Defining the contribution of astrocytes to genetic MS susceptibility

确定星形胶质细胞对遗传性多发性硬化症易感性的贡献

• 批准号: 9921514
• 负责人: David Pitt
• 金额: $ 48.06万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-05-01 至 2024-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9921514
• 关键词: Activated Lymphocyte; Affect; Alzheimer' s Disease; Astrocytes; Autopsy; CRISPR/Cas technology; Cell Nucleus; Cell physiology; Cells; Chromatin; Chronic; Complex; Cytometry; Detection; Enhancers; Environment; Flow Cytometry; Freezing; Fumarates; Gene Expression Regulation; Genes; Genetic; Genetic Diseases; Genetic Transcription; Genetic study; Genotype; Histologic; Human; Image; Immune; Incentives; Individual; Infiltration; Inflammatory; Inflammatory Response; Introns; Knowledge; Lead; Lesion; Link; Lymphocyte; Maps; Mediating; Microglia; Multiple Sclerosis; Multiple Sclerosis Lesions; Myelin; Neuraxis; Neuroglia; Nuclear; Nucleic Acid Regulatory Sequences; Pathology; Pathway interactions; Peripheral; Phenotype; Predisposition; Proteins; Regulator Genes; Resources; Risk; Role; Signal Transduction; TNFRSF1A gene; Testing; Therapeutic Intervention; Tissue Banks; Tissues; Variant; base; brain cell; brain tissue; causal variant; cell type; defined contribution; epigenomics; fluorescence imaging; frontier; genetic variant; genome editing; genome wide association study; genome-wide; individualized medicine; induced pluripotent stem cell; inflammatory marker; inflammatory milieu; macrophage; multiple sclerosis patient; multiple sclerosis treatment; neurotoxicity; new technology; new therapeutic target; protective allele; recruit; response; risk variant; transcriptome; white matter

Scientific abstract Genome-wide association studies have provided >230 genetic variants associated with susceptibility to multiple sclerosis (MS). The theme emerging from these studies is that MS risk variants perturb gene regulation in activated lymphocytes, supporting the view that MS is mediated through dysregulation of lymphocytes. Absent from this picture is a role for glial cells, despite substantial evidence that these cells are critical players in MS lesion formation. In this proposal, we are challenging the lymphocyte-centric view of genetic MS susceptibility. We hypothesize that genetic variants mediate MS risk in part by dysregulating astrocyte function. Since NF-κB signaling is a central pathway in MS pathology, we will determine the impact of two common NF-κB relevant risk variants, rs1800693G (intronic to TNFRSF1A) and rs7665090G (proximal to NFKB1), on inflammatory response in astrocytes. This will be examined in human astrocyte cultures, derived from genome-edited induced pluripotent stem cells (aim 1), and in reactive astrocytes within active MS lesions (aim 2) with the risk and protective genotypes. In addition, we will quantify variant-driven changes in the lesion environment such as lymphocyte infiltration, lesion size, cellular phenotypes and phenotype interactions. In aim 3, we will systematically identify the MS risk variants that are active in astrocytes and the genes that they perturb. For this, we will generate chromatin accessibility profiles of astrocytes isolated from MS lesions that we will intersect with MS risk variants. The resulting list of variants and variant-dependent genes will provide a roadmap of astroglial pathways that are dysregulated by MS risk variants and thereby are likely to contribute to astrocyte-mediated MS susceptibility. Our results will help to define the role of astrocytes in genetically mediated MS risk. This will provide a more complete picture of how genetic variants confer susceptibility to MS. Moreover, the epigenomic profiles of astrocytes will be an important resource for research in astrocytes and can be used to investigate genetic dysregulation of astrocytes in other complex genetic disease such as Alzheimer’s disease.

科学摘要 全基因组关联研究提供了超过 230 个与多种疾病易感性相关的遗传变异 硬化症（MS）。这些研究的主题是多发性硬化症风险变异扰乱基因调控 活化的淋巴细胞，支持 MS 是通过淋巴细胞失调介导的观点。缺席的 从这张图片中可以看出神经胶质细胞的作用，尽管有大量证据表明这些细胞在多发性硬化症中发挥着关键作用 病灶形成。在本提案中，我们挑战以淋巴细胞为中心的多发性硬化症遗传易感性观点。 我们假设遗传变异部分通过星形胶质细胞功能失调介导多发性硬化症风险。由于 NF-κB 信号传导是 MS 病理学的中心途径，我们将确定两种常见 NF-κB 相关风险的影响 变体 rs1800693G（TNFRSF1A 的内含子）和 rs7665090G（NFKB1 的近端）对炎症反应的影响 在星形胶质细胞中。这将在人类星形胶质细胞培养物中进行检查，该培养物源自基因组编辑的诱导多能细胞 干细胞（目标 1），以及活动性 MS 病变内的反应性星形胶质细胞（目标 2），具有风险和保护作用 基因型。此外，我们将量化病变环境中变异驱动的变化，例如淋巴细胞 浸润、病变大小、细胞表型和表型相互作用。在目标 3 中，我们将系统地识别 在星形胶质细胞中活跃的多发性硬化症风险变异及其扰乱的基因。为此，我们将生成 从多发性硬化症病变中分离出的星形胶质细胞的染色质可及性概况，我们将其与多发性硬化症风险变异相交叉。 由此产生的变体和变体依赖性基因列表将提供星形胶质细胞通路的路线图，这些通路是 MS 风险变异失调，因此可能导致星形胶质细胞介导的 MS 易感性。 我们的结果将有助于确定星形胶质细胞在遗传介导的多发性硬化症风险中的作用。这将提供更多 全面了解遗传变异如何赋予多发性硬化症易感性。此外，表观基因组谱 星形胶质细胞将成为星形胶质细胞研究的重要资源，可用于研究遗传 其他复杂遗传疾病（如阿尔茨海默病）中星形胶质细胞的失调。