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.

项目总结 开发有效的阿尔茨海默病(AD)疾病修正疗法面临的主要挑战 和相关痴呆(ADRD)一直是我们对控制分子过程的不完全理解 发病机制。控制AD/ADRD发病的关键分子通路的重要线索可能是 从神经元对AD/ADRD选择性易感性的研究中获得。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的选择性易感性，这些通路是潜在的治疗靶点 降低神经元对疾病的易感性。因此，迫切需要发现神经通路。 随意驱动选择性易损性，并测试其治疗潜力。为了揭开决定因素 在阿尔茨海默病选择性易损性方面，我们以前使用单核RNA测序提供了第一个分子 阿尔茨海默病早期受累的人内嗅皮层选择性易损神经元的描述 由tau病理和神经元丢失引起。在疾病早期丢失的神经元亚型也是选择性的 受tau病理的影响。这项工作为我们提供了一份差异表达基因的清单， 脆弱的神经元群体与有韧性的神经元群体。下一个挑战是确定其中哪些不同 表达的基因必然会导致选择性易感性。建立特定区别的因果关系 表达基因的选择性脆弱性，我们将利用CRISPRi技术，这使得能够控制 内源基因的表达水平。CRISPRi是由MPI Dr.Kampmann共同开发的，他还 率先在人类IPSC来源的神经元中使用CRISPRi，并优化了它在小鼠大脑中的使用(见初步报告 结果)。在人类IPSC来源神经元的全基因组CRISPRi修饰物屏幕上，我们发现了几个 控制tau病理水平的途径。通过将CRISPRi屏幕上的命中结果与 我们发现，在人类AD大脑中，弹性神经元和脆弱神经元之间存在差异表达 候选复原力因素，包括CUL5 E3泛素连接酶复合体(目标1)和候选脆弱性 因素，包括关键的糖酵解酶(目标2)。目标3的目标是进行大规模CRISPRi筛查 对于tau病小鼠模型脑内直接控制tau病理的因素。建议的重点是 该项目旨在揭示AD和ADRD选择性神经元脆弱性的潜在机制。这些 机制代表了潜在的治疗靶点，未来的研究将测试 将目标锁定在已识别的路径上。我们在这里建议建立的实验策略是为了揭示 对肌萎缩侧索硬化症的选择性易感性的潜在机制将提供一张可应用于许多 其他神经退行性疾病。