Mechanisms of Manganese Neurotoxicity

描述（由申请人提供）：锰（Mn）是一种强效神经毒素。我们假设PARK2，一个强大的帕金森氏病（PD）遗传风险因素，改变神经元的脆弱性细胞锰状态的修饰剂，特别是在线粒体功能障碍和氧化应激水平。本研究的长期目标是阐明锰诱导的神经毒性的基础，并确定基于机制的神经保护策略，以减轻人类锰暴露风险。我们的方法将利用一种新的高通量测定细胞内锰水平，以确定细胞锰状态和神经毒性的小分子修饰剂。将定义Mn转运和毒性的遗传修饰剂，并将利用基于人诱导多能干细胞（hiPSC）技术的原代人神经元模型系统对现有和新鉴定的Mn毒性遗传和小分子修饰剂进行转化研究。目的1将在体外利用纹状体和中脑小鼠神经元细胞系和体内利用C.优雅目的2将阐明在体内和体外调节锰转运和毒性的功能途径。具体目标3将检验以下假设：PD的人神经元模型对细胞Mn状态的扰动表现出增加的敏感性。这些特定的目标持有的承诺，描绘共同的启动信号的调制锰神经毒性，揭示机制和易感性与暴露于这种金属。这种双PI建议是支持其在不同的模型系统中使用创新的最先进的互补方法。

DESCRIPTION (provided by applicant): Manganese (Mn) is a potent neurotoxin. We hypothesize that PARK2, a strong Parkinson's disease (PD) genetic risk factor, alters neuronal vulnerability to modifiers of cellular Mn status, particularly at the level of mitochondrial dysfunction and oxidative stress. The long-term goal of this research is to elucidate the basis of Mn-induced neurotoxicity and to identify mechanistic-based neuroprotective strategies to mitigate human Mn exposure risk. Our approach will utilize a novel high-throughput assay of intracellular Mn levels to identify small molecule modifiers of cellular Mn status and neurotoxicity. Genetic modifiers of Mn transport and toxicity will be defined and translational studies of existing and newly identified genetic and small molecule modifiers of Mn toxicity will be performed utilizing a primary human neuronal model system based upon human induced pluripotent stem cell (hiPSC) technology. Aim 1 will identify lead compounds that alter neuronal Mn transport and toxicity in vitro using striatal and mesencephalic murine neuronal cell lines and in vivo using C. elegans. Aim 2 will delineate functional pathways that regulate Mn transport and toxicity in vivo and in vitro. Specific Aim 3 will test the hypothesis that human neuronal models of PD exhibit increased sensitivity to perturbations of cellular Mn status. These specific aims hold the promise of delineating common initiator signals for the modulation of Mn neurotoxicity, shedding light on mechanisms and susceptibility associated with exposure to this metal. This dual-PI proposal is bolstered by its use of innovative state-of-the-art complimentary approaches in diverse model systems.