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 晚期发病的阿尔茨海默氏病（AD），这是老年人群中最常见的痴呆形式 65是一种不可逆的，进行性脑部疾病。许多AD药物候选者旨在清除异常蛋白质 淀粉样蛋白-β（Aβ）和细胞内磷酸化TAU（P-TAU）的聚集体未能使患者受益，敦促 快速识别新的治疗靶标。 AD的全基因组关联研究（GWAS）已鉴定 越来越多的70多个风险基因座。然而，这些风险基因座的实际疾病毒害基因仍然未知。 鉴于AD的多基因病因，AD中实施了各种潜在途径。最近的进步 在基因调节网络研究中，在AD和 控制受试者已经识别并链接了功能失调的基因网络模块（或途径） 假设基因模块是分析阿尔茨海默氏病病理的靶标物质。这打开了 从网络密钥监管机构（密钥驱动程序）提名新型广告目标的新途径以调节密钥 驾驶员可能会扭转功能失调的网络，从而逆转了AD表型。网络 来自我们和其他人的分析预测了不同途径或单元类型的众多AD网络驱动程序， 但是，实验验证这些预测驱动因素的表型和基因调节作用是一个率 限制步骤。在此提案中，我们将进行基于CRISPR的屏幕，以调查功能和 扰动全基因组基因或选定的AD神经元网络面板的转录组后果 司机。我们的方法包括人类诱导的多能干细胞中的多重和精确的基因编辑 （HIPSC）使用两组CRISPR指南RNA库来衍生的神经元。通过比较grna 与AD相关的病理Aβ治疗前后细胞群中的表示，我们将确定 可以改变细胞对Aβ暴露的临界基因。平均使用单细胞转录组 多路复用细胞的测序，我们的方法将提供无偏的平行表征 每个AD神经元网络驱动基因的转录组特征。与人类一起 AD的OMICS数据，我们旨在发现AD中实施的功能失调神经元网络的新型调节剂。这 本文开发的分析和实验方法将使可扩展有效的筛选， 通过快速扩展AD中的多OMIC数据集对关键疾病网络驱动因素的测试和验证。在 长期，可以广泛应用新方法来揭示候选人的功能后果 风险基因并确定AD的新型治疗靶标。