Deciphering isogenic APOE isoform dependent neurodegenerative response in human glia

破译人类神经胶质细胞中同基因 APOE 亚型依赖性神经退行性反应

基本信息

批准号：
10622550
负责人：
Julia TCW
金额：
$ 12.3万
依托单位：
BOSTON UNIVERSITY MEDICAL CAMPUS
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-05-01 至 2024-02-29
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10622550
关键词：
AD transgenic mice Address Affect Age Aging Alleles Alzheimer&apos s Disease Alzheimer&apos s disease model Alzheimer&apos s disease pathology Alzheimer&apos s disease risk Amyloid beta-Protein Apolipoprotein E Astrocytes Autophagocytosis Bioinformatics Biological Assay Brain Brain region CRISPR/Cas technology Cell Differentiation process Cell physiology Cells Clinical Clinical Trials Data Development Disease Disease associated microglia E protein Environment Exhibits Future Gene Expression Gene Expression Profiling Genetic Genetic Risk Genetic Transcription Genotype Goals Human In Vitro Infant Inflammation Inflammatory Response Inherited Knock-out Knowledge Late Onset Alzheimer Disease Life Lipids Longitudinal Studies Membrane Lipids Metabolic Microglia Modality Modeling Molecular Myelin Nerve Degeneration Neurodegenerative Disorders Neurofibrillary Tangles Neuroglia Neurons Organoids Pathogenesis Pathology Pathway Analysis Patients Phagocytes Phagocytosis Phenotype Population Production Protein Isoforms Recycling Regulation Research Research Project Grants Senile Plaques Series Signal Transduction Symptoms System Systems Biology Testing Tissue-Specific Gene Expression Tissues Variant autosomal dominant Alzheimer&apos s disease brain cell cell injury cell type cytokine disorder risk embryo tissue gene network genetic risk factor genetic signature genome editing gray matter human model imaging study induced pluripotent stem cell mouse model neuroimaging neuroinflammation neuropathology novel therapeutics particle response risk variant single-cell RNA sequencing stem cell model tool transcriptome transcriptome sequencing transcriptomics

项目摘要

PROJECT SUMMARY Apolipoprotein E (APOE) is the most significant risk gene for late-onset Alzheimer's disease (AD). APOE ɛ4/ɛ4 homozygosity increases AD risk by >14-fold. Although an association between the APOE ε4 allele and increased AD risk is well-established, the mechanisms underlying this genetic risk remain elusive. In brain, microglia and astrocytes induce inflammation and degrade (engulf, digest, store or recycle) lipid-rich cellular debris that accumulates during aging and neurodegeneration. We hypothesize that APOE ɛ4/ɛ4 genotype has cell autonomous effects on astrocytes and microglia which affects other cell types. Specifically, we hypothesize that APOE ε4/ε4 astrocytes and microglia exhibit altered response to lipid-rich debris (myelin fragments), detectable as changes in global transcription and cellular function. To test this hypothesis we have generated a unique series of isogenic iPSCs differing solely in APOE isoform using a CRISPR/Cas9 genome-editing tool. We have established a platform to recapitulate cellular systems of human brain in culture by differentiation of iPSC-derived astrocytes and microglia that express APOE. We seek to bring together this established patient- derived isogenic human iPSC model, powerful systems biology, bioinformatic approaches and in vitro metabolic assays including neuroinflammation, phagocytosis and autophagy to understand the mechanism underlying APOE risk for AD. This proposed research project sets out to unravel the APOE isoform-dependent effects in glia and their responses to lipid challenge. In aim 1, we will generate homogenous populations of astrocytes and microglia and mixed cultures of cortical neurons/glia from isogenic APOE isoforms and APOE knockout derived from patient iPSCs. We will perform differential gene expression analysis to determine the downstream effects of APOE genotype in each cell type. In aim 2, we will study APOE isoform-dependent glial responses to challenge with lipid-rich particles. Following an unbiased transcriptomic approach, we will analyze which of the disease associated microglia signatures or AD-associated networks are APOE isoform-dependent upon challenge. In aim 3, we will investigate APOE isoform-dependent effects on the inflammatory response and phagocytic/autolysosomal clearance of lipid particles. The goal of this project is to identify the transcriptomic networks and cellular functions governed by APOE genotype in the presence or absence of a disease relevant environment (myelin debris) to pinpoint the earliest and potentially most treatable mechanisms involved in AD pathogenesis.

项目摘要载脂蛋白E（APOE）是晚期晚期阿尔茨海默氏病（AD）的最重要的风险基因。 apoeɛ4/ɛ4 纯合性将AD风险提高> 14倍。虽然Apoeε4等位基因与 AD风险增加是完善的，这种遗传风险的基础机制仍然难以捉摸。在大脑中，小胶质细胞和星形胶质细胞会诱导注射和降解（engulf，figest，储存或回收）富含脂质的细胞在衰老和神经变性过程中积累的碎片。我们假设APOEɛ4/ɛ4基因型具有细胞自主对星形胶质细胞和小胶质细胞影响其他细胞类型的影响。具体来说，我们假设 APOEε4/ε4星形胶质细胞和小胶质细胞暴露于对富含脂质碎屑（髓磷脂片段）的反应改变，可检测到全局转录和细胞功能的变化。为了检验这一假设，我们已经产生了仅使用CRISPR/CAS9基因组编辑工具在APOE同工型中仅在APOE同工型中区分独特的ISENIC IPSC系列。我们已经建立了一个平台，通过区分培养的人脑的细胞系统表达APOE的IPSC衍生的星形胶质细胞和小胶质细胞。我们试图将这个已建立的患者汇集在一起 - 衍生的等源性人IPSC模型，强大的系统生物学，生物信息学方法和体外代谢测定法，包括神经炎症，吞噬作用和自噬，以了解机制 APOE的潜在AD风险。该提出的研究项目旨在揭示APOE同工型依赖性神经胶质的影响及其对脂质挑战的反应。在AIM 1中，我们将产生同质种群来自等源性APOE同工型和APOE的皮质神经元/神经胶质的星形胶质细胞和小胶质细胞和混合培养物源自患者IPSC的敲除。我们将执行鉴别基因表达分析，以确定 APOE基因型在每种细胞类型中的下游效应。在AIM 2中，我们将研究依赖APOE同工型的神经胶质用富含脂质的颗粒挑战的响应。遵循公正的转录组方法，我们将分析哪些疾病相关的小胶质细胞特征或与广告相关的网络是ApoE同工型依赖性的挑战。在AIM 3中，我们将研究APOE同工型对炎症反应的影响和脂质颗粒的吞噬/自溶性清除率。该项目的目的是确定在存在或不存在的情况下，由APOE基因型控制的转录组网络和细胞功能疾病相关的环境（髓磷脂碎片），以查明最早，最可能的治疗与AD发病机理有关的机制。

项目成果

期刊论文数量（3）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Human iPSC-Based Modeling of Central Nerve System Disorders for Drug Discovery.

DOI：
10.3390/ijms22031203
发表时间：
2021-01-26
期刊：
International journal of molecular sciences
影响因子：
5.6
作者：
Qian L;Tcw J
通讯作者：
Tcw J

Human iPSC-derived astrocytes transplanted into the mouse brain undergo morphological changes in response to amyloid-β plaques.

DOI：
10.1186/s13024-021-00487-8
发表时间：
2021-09-25
期刊：
Molecular neurodegeneration
影响因子：
15.1
作者：
Preman P;Tcw J;Calafate S;Snellinx A;Alfonso-Triguero M;Corthout N;Munck S;Thal DR;Goate AM;De Strooper B;Arranz AM
通讯作者：
Arranz AM