Novel Proteomics Approach to HIV-Associated Neurocognitive Disorder & Drug Abuse

DP 1 DA 041722的AD相关管理补充，“异常蛋白S-亚硝基化介导 基于hiPSC的路易体痴呆（LBD）模型中的突触和神经元毒性” 总结 路易体痴呆（Lewy body dementia，LBD）被归类为阿尔茨海默病（Alzheimer's disease，AD）相关的痴呆，代表了阿尔茨海默病（Alzheimer's disease，AD）相关的痴呆。 在65岁以上的患者中，AD是仅次于AD的第二种最常见的神经退行性疾病。关键 LBD的病理标志是α-突触核蛋白（αSyn）内含物的积累，尽管淀粉样蛋白-β（Aβ） 和tau蛋白的病理也是典型的，这表明在老年人中有共同的途径导致痴呆症。 LBD和AD。事实上，在LBD和AD大脑中，突触损伤和神经元损伤至少部分导致， 过量产生的活性氧和氮物种（ROS/RNS）产生的有毒 αSyn和/或Aβ的亚种。我们是第一个发现异常蛋白质S-亚硝基化（形成 SNO-蛋白），通过RNS（例如，一氧化氮[NO]）的产生，导致突触/神经元损伤。类似于 我们在HIV相关的人类死后大脑中的父母DP 1研究中发现， 认知障碍与甲基苯丙胺药物滥用（手/冰毒），对于这个补充建议，我们 提供了LBD hiPSC模型和LBD人脑中异常S-亚硝基化蛋白的证据。 因此，这一补充申请涉及父DP 1补助金;此外， 这是对我们先前关于AD人iPSC和AD脑的工作的补充，但与我们先前关于AD人iPSC和AD脑的工作完全不同。 获得NIDA DP 1奖。例如，我们的初步数据提供了证据， PINK 1、MEF 2C和Drp 1的肾脏相关蛋白S-亚硝基化在疾病中起致病作用- LBD中的相关损坏。具体地说，我们证明，预防这些异常蛋白质S-亚硝基化， 反应改善疾病过程，在很大程度上消除线粒体功能障碍，突触损伤， 在基于细胞的模型和动物模型中均出现神经元丢失，表现为错误折叠的αSyn。因此，我们建议 本补充申请的具体目的如下： 具体目的1：确定寡聚体αSyn和Aβ对SNO-PINK 1/SNO-MEF 2C/SNO-1的影响。 HiPSC衍生的神经元培养模型中Drp 1介导的突触功能障碍和神经元细胞死亡 LBD。我们将通过CRISPR/Cas9引入不可亚硝基化的突变体PINK 1，MEF 2C或Drp 1，以测试 SNO-PINK 1、SNO-MEF 2C和SNO-Drp 1是否以因果关系的方式促进突触损伤， 神经元细胞死亡重要的是，我们将比较寡聚体αSyn/Aβ对hiPSC衍生的皮质细胞的作用。 神经元与A9多巴胺能神经元，因为LBD可能优先影响大脑皮层而不是黑质， 分别

AD-Related Administrative Supplement for DP1 DA041722, “Aberrant protein S-nitrosylation mediates synaptic and neuronal toxicity in hiPSC-based models of Lewy Body Dementia (LBD)” SUMMARY Lewy body dementia (LBD), classified as an Alzheimer’s disease (AD)-related dementia, represents the second most common form of neurodegenerative disease after AD in patients above 65 years of age. The key pathological hallmark of LBD entails accumulation of a-synuclein (αSyn) inclusions, although amyloid-β (Aβ) and tau pathologies are also typical, suggesting that there are common pathways contributing to dementia in both LBD and AD. In fact, in LBD and AD brains, synaptic injury and neuronal damage result, at least in part, from excessive generation of reactive oxygen and nitrogen species (ROS/RNS) engendered by toxic subspecies of αSyn and/or Aβ. We were the first group to show that aberrant protein S-nitrosylation (forming SNO-proteins), via RNS (e.g., nitric oxide [NO]) generation, results in synaptic/neuronal damage. Similar to our findings in the parent DP1 grant in human postmortem brains from patients with HIV-associated cognitive disorder with methamphetamine drug abuse (HAND/meth), for this supplemental proposal we present evidence for aberrantly S-nitrosylated proteins in LBD hiPSC models and LBD human brain. Hence, this supplemental application relates to the parent DP1 grant; moreover, the new work complements but is totally distinct from our work on AD human iPSCs and AD brain in a prior supplement to the parent NIDA DP1 award. As an example, our preliminary data provide evidence that mitochondrial-related protein S-nitrosylation of PINK1, MEF2C and Drp1 plays a causative role in disease- related damage in LBD. Specifically, we demonstrate that prevention of these aberrant protein S-nitrosylation reactions ameliorates the disease process, largely abrogating mitochondrial dysfunction, synaptic damage, and neuronal loss in both cell-based and animal models manifesting misfolded αSyn. Accordingly, we propose the following Specific Aim for this Supplemental Application: Specific Aim 1: To determine the effects of oligomeric αSyn and Aβ on SNO-PINK1/SNO-MEF2C/SNO- Drp1—mediated synaptic dysfunction and neuronal cell death in hiPSC-derived neuronal culture models of LBD. We will introduce non-nitrosylatable mutant PINK1, MEF2C, or Drp1 via CRISPR/Cas9 in order to test whether SNO-PINK1, SNO-MEF2C, and SNO-Drp1 contribute in a causal manner to synaptic injury and neuronal cell death. Importantly, we will compare the effects of oligomeric αSyn/Aβ on hiPSC-derived cortical neurons vs. A9 dopaminergic neurons since LBD may preferentially affect cerebrocortex over substantia nigra, respectively.