Prioritizing and Characterizing T Cell-Relevant Genetic Variants Associated with Autoimmune Diseases

优先考虑和表征与自身免疫性疾病相关的 T 细胞相关遗传变异

• 批准号: 10320334
• 负责人: John Philip Ray
• 金额: $ 10.8万
• 依托单位: BENAROYA RESEARCH INST AT VIRGINIA MASON
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-12-18 至 2022-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10320334
• 关键词: Address; Affect; Alleles; Architecture; Autoimmune Diseases; Autoimmunity; BACH2 gene; Biological Assay; CRISPR interference; Cell Line; Cell physiology; Cells; ChIP-seq; Chromatin; Clustered Regularly Interspaced Short Palindromic Repeats; Complex; Disease; Disease susceptibility; Elements; Engineering; Epigenetic Process; Fluorescence-Activated Cell Sorting; Follow-Up Studies; Gene Expression; Genes; Genetic; Genetic Determinism; Genetic Variation; Genome; IRF1 gene; Immune; Immunohistochemistry; Inflammatory Bowel Diseases; Insulin-Dependent Diabetes Mellitus; Linkage Disequilibrium; Mediating; Multiple Sclerosis; Nucleic Acid Regulatory Sequences; Pathogenesis; Pathogenicity; Pathway interactions; Patients; Population; Psoriasis; Psoriatic Arthritis; Reading; Regulatory Element; Reporter; Rest; Rheumatoid Arthritis; STAT4 gene; T-Cell Activation; T-Lymphocyte; Testing; Therapeutic; United States; Untranslated RNA; Validation; Variant; Work; base; base editing; causal variant; cell type; exhaustion; follow-up; genetic variant; genome resource; genome wide association study; genomic data; insight; mRNA Expression; new therapeutic target; novel therapeutics; protein expression; response; transcription factor

PROJECT SUMMARY Greater than 8% of the United States population suffers from autoimmune disease, but, due to complex non- Mendelian inheritance, the genetic determinants of autoimmune disease are difficult to parse. To begin to address this problem, genome-wide association studies (GWAS) have identified thousands of genetic variants that track with disease, allowing the field of autoimmunity to focus on key disease-causal regions of the genome. However, the exact causal genetic variants for most of these associations remain unidentified, and thus the genes and pathways they alter remain poorly understood. To tackle this problem, I first will enrich for likely causal variants for diseases in which T cells are known to be pathogenic, including multiple sclerosis, type I diabetes, rheumatoid arthritis, psoriasis, and inflammatory bowel disease. I will use a high-throughput approach to test 20,000 variants for allelic skew in reporter expression. Furthermore, for 4 highly important GWAS loci (each with more than 10 disease associations), I will screen for regulatory regions that alter gene expression and the disease-associated variants that lie within these regions. With these two approaches (and other genomic data, such as chromatin accessibility and allele-specific transcription factor ChIP-seq), I will prioritize variants for engineering in the genomes of primary cells, and determine the effects of these engineered alleles on expression, activation, and polarization of T cells. This project will result in exhaustive characterization of variants associated to 5 important autoimmune diseases, elucidation of the regulatory architecture of 4 highly important disease loci, and experimental validation of 10 putatively causal variants through editing them into the genome of primary T cells. This work will provide an extensive resource for GWAS follow-up studies, help bring the field closer to understanding the pathways and regulatory architecture involved in disease, and inform approaches for identifying new targeted therapeutics for autoimmunity.

项目摘要 超过8%的美国人口患有自身免疫性疾病，但是，由于复杂的非免疫性疾病， 孟德尔遗传，自身免疫性疾病的遗传决定因素很难解析。开始 为了解决这个问题，全基因组关联研究（GWAS）已经确定了数千种遗传变异 跟踪疾病，使自身免疫领域专注于关键的疾病因果区域， 基因组然而，大多数这些关联的确切因果遗传变异仍然没有确定， 因此，它们改变的基因和途径仍然知之甚少。为了解决这个问题，我首先将丰富 已知T细胞是致病性的疾病的可能致病变体，包括多发性硬化症， I型糖尿病、类风湿性关节炎、牛皮癣和炎症性肠病。我会用高通量的 方法来测试20，000个变体在报告基因表达中的等位基因偏斜。此外，对于4个非常重要的 GWAS基因座（每个基因座与10种以上的疾病相关），我将筛选改变基因的调控区域， 表达和位于这些区域内的疾病相关变体。通过这两种方法（以及 其他基因组数据，如染色质可及性和等位基因特异性转录因子ChIP-seq），我将 优先考虑原代细胞基因组中的工程变体，并确定这些变体的影响。 在一些实施方案中，本发明的抗体可以用于治疗工程化等位基因对T细胞的表达、活化和极化的影响。该项目将导致详尽的 表征与5种重要自身免疫性疾病相关的变体，阐明调节免疫应答的机制。 4个高度重要的疾病基因座的结构，以及10个pupillary致病变异体的实验验证 通过将它们编辑到原代T细胞的基因组中。这项工作将提供广泛的资源， GWAS的后续研究，有助于使该领域更接近于了解途径和监管架构 参与疾病，并为识别新的自身免疫靶向治疗方法提供信息。