Molecular mechanisms of selective vulnerability of neurons to tauopathy

神经元选择性易损性的分子机制

• 批准号: 10667153
• 负责人: CARLO L CONDELLO
• 金额: $ 74.44万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-06-01 至 2028-02-29
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10667153
• 关键词: Affect; Alzheimer' s Disease; Alzheimer' s disease brain; Alzheimer' s disease related dementia; Alzheimer' s disease therapy; Automobile Driving; Autopsy; Brain; Brain region; CRISPR interference; CUL3 gene; CUL5 gene; Cells; Complement; Complex; Dementia; Development; Disease; Disease model; Enzymes; Genes; Goals; Human; In Situ; Induced pluripotent stem cell derived neurons; Kinetics; Metabolism; Methods; Modeling; Molecular; Mus; Neurodegenerative Disorders; Neurons; Pathogenesis; Pathology; Pathway interactions; Patients; Population; Process; Public Health; Research; Role; Stress; Tauopathies; Technology; Testing; Therapeutic; Validation; Work; causal variant; differential expression; entorhinal cortex; experimental study; genome-wide; glucose metabolism; human tissue; in vivo; induced pluripotent stem cell; innovation; knock-down; member; metabolomics; mouse model; neuron loss; new therapeutic target; promote resilience; resilience; resilience factor; single nucleus RNA-sequencing; small molecule; tau Proteins; tau aggregation; therapeutic evaluation; therapeutic target; ubiquitin-protein ligase

PROJECT SUMMARY A major challenge for the development of effective, disease-modifying therapies for Alzheimer’s disease (AD) and related dementias (ADRD) has been our incomplete understanding of the molecular processes controlling pathogenesis. Important clues for the key molecular pathways controlling AD/ADRD pathogenesis are likely to be gained from the study of selective vulnerability of neurons to AD/ADRD. While different factors are likely to contribute to selective vulnerability, our central hypothesis is that cell-autonomous pathways in neurons contribute to selective vulnerability in AD/ADRD, and that these pathways are potential therapeutic targets to reduce neuronal vulnerability to disease. Therefore, there is an urgent need to uncover the neuronal pathways casually driving selective vulnerability, and to test their therapeutic potential. In order to uncover determinants of selective vulnerability in AD, we previously used single-nucleus RNA sequencing to provide the first molecular description of selectively vulnerable neurons in the human entorhinal cortex, a brain region affected early in AD by tau pathology and neuronal loss. Neuronal subtypes that were lost early in disease were also selectively affected by tau pathology. This work provided us with a list of differentially expressed genes between relatively vulnerable versus resilient neuronal populations. The next challenge is to determine which of these differentially expressed genes causally contributes to selective vulnerability. To establish a causal role of specific differentially expressed genes in selective vulnerability, we will leverage CRISPRi technology, which enables the control of expression levels of endogenous genes. CRISPRi was co-developed by MPI Dr. Kampmann, who also pioneered CRISPRi in human iPSC-derived neurons and optimized its use in mouse brains (see preliminary results). In a genome-wide CRISPRi modifier screen in human iPSC-derived neurons, we identified several pathways controlling levels of tau pathology. By comparing hits from the CRISPRi screen to genes that are differentially expressed between resilient and vulnerable neurons in the human AD brain, we uncovered candidate resilience factors, including an CUL5 E3 ubiquitin ligase complex (Aim 1) and candidate vulnerability factors, including key glycolytic enzymes (Aim 2). The goal of Aim 3 is to conduct a large-scale CRISPRi screen for factors controlling tau pathology directly in the brain of tauopathy mouse models. The focus of the proposed project is to uncover mechanisms underlying selective neuronal vulnerability in AD and ADRD. These mechanisms represent potential therapeutic targets, and future research will test the therapeutic potential of targeting the identified pathways. The experimental strategy we propose to establish here to uncover mechanisms underlying selective vulnerability to tauopathy will provide a blueprint that can be applied to many other neurodegenerative diseases.

项目摘要 开发有效的阿尔茨海默病（AD）疾病改善疗法的主要挑战 和相关痴呆症（ADRD）一直是我们对控制ADRD的分子过程的不完全理解。 发病机制控制AD/ADRD发病机制的关键分子途径的重要线索可能是 从AD/ADRD神经元选择性易损性的研究中获得。虽然不同的因素可能会 我们的中心假设是，神经元中的细胞自主通路 有助于AD/ADRD的选择性脆弱性，这些途径是潜在的治疗靶点， 降低神经元对疾病的脆弱性。因此，迫切需要揭示神经元通路 随意驾驶选择性脆弱性，并测试他们的治疗潜力。为了揭示 AD的选择性脆弱性，我们以前使用单核RNA测序提供了第一个分子 描述了人类内嗅皮层（AD早期受影响的大脑区域）中选择性脆弱的神经元 tau蛋白病理学和神经元丢失。在疾病早期丢失的神经元亚型也被选择性地 受到tau病理学的影响。这项工作为我们提供了一个相对独立的基因组之间差异表达的基因列表。 脆弱和有弹性的神经元群体。下一个挑战是确定哪些差异 基因表达的因果关系有助于选择性脆弱性。为了确定特定差异的因果作用， 表达基因的选择性脆弱性，我们将利用CRISPRi技术，使控制 内源基因的表达水平。CRISPRi由MPI博士Kampmann共同开发，他还 在人类iPSC衍生的神经元中开创了CRISPRi，并优化了其在小鼠大脑中的使用（见初步报告）。 结果）。在人类iPSC衍生神经元的全基因组CRISPRi修饰物筛选中，我们鉴定了几个 控制tau病理水平的途径。通过将CRISPRi筛选的命中与 在人类AD大脑中弹性和脆弱神经元之间的差异表达，我们发现 候选弹性因子，包括CUL 5 E3泛素连接酶复合物（Aim 1）和候选脆弱性 包括关键的糖酵解酶（Aim 2）。Aim 3的目标是进行大规模CRISPRi筛选 直接在tau蛋白病小鼠模型的脑中控制tau病理的因素。建议的重点 该项目旨在揭示AD和ADRD中选择性神经元脆弱性的潜在机制。这些 这些机制代表了潜在的治疗靶点，未来的研究将测试 针对已识别的路径。我们在这里提出的实验策略是揭示 对tau蛋白病的选择性脆弱性的潜在机制将提供一个蓝图，可以应用于许多 其他神经退行性疾病。