Combinatorial Actions of Genetic Variants and Gender Bias of Alzherimer's Disease

阿尔茨海默病的遗传变异和性别偏见的组合作用

基本信息

批准号：
10207441
负责人：
FRED H GAGE
金额：
$ 150.13万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN DIEGO
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-09-30 至 2023-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10207441
关键词：
Address Age Aging Alleles Alzheimer&apos s Disease Alzheimer&apos s disease brain Amyloid Anti-Inflammatory Agents Appearance Astrocytes Atlases Behavior Biological Assay Brain CRISPR interference Cell Line Cell Nucleus Cell physiology Cells Clustered Regularly Interspaced Short Palindromic Repeats Coculture Techniques Code Data Deposition Disease Disease susceptibility Enhancers Estrogen Receptors Estrogens Exhibits Female Fibroblasts Gender Gene Expression Gene Mutation Generations Genes Genetic Genetic Enhancer Element Genetic Transcription Genome Genomic approach Guide RNA Hippocampus (Brain)Human Impaired cognition Incidence Individual Induced pluripotent stem cell derived neurons Interleukin-1 Link Microglia Molecular Mutate Mutation Neurofibrillary Tangles Neurons Patients Phenotype Presenile Alzheimer Dementia Reagent Reporter Research Risk Factors Role Sampling Sex Bias Susceptibility Gene Synapses Testing Untranslated RNA Variant Vertebral column base cell type combinatorial density design epigenome excitatory neuron experimental study extracellular gender difference gender disparity genetic linkage analysis genetic variant genome wide association study genomic locus global run on sequencing induced pluripotent stem cell insight intercellular communication male mutant neuron loss new technology novel programs promoter risk variant screening sex sexual dimorphism sexual role transcriptome

项目摘要

PROJECT ABSTRACT Alzheimer’s disease (AD) is conventionally characterized by specific neuropathological features, including the appearance of extracellular amyloid deposits and the accumulation of intracellular neurofibrillary tangles. While several gene mutations are clearly associated with early onset Alzheimer’s disease, the large number of individuals exhibiting delayed onset, aging-associated AD, are likely to harbor many alterations in linked modifier genes that predispose to AD susceptibility. Genetic and genome wide association studies (GWAS) have identified numerous genes and risk alleles that indicate both cell autonomous and non-cell autonomous mechanisms contributing to loss of neurons and cognitive decline. In this regard, the majority of risk variants identified by GWAS reside in non-coding regions of the genome, implying that they act in part to alter gene expression. This proposal responds to the RFA indicating a particular need for approaches designed to delineate the transcriptional and cellular consequences of combinations of SNPs in the risk alleles by generating new cell line reagents to help unravel the question of the causative SNPs and their target genes in specific neurons derived from iPS cells of AD individuals. There are two features of sporadic AD that require molecular explanation- the potential role of aging in AD susceptibility, and the striking gender disparity, with the incidence of AD being exaggerated in females. These issues can only now be addressed based on new technologies and the availability of patient-derived samples. Our proposed research plan takes advantage of the invaluable samples stored at the brain bank of the Shiley-Marcos Alzheimer's Disease Research Center (ADRC) at UCSD, and the iPSC-derived neurons (Salk). This approach will interrogate the effects of different genetic variants with other risk factors (e.g. age, sex), and assess their effects on cell type-specific enhancer landscapes. By merging these data, we can begin to identify the potential causative SNPs that result in altered function of cell-type specific enhancers. We propose using a high throughput 4C screening approach (UMI-4C), and Hi-ChIP, to identify the most likely causative, enhancer-associated SNPs for functionally-implicated coding target genes. Exploiting the power of contemporary gene editing approaches in control or patient-derived iPS cells to specific neuronal cell types, and to astroglia, we can assess the transcriptional phenotypes and functional behaviors of neurons harboring different combinations of risk alleles, both in the isolated cell lines alone and in combination with coculture experiments with astroglia and microglia, as effects of these SNPs may be manifest only with astroglial:neuronal interactions. Together these studies will use powerful contemporary global genomic approaches to determine the coding transcriptional targets of several of the most significant SNPs in enhancers, and the link to roles of estrogen receptor in the gender disparity for AD.

项目摘要阿尔茨海默病 (AD) 通常具有特定的神经病理学特征，包括细胞外淀粉样沉积物的出现和细胞内神经原纤维缠结的积累。一些基因突变显然与早发性阿尔茨海默病有关，大量表现出延迟发作、与衰老相关的 AD 的个体可能在相关修饰因子上存在许多改变遗传和全基因组关联研究 (GWAS) 发现了易患 AD 的基因。鉴定了许多表明细胞自主和非细胞自主的基因和风险等位基因导致神经元损失和认知能力下降的机制在这方面，大多数风险变异。 GWAS 鉴定出的基因位于基因组的非编码区域，这意味着它们在一定程度上改变基因该提案是对 RFA 的回应，表明特别需要旨在描述的方法。风险等位基因中 SNP 组合通过生成新细胞产生的转录和细胞后果线试剂有助于解决特定神经元中致病 SNP 及其靶基因的问题源自 AD 个体的 iPS 细胞散发性 AD 有两个特征需要分子治疗。解释——衰老在 AD 易感性中的潜在作用，以及发病率方面显着的性别差异这些问题现在只能通过新技术和新方法来解决。我们提出的研究计划利用了宝贵的患者样本。储存在加州大学圣地亚哥分校希利-马科斯阿尔茨海默病研究中心 (ADRC) 脑库中的样本，这种方法将探究不同遗传变异的影响。其他风险因素（例如年龄、性别），并通过合并评估它们对细胞类型特异性增强子景观的影响。通过这些数据，我们可以开始识别导致细胞类型特异性功能改变的潜在致病 SNP。我们建议使用高通量 4C 筛选方法 (UMI-4C) 和 Hi-ChIP 来识别增强子。最有可能与功能相关的编码靶基因的致病性、增强子相关的 SNP。当代基因编辑方法在对照或患者来源的 iPS 细胞中对特定神经元细胞的影响类型，对于星形胶质细胞，我们可以评估神经元的转录表型和功能行为包含不同的风险等位基因组合，无论是单独的分离细胞系还是与星形胶质细胞和小胶质细胞的共培养实验，因为这些 SNP 的影响可能仅在这些研究将共同使用强大的当代全球基因组学。确定增强子中几个最重要的 SNP 的编码转录靶标的方法，以及雌激素受体在 AD 性别差异中的作用的联系。