Molecular mechanisms of selective vulnerability of neurons to tauopathy

神经元选择性易损性的分子机制

基本信息

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
项目类别：
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&apos s Disease Alzheimer&apos s disease brain Alzheimer&apos s disease related dementia Alzheimer&apos 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

项目摘要

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.

项目概要开发有效的、缓解疾病的阿尔茨海默病 (AD) 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控制 AD/ADRD 发病机制的关键分子途径的重要线索可能是可以从神经元对 AD/ADRD 的选择性脆弱性的研究中获得，而不同的因素可能是不同的。导致选择性脆弱性，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，并且这些途径是 AD/ADRD 的潜在治疗靶点因此，迫切需要揭示神经元通路。随意驾驶选择性脆弱性，并测试其治疗潜力，以揭示其决定因素。 AD 中的选择性脆弱性，我们之前使用单核 RNA 测序提供了第一个分子人类内嗅皮层（AD 早期受影响的大脑区域）中选择性脆弱神经元的描述通过 tau 病理学和神经元损失，疾病早期丢失的神经元亚型也被选择性地丢失。这项工作为我们提供了相对差异表达基因的列表。下一个挑战是确定脆弱神经群体与弹性神经群体之间的差异。表达的基因对选择性脆弱性有因果作用，以建立特定差异的因果作用。在选择性脆弱性中表达基因，我们将利用 CRISPRi 技术，该技术能够控制 CRISPRi 是由 MPI Kampmann 博士共同开发的，他也是率先在人类 iPSC 衍生神经元中使用 CRISPRi，并优化了其在小鼠大脑中的使用（参见初步研究）在人类 iPSC 衍生神经元的全基因组 CRISPRi 修饰物筛选中，我们鉴定了几种。通过将 CRISPRi 筛选的命中结果与基因进行比较来控制 tau 病理学的通路水平。我们发现，人类 AD 大脑中的弹性神经元和脆弱神经元之间存在差异表达候选弹性因素，包括 CUL5 E3 泛素连接酶复合物（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 筛选。直接控制 tau 病小鼠模型大脑中 tau 病理学的因素。该项目旨在揭示 AD 和 ADRD 选择性神经脆弱性的潜在机制。机制代表了潜在的治疗靶点，未来的研究将测试其治疗潜力针对已确定的途径，我们建议建立实验策略来揭示。 tau蛋白病选择性脆弱性的潜在机制将提供一个可应用于许多领域的蓝图其他神经退行性疾病。