Promoter interactome-aided mapping of unexplored CVID genetic landscapes

未探索的 CVID 遗传景观的启动子相互作用组辅助绘图

• 批准号: 10207385
• 负责人: NEIL David ROMBERG
• 金额: $ 57.02万
• 依托单位: CHILDREN'S HOSP OF PHILADELPHIA
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-07-16 至 2024-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10207385
• 关键词: 3-Dimensional; ATAC-seq; Affinity; Antibodies; Architecture; Autoimmune Diseases; B-Lymphocytes; BLR1 gene; Blood specimen; CD19 gene; CD4 Positive T Lymphocytes; Cataloging; Catalogs; Cell Differentiation process; Cell Line; Cells; Cessation of life; Chromatin; Coculture Techniques; Code; Combinatorics; Common Variable Immunodeficiency; Communicable Diseases; Complement 3d Receptors; Copy Number Polymorphism; DNA; Data; Data Set; Disease; Environment; Epigenetic Process; Euchromatin; Fostering; Functional disorder; Gene Expression; Genes; Genetic; Genetic Transcription; Genetic Variation; Genome; Genomic Segment; Genotype; Goals; Grant; Helper-Inducer T-Lymphocyte; Homeostasis; Hyperplasia; Immunocompetent; Immunoglobulin Class Switching; Immunoglobulin D; Individual; Inherited; Investigation; Linkage Disequilibrium; Lymph Node Dissections; Lymphocyte; Lymphocyte Function; Lymphoid Tissue; Maps; Measures; Mendelian disorder; Methods; Mind; Pathogenesis; Pathology; Patients; Phenotype; Proteins; Proxy; Publishing; Quantitative Trait Loci; RNA library; Sampling; Sentinel; Single Nucleotide Polymorphism; Spliced Genes; Structure; Structure of germinal center of lymph node; Symptoms; T-Lymphocyte; Technology; Testing; Tissues; Tonsil; Transcript; Transcriptional Regulation; Untranslated RNA; Validation; Variant; Work; cell type; chromosome conformation capture; clinically significant; common symptom; congenital immunodeficiency; exome; genetic variant; genetically modified cells; genome wide association study; genome-wide; innovation; interleukin-21; lymph node biopsy; lymph nodes; novel diagnostics; novel therapeutics; personalized medicine; programs; promoter; response; transcriptome; whole genome

PROJECT SUMMARY/ABSTRACT Common variable immunodeficiency (CVID) is a primary disease of germinal center (GC) dysfunction, yet most CVID mechanistic investigations have utilized patient blood samples, which do not contain bona fide GC cells. Similarly, genome-wide association studies (GWAS) of CVID patients have identified many disease-associated single nucleotide polymorphisms (SNPs), yet most lie in non-coding DNA and are of unclear relevance to disease pathogenesis. The long-range goal of the proposed work is to determine the full genetic basis of CVID. The objective of this grant is to determine which non-coding regions of DNA contribute to GC dysfunction. The working hypothesis is that CVID GWAS SNPs point to important non-coding genomic regions that interact with promoters of genes key to GC lymphocyte function. Further, genetic variation altering gene promoter interactions, or damage to these genes themselves, will lead to GC dysfunction and eventually the symptoms of CVID. Our rationale is that identifying gene promoter interactions critical to GC homeostasis will provide new diagnostic and therapeutic opportunities for patients suffering from diseases of GC dysfunction including, but not limited to, CVID. Our specific aims will test the following hypotheses: (Aim 1) CVID GWAS associated regions euchromatically interact with gene promoters in T follicular helper cells (Tfh) and/or follicular B cells (FO B cells); (Aim 2) these interactions influence gene expression; (Aim 3) non-coding variants altering gene expression or variants damaging coding genes are enriched in CVID patients and that rhese variants alter Tfh and FO B cell function. The contribution is significant because cataloguing the genes and gene promoter contacts vital to GC homeostasis/dysfunction will offer new disease discovery opportunities and provide new molecules to target with personalized therapies. The proposed work is innovative because we are reinterpreting published GWAS with a suite of interlocking genome-scale technologies capable of annotating individual SNPs with overlapping layers of cell type-specific epigenetic information including promoter contacts, chromatin availability and gene expression. Further, we are verifying our findings in CVID exomes and genomes, with genetically modified cell-lines, with patient lymph nodes and in co-cultures that recapitulate key transcriptional features of the GC environment.

项目总结/摘要 常见变异型免疫缺陷（CVID）是一种原发性疾病的老年中枢（GC）功能障碍，但大多数 CVID机制研究使用了不含真正GC细胞的患者血液样本。 类似地，CVID患者的全基因组关联研究（GWAS）已经确定了许多疾病相关的 单核苷酸多态性（SNPs），但大多数位于非编码DNA中，与 发病机理这项工作的长期目标是确定CVID的全部遗传基础。 该基金的目的是确定DNA的哪些非编码区导致GC功能障碍。的 工作假设是CVID GWAS SNP指向重要的非编码基因组区域， GC淋巴细胞功能关键基因的启动子。此外，改变基因启动子的遗传变异 相互作用，或对这些基因本身的损害，将导致GC功能障碍，并最终出现症状， 的CVID。我们的基本原理是，确定基因启动子相互作用的关键GC稳态将提供新的 为患有GC功能障碍疾病的患者提供诊断和治疗机会， 不限于CVID。我们的具体目标将测试以下假设：（目标1）CVID GWAS相关 与T滤泡辅助细胞（Tfh）和/或滤泡B细胞中的基因启动子相互作用的常染色区域 (FO B细胞）;（目的2）这些相互作用影响基因表达;（目的3）非编码变体改变基因表达 CVID患者中富含损伤编码基因的表达或变体，这些变体改变了Tfh 和FO B细胞功能。这一贡献是重要的，因为对基因和基因启动子进行编目 对GC稳态/功能障碍至关重要的接触将提供新的疾病发现机会， 分子来进行个性化治疗。这项提议的工作是创新的，因为我们正在重新解释 发表了GWAS，其中包含一套能够注释单个SNP的连锁基因组规模技术 具有重叠的细胞类型特异性表观遗传信息层，包括启动子接触、染色质 可用性和基因表达。此外，我们正在验证我们在CVID外显子组和基因组中的发现， 基因修饰的细胞系，与患者淋巴结和共培养物中， GC环境的特点。