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.

项目概要 晚发性阿尔茨海默病（AD）是 1 岁以上老年人群中最常见的痴呆症 65 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质。 β 淀粉样蛋白 (Aβ) 和细胞内磷酸化 tau (p-tau) 的聚集体未能使患者受益，敦促 AD 的全基因组关联研究 (GWAS) 已确定了新的治疗靶点。 越来越多的70多个风险位点，但这些风险位点的真正致病基因仍然未知。 鉴于 AD 的多基因病因学，AD 涉及多种潜在途径。 在基因调控网络研究中整合了 AD 和死后脑组织中的多组学数据 对照受试者已识别出功能失调的基因网络模块（或途径）并将其与 AD 联系起来， 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，这开启了一个新的领域。 从网络关键监管机构（关键驱动因素）提名新的 AD 目标作为调制关键的新途径 驱动程序可能会逆转功能失调的网络，从而逆转AD表型。 我们和其他人的分析预测了不同途径或细胞类型中的众多 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是一个速率 在本提案中，我们将进行基于 CRISPR 的筛选来研究其功能和功能。 扰动全基因组基因或选定的 AD 神经网络组的转录组后果 我们的方法包括对人类诱导多能干细胞进行多重和精确的基因编辑。 (hIPSC) 衍生的神经元使用两组 CRISPR 引导 RNA 文库通过比较 gRNA。 AD 相关病理性 Aβ 治疗前后细胞群中的表征，我们将确定 同时，利用单细胞转录组学研究可以改变细胞对 Aβ 暴露的反应的关键基因。 对多重扰动细胞进行测序，我们的方法将提供公正且并行的表征 每个 AD 神经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 相关的功能失调的神经网络的新调节因子。 本文开发的分析和实验方法将实现可扩展且高效的筛选， 通过 AD 中快速扩展的多组学数据集测试和验证严重疾病网络驱动因素。 从长远来看，新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 AD 的新治疗靶点。