgi localized protein that loads copper into secretory proteins. This fact suggests that Menkes disease phenotypes result from alterations in the entire copper sensitive secreted proteome. We propose to test this hypothesis by comprehensively identifying the human neuronal copper-sensitive secreted proteome in human induced pluripotent stem cells (iPSCs) differentiated into neurons. We will test the participation of the copper-sensitive proteome in the progression and severity of Menkes disease neuronal pathology. Such knowledge will contribute to our understanding and development of therapeutics in Menkes disease as well as diseases affected by copper availability, such as Alzheimer's disease.