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

描述（由申请人提供）：铜是一种对神经元具有强大毒性的氧化剂，其游离水平必须严格控制。它也是神经元酶促反应所必需的必需微量营养素，如神经递质和神经肽的合成。铜对神经元细胞的重要性通过Menkes病说明，Menkes病是一种X连锁遗传疾病，其特征在于神经元组织铜饥饿、改变的神经元极性和细胞存活表型。这些铜依赖性神经元表型背后的精确细胞机制仍然未知，并构成本申请的焦点。门克斯病中受影响的基因ATP 7A是一种高尔基体定位的蛋白质，其将铜装载到分泌蛋白中。这一事实表明，门克斯病表型是由整个铜敏感性分泌蛋白质组的改变引起的。我们建议通过全面鉴定分化为神经元的人诱导多能干细胞（iPSC）中的人神经元铜敏感性分泌蛋白质组来验证这一假设。我们将测试参与铜敏感蛋白质组的进展和严重程度的门克斯病神经病理。这些知识将有助于我们理解和开发门克斯病以及受铜可用性影响的疾病（如阿尔茨海默病）的治疗方法。

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.