Mechanisms of autoimmune disease risk in IL2/IL2RA-dependent immune tolerance

IL2/IL2RA依赖性免疫耐受中自身免疫性疾病风险的机制

• 批准号: 10553203
• 负责人: Daniel J Campbell
• 金额: $ 75.42万
• 依托单位: CHILDREN'S HOSP OF PHILADELPHIA
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-02-23 至 2026-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10553203
• 关键词: 3-Dimensional; ATAC-seq; Affinity; Alleles; Animals; Antibodies; Architecture; Autoimmune; Autoimmune Diseases; Autoimmune Process; B-Lymphocytes; CD8-Positive T-Lymphocytes; CRISPR/Cas technology; Chromatin; Clinical; Complex; Cytokine Receptors; DNA; Development; Disease; Disease susceptibility; Distant; Dose; Elements; Engineering; Enhancers; Epigenetic Process; Equilibrium; Extrinsic asthma; Gene Expression; Genes; Genetic; Genetic Enhancer Element; Genetic Polymorphism; Genetic Transcription; Genetic Variation; Genomics; Graft Rejection; Homeostasis; Hormones; Human; Human Genetics; IL2 gene; IL2RA gene; Immune; Immune Tolerance; Immune response; Immunity; Infection; Inflammation; Inflammatory; Inflammatory Bowel Diseases; Insulin-Dependent Diabetes Mellitus; Intercistronic Region; Interleukin 2 Receptor; Interleukin-2; Knowledge; Link; Maps; Measures; Mediating; Modeling; Molecular; Mus; Nucleic Acid Regulatory Sequences; Population; Predisposition; Process; Recombinants; Regulator Genes; Regulatory Element; Regulatory T-Lymphocyte; Reporter; Research Institute; Role; SNP genotyping; Self Tolerance; Single Nucleotide Polymorphism; Surveys; System; Systemic Lupus Erythematosus; T cell differentiation; T cell response; T-Cell Activation; T-Lymphocyte; Testing; Untranslated RNA; Validation; Variant; autoinflammatory diseases; biobank; cytokine; design; disorder risk; epigenomics; genetic association; genetic variant; genome editing; genome wide association study; genome-wide; human disease; human subject; immune function; in vivo; insight; isoimmunity; novel; novel therapeutic intervention; organ transplant rejection; pathogen; promoter; receptor; response; risk variant; tool; trait

Project Summary Interleukin-2 is a potent T cell growth factor with crucial roles in both immunity and self-tolerance. Genome-wide association studies (GWAS) in humans have shown that genetic variation at the IL2 and IL2RA loci influence susceptibility to multiple immune-mediated diseases including allergic asthma, systemic lupus erythematosus, and inflammatory bowel disease (IBD), identifying this as a key molecular axis that controls immune activity. However, mechanistic basis of disease risk for these polymorphisms is poorly understood. We have new evidence that the cis-regulatory architectures of IL2 and IL2RA extend much further from the gene than previously appreciated, encompassing regions harboring known disease-associated variants, and we hypothesize that polymorphisms within these elements control the level and timing of IL-2 and IL-2RA expression to control the balance of tolerance vs. inflammation. In this application, we propose a comprehensive screen for potential cis-regulatory elements that interact with the IL2 and IL2RA genes using state-of-the-art epigenomic approaches like ATAC-seq and Capture-C-seq. We will establish how these elements contribute to IL2 and IL2RA gene expression using powerful, CRISPR/CAS9-based genome editing approaches. To study the impact of disease-associated genetic variation at these regulatory elements on immune function in in vivo systems, we will analyze the responses of genetically characterized subjects curated from the Benaroya Research Institute biorepository, and create mice in which orthologous murine Il2 and Il2ra enhancer sequences have been replaced with human risk alleles, assessing the disease susceptibility in models of allergic asthma, SLE and IBD. Our studies will provide comprehensive maps of the transcriptional architecture of IL2 and IL2RA, insights into the molecular basis for the genetic association of IL2 and IL2RA with autoimmune disease, and may guide the design of new approaches for the treatment of organ transplant rejection and inflammatory disease.

项目摘要 白介素-2是有效的T细胞生长因子，在免疫和自我耐受性中具有关键作用。全基因组 人类的关联研究（GWAS）表明，IL2和IL2RA基因座的遗传变异 对多种免疫介导的疾病的敏感性，包括过敏性哮喘，全身性红斑狼疮， 和炎症性肠病（IBD），将其确定为控制免疫活性的关键分子轴。 但是，这些多态性的疾病风险的机理基础知之甚少。我们有新的 IL2和IL2RA的顺式调节架构的证据比基因更远远超过了 以前被欣赏的，包括具有已知疾病相关变体的区域，我们 假设这些元素内的多态性控制IL-2和IL-2RA表达的水平和时间 控制公差与炎症的平衡。在此应用程序中，我们建议一个全面的屏幕 使用最先进的表观基因组与IL2和IL2RA基因相互作用的潜在顺式调节元素 ATAC-SEQ和CAPTURE-C-SEQ等方法。我们将确定这些要素如何对IL2和 IL2RA基因表达使用强大的，基于CRISPR/CAS9的基因组编辑方法。研究影响 这些调节元件在体内系统中免疫功能的疾病相关遗传变异的研究，我们 将分析贝纳罗瓦研究所策划的基因表征的受试者的反应 生物座，并创建了直系同源鼠IL2和IL2RA增强子序列的小鼠 取而代之的是人类风险等位基因，评估了过敏性哮喘，SLE和IBD模型中的疾病敏感性。 我们的研究将为IL2和IL2RA的转录结构提供全面的地图，对 IL2和IL2RA与自身免疫性疾病的遗传关联的分子基础，可以指导 设计器官移植排斥和炎症性疾病的新方法的设计。