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 阿尔茨海默病 (AD) 影响着全球超过 2600 万人，但治疗选择却很有限。 很大程度上是由于全基因组关联研究的分子机制尚不明确。 (GWAS) 已经确定了与 AD 相关的基因组区域，但还确定了确切的致病变异 高通量基因组技术的最新发展仍然是一个重大挑战。 在功能上表征这些因果特征具有令人难以置信的潜力，但必须应用于正确的细胞 神经元、星形胶质细胞和小胶质细胞都被认为在 AD 的病因学中发挥着关键作用，但目前尚未发现。 可以理解的是，用于这些实验的可行的原代人脑细胞是非常困难的。 快速将人类诱导多能干细胞分化为各种脑细胞类型的方案提供了巨大的帮助 有望成为破译 AD 遗传基础并指导未来治疗工作的工具。 该项目的目标是确定哪些 AD 风险变异会改变人类的转录调节能力 脑细胞并将它们映射到它们调节的基因上，以了解哪些变异在小胶质细胞中发挥作用， 星形胶质细胞和神经元，我将在人类诱导多能干细胞中进行大规模并行报告分析 细胞 (hiPSC) 分化为每种细胞类型（目标 1），为了识别 AD 风险基因，我将使用公开可用的基因。 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异与基因联系起来 （目标 2A）我将通过 CRISPR 抑制功能验证一系列假定的变异基因对（目标 2B）。 这项工作的结果将确定细胞类型特定的因果变异及其调节的基因。 将会产生积极的影响，因为准确了解哪些基因与 AD 中的哪些细胞类型有关 将阐明该疾病病因学的分子机制。本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。