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

• 批准号: 9431031
• 负责人: FRED H GAGE
• 金额: $ 161.48万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-09-30 至 2022-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9431031
• 关键词: Address; Age; Aging; Alleles; Alzheimer' s Disease; Amyloid; Anti-Inflammatory Agents; Anti-inflammatory; Appearance; Astrocytes; Atlases; Behavior; Biological Assay; Brain; Brain Diseases; CRISPR interference; Cell Line; Cell Nucleus; Cell physiology; Cells; Clustered Regularly Interspaced Short Palindromic Repeats; Coculture Techniques; Code; Data; Deposition; Disease; Disease susceptibility; Employee Strikes; Enhancers; Estrogen Receptors; Estrogens; Exhibits; Female; Fibroblasts; Gender; Gene Expression; Gene Mutation; Gene Targeting; Generations; Genes; Genetic; Genetic Enhancer Element; Genetic Transcription; Genome; Genomic approach; Guide RNA; Hippocampus (Brain); Human; Impaired cognition; Incidence; Individual; 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; 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; Address; Age; Aging; Alleles; Alzheimer' s Disease; Amyloid; Anti-Inflammatory Agents; Anti-inflammatory; Appearance; Astrocytes; Atlases; Behavior; Biological Assay; Brain; Brain Diseases; CRISPR interference; Cell Line; Cell Nucleus; Cell physiology; Cells; Clustered Regularly Interspaced Short Palindromic Repeats; Coculture Techniques; Code; Data; Deposition; Disease; Disease susceptibility; Employee Strikes; Enhancers; Estrogen Receptors; Estrogens; Exhibits; Female; Fibroblasts; Gender; Gene Expression; Gene Mutation; Gene Targeting; Generations; Genes; Genetic; Genetic Enhancer Element; Genetic Transcription; Genome; Genomic approach; Guide RNA; Hippocampus (Brain); Human; Impaired cognition; Incidence; Individual; 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; 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

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 阿尔茨海默氏病（AD）通常以特定的神经病理特征为特征，包括 细胞外淀粉样蛋白沉积物的出现和细胞内神经纤维缠结的积累。尽管 几种基因突变显然与早期发作阿尔茨海默氏病有关 表现出延迟发作的个体，与衰老相关的AD可能会带来许多链接修饰符的变化 易感性敏感性的基因。遗传和基因组广泛的关联研究（GWAS）具有 确定了许多指示细胞自主和非细胞自主的基因和风险等位基因 导致神经元丧失和认知能力下降的机制。在这方面，大多数风险变体 由GWAS鉴定出来的基因组的非编码区域，这意味着它们部分作用于改变基因 表达。该建议对RFA做出响应，表明特定的需要描述的方法 通过产生新的细胞，SNP组合的转录和细胞的后果 线试剂以帮助阐明特定神经元中的因果SNP及其靶基因的问题 源自AD个体的IPS细胞。零星广告的两个特征需要分子 解释 - 衰老在AD敏感性和引人注目的性别差异中的潜在作用，事件 广告被女性夸大。这些问题只能根据新技术和 患者衍生样品的可用性。我们提出的研究计划利用了宝贵的 在UCSD的Shiley-Marcos Alzheimer病研究中心（ADRC）的脑库中存储的样品， 以及IPSC衍生的神经元（SALK）。这种方法将质疑不同遗传变异的影响 其他危险因素（例如年龄，性别），并评估其对细胞类型特异性增强剂景观的影响。通过合并 这些数据，我们可以开始识别导致细胞类型功能改变功能的潜在的Cheusive SNP 增强剂。我们建议使用高吞吐量4C筛选方法（UMI-4C）和Hi-Chip来识别 对于功能化的编码靶基因而言，最有可能的因果关系，增强子相关的SNP。利用 当代基因编辑方法在对照或患者衍生的IPS细胞中对特定神经元细胞的功能 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 仅在孤立的细胞系中，并结合使用不同的风险等位基因组合 与星形胶质细胞和小胶质细胞进行的共培养实验，因为这些SNP的影响可能仅在 星形胶质体：神经元相互作用。这些研究将共同​​使用强大的当代全球基因组 确定增强子中几个最重要SNP的编码转录目标的方法， 以及与雌激素受体在AD性别差异中的作用的联系。