Cell-type Specific Interrogation of Variant Function in Alzheimer's Disease

阿尔茨海默病中变异功能的细胞类型特异性询问

• 批准号: 10749582
• 负责人: Marielle Louise Bond
• 金额: $ 3.91万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-08-04 至 2026-08-03
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10749582
• 关键词: ATAC-seq; Affect; Alleles; Alzheimer' s Disease; Alzheimer' s disease risk; Astrocytes; Automobile Driving; Autopsy; Binding; Biological; Biological Assay; CRISPR interference; CRISPR/Cas technology; Cell Line; Cells; Clustered Regularly Interspaced Short Palindromic Repeats; Data Set; Dementia; Development; Disease; Enhancers; Etiology; Exhibits; Future; Gene Expression Regulation; Genes; Genetic; Genetic Transcription; Genome; Genomic Segment; Genomics; Goals; Hi-C; Human; Inherited; Link; Linkage Disequilibrium; Maps; Methods; Microglia; Modeling; Molecular; Neurodegenerative Disorders; Neurons; Nucleic Acid Regulatory Sequences; Persons; Phenotype; Play; Protocols documentation; Regulatory Element; Reporter; Research; Rest; Risk; Specificity; Stimulus; Techniques; Technology; Therapeutic; Untranslated RNA; Validation; Variant; Work; brain cell; brain tissue; causal variant; cell type; design; drug development; experimental study; falls; gene product; genetic variant; genome wide association study; genomic data; induced pluripotent stem cell; multiple omics; promoter; research and development; response; risk variant; stem cell differentiation; targeted treatment; therapeutic candidate; therapeutic development; tool; transcription factor; transcriptome sequencing

Abstract Alzheimer’s Disease (AD) affects over 26 million people worldwide, yet the treatment options are limited. This is in large part due to the unclear molecular mechanisms underlying AD. Genome-wide association studies (GWAS) have identified genomic regions that are associated with AD, but determining the exact causal variants and genes remains a major challenge. Recent developments in high throughput genomic technologies have incredible potential to functionally characterize these causal features, but must be applied to the correct cell types. Neurons, astrocytes, and microglia are all thought to play key roles in the etiology of AD but obtaining viable primary human brain cells for these experiments is understandably very difficult. Therefore, recent protocols to rapidly differentiate human induced pluripotent stem cells into various brain cell types offer great promise as tools to decipher the genetic basis of AD and guide future therapeutic efforts. The goal of this project is to identify which AD risk variants alter transcriptional regulatory capacity in human brain cells and to map them to the genes they regulate. To understand which variants are functional in microglia, astrocytes, and neurons, I will perform massively parallel reporter assays in human induced pluripotent stem cells (hiPSCs) differentiated into each cell type (Aim 1). To identify AD risk genes, I will use publicly available Hi-C, ATAC-seq, and RNA-seq in hiPSC-derived microglia, astrocytes, and neurons to link variants to genes (Aim 2A). I will functionally validate a selection of putative variant-gene pairs with CRISPR inhibition (Aim 2B). The results of this work will identify cell-type specific causal variants, and the genes that they regulate. This will have a positive impact because understanding exactly which genes are involved in which cell types in AD will elucidate the molecular mechanisms of the disease etiology. The genes identified in this study can act as targets for future studies and candidates for therapeutic design.

抽象的 阿尔茨海默氏病（AD）影响了全球超过2600万人，但治疗方案却有限。这 在很大程度上是由于AD背后的分子机制不清。全基因组关联研究 （GWAS）已经确定了与AD相关的基因组区域，但确定确切的因果变异 基因仍然是一个重大挑战。高通量基因组技术的最新发展具有 在功能上表征这些因果特征的难以置信的潜力，但必须应用于正确的单元格 类型。神经元，星形胶质细胞和小胶质细胞都被认为在AD的病因中起关键作用，但获得 可以理解，这些实验的可行原代人脑细胞非常困难。因此，最近 快速将人类诱导的多能干细胞分为各种脑细胞类型的方案可提供极大的 承诺作为破译AD的遗传基础的工具，并指导未来的治疗工作。 该项目的目的是确定哪些AD风险变体会改变人类的转录调节能力 脑细胞并将其映射到它们调节的基因。要了解哪些变体在小胶质细胞中起作用， 星形胶质细胞和神经元，我将在人类诱导的多能茎中进行大量平行的记者测定 细胞（HIPSC）分化为每种细胞类型（AIM 1）。为了识别AD风险基因，我将使用公开可用 HI-C，ATAC-SEQ和RNA-SEQ在HIPSC衍生的小胶质细胞，星形胶质细胞和神经元中，将变体与基因联系起来 （AIM 2A）。我将在功能上验证具有CRISPR抑制作用的推定变体对接（AIM 2B）。 这项工作的结果将确定细胞类型的特定因果变异及其调节的基因。这 将产生积极的影响，因为确切了解哪些基因在AD中类型的细胞类型中 将阐明疾病病因的分子机制。这项研究中确定的基因可以充当 未来研究的目标和治疗设计的候选者。