Molecular genetic analyses of transcriptional dysregulation in Alzheimers disease

阿尔茨海默病转录失调的分子遗传学分析

• 批准号: 10662322
• 负责人: Jungsu Kim
• 金额: $ 75.56万
• 依托单位: INDIANA UNIV-PURDUE UNIV AT INDIANAPOLIS
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-07-15 至 2027-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10662322
• 关键词: Address; Affect; Age; Alzheimer like pathology; Alzheimer' s Disease; Alzheimer' s disease pathology; Alzheimer' s disease patient; Alzheimer' s disease risk; Amyloid; Amyloid beta-Protein; Amyloid deposition; Amyloidosis; Animal Model; Brain; Candidate Disease Gene; Cell Line; Cell Lineage; Cells; Collaborations; Communities; Core Facility; Data; Data Set; Disease; Disease Progression; Down-Regulation; Electrophysiology (science); Frontal gyrus; Genes; Genetic Transcription; Genetic study; Gliosis; Goals; Human; Human Genetics; Immune response; Immune system; In Vitro; Inflammatory; Knock-in Mouse; Knockout Mice; Knowledge; Macrophage; Magnetic Resonance Imaging; Mediating; Metabolism; Methods; Microglia; Molecular; Molecular Genetics; Myelogenous; Neurons; Neurosciences Research; Pathogenesis; Pathology; Pathway interactions; Peptides; Phagocytes; Phagocytosis; Phenotype; Play; Positron-Emission Tomography; Proteins; Proteomics; Reporting; Role; SPI1 gene; Senile Plaques; Systems Biology; Testing; amyloid pathology; astrogliosis; chemokine; conditional knockout; cytokine; data resource; disease phenotype; epigenomics; experience; experimental study; genetic analysis; genetic risk factor; genome wide association study; genome-wide; immune function; in vitro activity; in vivo; induced pluripotent stem cell; innovative technologies; insight; knock-down; laboratory facility; lipid metabolism; monocyte; mouse model; multi-electrode arrays; multiple omics; new therapeutic target; protein aggregation; response; risk variant; selective expression; single-cell RNA sequencing; superresolution microscopy; therapy development; transcription factor; transcriptomics; uptake

Project Summary/Abstract Recently, multiple human genetic studies have identified the critical role of the immune system in the pathogenesis of Alzheimer’s disease (AD). For example, microglia phagocytose amyloid beta (Abeta) and regulate brain immune function by secreting cytokines and chemokines. Because previous studies have suggested both protective and detrimental effects of microglial activity in AD, how microglial AD risk genes, such as PU.1, affect microglial function still remains unclear. Therefore, investigating the role of AD genetic risk factors in microglia will provide critical insight into the pathobiology of the disease, potentially revealing key regulators of underlying disease mechanisms and novel therapeutic targets. An AD genetic risk factor, PU.1, is a critical transcription factor selectively expressed in microglia in the brain. We hypothesize that PU.1 affects several transcription pathways, Abeta metabolism and other AD-related pathologies by regulating their immune function. To test this hypothesis, we will apply several innovative technologies, including single-cell RNA-sequencing, quantitative proteomics, super-resolution microscopy, human-induced pluripotent stem cell-derived microglia and neurons, multi-electrode arrays, electrophysiology, and the simultaneous Positron Emission Tomography- Magnetic Resonance Imaging, in collaboration of multiple collaborators with extensive experience in these methods. In aim 1, we will determine how downregulating PU.1 affects microglial and neuronal phenotypes and perform unbiased transcriptomic and proteomic analyses to identify key downstream regulators. In aim 2, we will investigate the functional interaction between microglia and neurons using human induced pluripotent stem cells after regulating PU.1 expression. In aim 3, we will terminate how microglial PU.1 affects AD phenotypes when it is regulated before and after the onset of amyloid pathology.

项目摘要/摘要 最近，多次人类遗传研究确定了免疫系统在 阿尔茨海默氏病的发病机理（AD）。例如，小胶质细胞吞噬淀粉样蛋白β（ABETA）和 通过分泌细胞因子和趋化因子来调节脑免疫功能。因为以前的研究有 提出了小胶质细胞活性在AD中的受保护和有害作用，小胶质AD风险基因如何 作为PU.1，影响小胶质功能仍然不清楚。因此，研究AD遗传危险因素的作用 在小胶质细胞中，将为疾病的病理生物学提供关键的见解，有可能揭示关键调节剂 潜在的疾病机制和新型治疗靶标。 AD遗传危险因素PU.1是关键 转录因子在大脑的小胶质细胞中选择了转录因子。我们假设PU.1影响了几个 通过调节其免疫学功能，转录途径，ABETA代谢和其他与AD相关的病理。 为了检验这一假设，我们将采用几种创新技术，包括单细胞RNA测序， 定量蛋白质组学，超分辨率显微镜，人类诱导的多能干细胞衍生的小胶质细胞 和神经元，多电极阵列，电生理学以及同时的正电子发射断层扫描 - 磁共振成像，与多个合作者合作，拥有丰富的经验 方法。在AIM 1中，我们将确定pu.1下调如何影响小胶质细胞和神经元表型以及 执行无偏的转录组和蛋白质组学分析，以识别关键下游调节剂。在AIM 2中，我们将 使用人诱导的多能干细胞研究小胶质细胞与神经元之间的功能相互作用 调节PU.1表达后。在AIM 3中，我们将终止小胶质细胞PU.1在AD表型时如何影响AD表型 在淀粉样病理发作之前和之后受到调节。