Systematic identification of genetic modifiers of dysfunctional neuronal networks in Alzheimer's disease

系统鉴定阿尔茨海默病功能障碍神经元网络的遗传修饰因子

• 批准号: 10432388
• 负责人: Aiqun Li
• 金额: $ 46.48万
• 依托单位: ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-04-15 至 2025-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10432388
• 关键词: Affect; Alzheimer' s Disease; Alzheimer' s disease brain; Alzheimer' s disease pathology; Alzheimer' s disease risk; Amyloid beta-42; Amyloid beta-Protein; Automobile Driving; Autopsy; Brain; Brain Diseases; CRISPR screen; CRISPR/Cas technology; Cell Death; Cell Line; Cell model; Cells; Cellular Indexing of Transcriptomes and Epitopes by Sequencing; Clinical Trials; Clustered Regularly Interspaced Short Palindromic Repeats; Data; Data Set; Dementia; Disease; Disease Progression; Dose; Elderly; Etiology; Exposure to; Frequencies; Gene-Modified; Genes; Genetic; Genotype; Glutamates; Goals; Guide RNA; Human; Induced pluripotent stem cell derived neurons; Knock-out; Knowledge; Late Onset Alzheimer Disease; Libraries; Link; Multiomic Data; Neurons; Pathologic; Pathology; Pathway Analysis; Pathway interactions; Patients; Persons; Phenotype; Population; Population Dynamics; Proteins; Proteomics; Protocols documentation; RNA library; Regulator Genes; Research; Resolution; Role; Series; Testing; Validation; base; brain tissue; causal variant; cell type; cohort; disease phenotype; disorder control; drug candidate; gene network; gene regulatory network; genetic analysis; genome wide association study; genome-wide; human old age (65+); induced pluripotent stem cell; multiple omics; new therapeutic target; novel; novel strategies; novel therapeutics; patient stratification; protein aggregation; response; risk variant; screening; tau-1; therapeutic candidate; transcriptomics

PROJECT SUMMARY Late onset Alzheimer’s disease (AD), the most common form of dementia among elderly population of age over 65, is an irreversible, progressive brain disorder. Many AD drug candidates aiming to clear the abnormal protein aggregates of amyloid-β (Aβ) and intracellular phosphorylated tau (p-tau) have failed to benefit patients, urging a rapid identification of new therapeutic targets. Genome-wide association studies (GWAS) of AD have identified a growing number of more than 70 risk loci. Yet the real disease-causal genes at these risk loci remain unknown. Given the polygenic etiology of AD, there are various potential pathways implicated in AD. Recent advancements in gene regulatory network studies which integrate multi-Omics data in postmortem brain tissues of AD and control subjects have identified and linked dysfunctional gene network modules (or pathways) to AD, hypothesizing that gene modules are target-rich substance for analyzing Alzheimer's pathology. This opens a new avenue to nominating novel AD targets from the network key regulators (key drivers) as modulating the key drivers could potentially reverse the dysfunctional networks which in turn reverse the AD phenotypes. Network analyses from us and others have predicted numerous AD network drivers in different pathways or cell types, however, experimental validating the phenotypic and gene regulatory roles of these predicted drivers is a rate limiting step. In this proposal, we will conduct CRISPR-based screens to investigate the functional and transcriptomic consequences of perturbing genome-wide genes or a selected panel of AD neuronal network drivers. Our approach includes multiplexed and precise gene editing in human induced pluripotent stem cell (hIPSC)-derived neurons using two sets of CRISPR guide RNA libraries. By comparing the gRNA representations in the cell population before and after AD-related pathological Aβ treatment, we will identify the critical genes that can modify the cellular response to Aβ exposure. Meanwhile, using single-cell transcriptomic sequencing of the multiplex perturbed cells, our approach will provide an unbiased and parallel characterization of the transcriptomic signatures of each of the AD neuronal network driver genes. Taken together with human Omics data of AD, we aim to discover novel regulators of dysfunctional neuronal networks implicated in AD. The analytical and experimental approaches developed herein will enable the scalable and efficient screening, testing, and validation of critical disease network drivers from rapidly expanding multi-Omic datasets in AD. In the long term, the new approaches can be widely applied to reveal the functional consequences of candidate risk genes and identify novel therapeutic targets of AD.

项目摘要 晚发性阿尔茨海默病（AD）是年龄超过15岁的老年人中最常见的痴呆形式， 是一种不可逆转的进行性脑部疾病。许多AD候选药物旨在清除异常蛋白 β淀粉样蛋白（Aβ）和细胞内磷酸化tau蛋白（p-tau）的聚集未能使患者受益， 快速识别新的治疗靶点。AD的全基因组关联研究（GWAS）已经确定 70多个风险位点的数量不断增加。然而，这些风险位点上的真实的致病基因仍然未知。 鉴于AD的多基因病因学，存在与AD有关的各种潜在途径。最新进展 在整合AD死后脑组织中多组学数据的基因调控网络研究中， 对照受试者已经鉴定出功能失调的基因网络模块（或途径）并将其与AD相关联， 假设基因模块是用于分析阿尔茨海默病病理学的富含靶标的物质。这将打开一个 从网络密钥监管机构（密钥驱动程序）提名新的AD目标作为调制密钥的新途径 驱动因素可能会逆转功能失调的网络，进而逆转AD表型。网络 我们和其他人的分析已经预测了不同途径或细胞类型中的许多AD网络驱动因素， 然而，通过实验验证这些预测驱动因素的表型和基因调控作用， 限制步骤。在这项提案中，我们将进行基于CRISPR的筛选，以调查功能和 干扰全基因组基因或选定的AD神经元网络的转录组学后果 司机我们的方法包括在人类诱导多能干细胞中进行多重和精确的基因编辑， 使用两组CRISPR指导RNA文库，对hIPSC（hIPSC）衍生的神经元进行了研究。通过比较gRNA 在AD相关的病理性Aβ治疗前后，我们将确定细胞群中的表达， 这些基因可以改变细胞对Aβ暴露的反应。同时，使用单细胞转录组学 多重干扰细胞的测序，我们的方法将提供一个公正的和平行的表征 AD神经元网络驱动基因的转录组特征。与人类一起 AD的组学数据，我们的目标是发现与AD有关的功能失调的神经网络的新调节剂。的 本文开发的分析和实验方法将能够进行可扩展的和有效的筛选， 测试和验证来自AD中快速扩展的多Omic数据集的危重疾病网络驱动因素。在 从长远来看，新的方法可以广泛应用于揭示候选人的功能后果 风险基因，并确定新的治疗AD的目标。