Use of iPSC systems to define roles of microglial TREM2/DAP12 and CR3/DAP12 complexes and their genetic variants in specifying risk for late onset sporadic Alzheimer's disease

使用 iPSC 系统定义小胶质细胞 TREM2/DAP12 和 CR3/DAP12 复合物及其遗传变异在确定晚发散发性阿尔茨海默病风险中的作用

• 批准号: 10159833
• 负责人: Valentina Fossati
• 金额: $ 87.72万
• 依托单位: NEW YORK STEM CELL FOUNDATION
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-09-30 至 2023-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10159833
• 关键词: 3-Dimensional; Alleles; Alzheimer' s Disease; Alzheimer' s disease risk; Amyloid beta-Protein; Astrocytes; Autopsy; Biological; Biological Assay; Brain; CRISPR/Cas technology; Cell Culture Techniques; Cell Differentiation process; Cell Line; Cells; Clinical Pathology; Clustered Regularly Interspaced Short Palindromic Repeats; Coculture Techniques; Collaborations; Complex; Computer Analysis; Data; Data Analyses; Development; Disease; Etiology; Evaluation; Exhibits; Genes; Genetic; Genetic Polymorphism; Genetic Risk; Genetic Transcription; Genetic study; Goals; Human; Human Engineering; Image; Immune; Individual; Macrophage-1 Antigen; Maps; Microglia; Modeling; Molecular; Molecular Disease; Mus; Mutation; Neurons; Oligodendroglia; Organoids; Pathogenesis; Pathologic; Pathology; Phagocytes; Phagocytosis; Pharmaceutical Preparations; Phenotype; Population; Proteins; Protocols documentation; Reporting; Repression; Risk; Robot; Role; Series; Signal Transduction; Specific qualifier value; Structure; Synaptosomes; System; TREM2 gene; TYROBP gene; Testing; Tyrosine; Validation; Variant; Work; apolipoprotein E-4; base; brain cell; cell type; disease phenotype; disorder risk; disorder subtype; genetic manipulation; genetic variant; human data; improved; induced pluripotent stem cell; migration; molecular phenotype; molecular scale; mouse model; novel; reconstitution; relating to nervous system; response; response to injury; risk variant; screening; single cell analysis; single-cell RNA sequencing; small molecule

Project Summary Microglia are strongly implicated in the pathogenesis of Alzheimer's disease (AD) including late onset sporadic forms of the disease (LOAD). In addition to genetic studies that have identified microglial-enriched genetic variants that influence AD risk, recent computational analysis of multi-scale omics data from hundreds of human LOAD postmortem brains from our group and others in the NIA AMP-AD consortium suggest about one third of the genes associated with risk are enriched or exclusively expressed in microglia. While neurons may be the major cell type generating the toxic amyloid-beta peptide, the functional role of microglia in AD and their interaction with other cell types in the brain to cause disease are still poorly understood and the etiology of AD remains elusive. Key genetic variants in TYROBP/DAP12, TREM2, and APOE may have a functional disease altering impact in distinct brain cell type or interact across cell types in the brain. In this application, we propose to systematically identify and characterize the response of microglia to AD-associated insults in the context of these variants. To study the role of these genes and their functional interaction in AD, we will first generate a panel of CRISPR/Cas9-edited iPSC lines with isogenic mutations in TYROBP/DAP12, TREM2, and APOE in all single and multi-allelic combinations and in the context of a single genetic background with clinical and pathology confirmed LOAD. We will then generate hiPSC-derived neural co-culture systems and then complex organoids from these isogenic lines to characterize the transcriptional and functional impact of key genetic variants in single cell and cell-population-wide analyses. Single cell RNA sequencing data will be generated to identify perturbation signatures for multi-allelic variants that will then be mapped to subtype specific networks to build comprehensive signaling maps for each variant. Functional assays will be used to build evidence for relevance to AD phenotypes. Our overall goal is to test the hypothesis that genetic variants in TREM2, TYROBP/DAP12, and APOE will produce changes in iPSC-derived microglia that mimic the response of microglia to AD-associated insults.

项目总结