Understanding Cell-type Vulnerability and Oxidative Stress Pathology in Parkinson's Disease Using Isogenic Human Dopaminergic Neurons

项目总结：了解细胞类型的脆弱性和操纵氧化应激病理 用同基因人多巴胺能神经元治疗帕金森病 大约有100万美国人患有帕金森氏病(PD)，其特征是进行性丧失 黑质中脑多巴胺能神经元亚群，导致运动障碍和其他 令人衰弱的条件。家族性帕金森病基因在脑内广泛表达，帕金森病患者神经退行性变 然而，与其他细胞相比，为什么黑质dns表现出如此微妙的脆弱性尚不清楚。 类型，包括其他目录号码人群。尸检研究表明，氧化应激(OS)导致 家族性和散发性帕金森病。活性氧簇(ROS)是重要的信号分子，但 细胞内ROS会损伤DNA、脂质和蛋白质。高能量需求和多巴胺代谢可能 解释增加的ROS、OS和黑名单域名系统的独特漏洞，但与人类相关的模型系统 需要对这一假设进行严格的检验。迫切需要开发实验系统，以更好地 了解黑质糖尿病肾病的脆弱性，识别与疾病相关的新信号机制，并改进分子 分型和患者分层。在本补充资料中，我们建议使用我们开发的最新模型 并将其应用于剖析病理细胞类型的特定机制。我们最近开发了3D文化 整合诱导多能干细胞(IPSCs)衍生的单个细胞类型(神经元，星形胶质细胞， 少突胶质细胞，内皮细胞来创造脑组织芯片。这种方法允许我们将细胞与 不同的遗传背景。为此，我们的目标是研究家族性帕金森病的细胞类型特异性影响 基因Parkin和DJ1，以更好地了解每种脑细胞类型在帕金森病病理中所起的作用。 具体地说，我们的目标是确定每种细胞类型中的Parkin变体对人类氧化应激的影响 通过创建具有帕金功能丧失的不同细胞类型突变的脑组织芯片。我们会 监测多巴胺能神经元的氧化应激反应。此外，我们的目标是确定DJ1风险的影响 通过引入不同的细胞类型，每种细胞类型中的变异对蛋白平衡和多巴胺能神经元存活的影响 DJ1功能变异缺失。我们将测量DJ1缺失所导致的蛋白分解活性和神经元死亡。 在不同的细胞类型中的功能。将3D组织芯片系统应用于PD将使我们能够识别 每种细胞类型在帕金森病的病理中起重要作用。

Project Summary: Understanding cell-type vulnerability and manipulating oxidative stress pathology in Parkinson’s Disease using isogenic human dopaminergic neurons About one million Americans live with Parkinson’s Disease (PD) which is characterized by progressive loss of subpopulations of nigral midbrain dopaminergic neurons (DNs), leading to motor impairment and other debilitating conditions. Familial PD genes show broad expression in the brain and neurodegeneration in PD can be widespread; however, it is unclear why nigral DNs show such exquisite vulnerability compared to other cell types, including other DN populations. Post-mortem studies suggest that oxidative stress (OS) contributes to familial and sporadic PD. Reactive oxygen species (ROS) are important signaling molecules but high levels of intracellular ROS will damage DNA, lipids and proteins. High energy needs and dopamine metabolism may explain increased ROS, OS and the unique vulnerability of nigral DNs but human-relevant model systems are required to rigorously test this hypothesis. There is an urgent need to develop experimental systems to better understand nigral DN vulnerability, identify novel disease-relevant signaling mechanisms, and improve molecular subtyping and patient stratification. In this supplement we propose to use a recent model that we have developed and apply it to dissect pathological cell-type-specific mechanisms. We have recently developed a 3D culture model integrating induced pluripotent stem cells (iPSCs) derived individual cell types (neurons, astrocytes, oligodendrocytes, endothelial cells to create brain tissue chips. This approach allows us to integrate cells with different genetic backgrounds. To this end, we aim to investigate the cell type specific effects of familial PD genes PARKIN and DJ1 to better understand the role each brain cell type plays to contribute to PD pathology. Specifically, we aim to determine the impact of PARKIN variants in each cell type on oxidative stress in human brain tissue by creating brain tissue chips with PARKIN loss of function mutations in different cell types. We will monitor oxidative stress in the dopaminergic neurons. Additionally, we aim to determine the impact of DJ1 risk variants in each cell type on proteostasis and dopaminergic neuron survival by introducing different cell types with DJ1 loss of function variants. We will measure proteolytic activity and neuronal death as a result of DJ1 loss of function in different cell types. Applying the 3D tissue chip system to PD will allow us to identify the role of each cell type in contributing to PD pathology.