Promoter interactome-aided mapping of unexplored CVID genetic landscapes

项目摘要/摘要 常见可变型免疫缺陷(CVID)是生发中心(GC)功能障碍的原发病，但大多数 CVID机制研究使用了患者血液样本，这些样本不包含真正的GC细胞。 同样，对CVID患者的全基因组关联研究(GWAS)已经确定了许多与疾病相关的 单核苷酸多态(SNPs)，但大多数存在于非编码DNA中，与 疾病发病机制。这项拟议工作的长期目标是确定CVID的全部遗传基础。 这项资助的目的是确定哪些DNA非编码区与GC功能障碍有关。这个 工作假说是，CVID GWASSNPs指向重要的非编码基因组区域，与 GC淋巴细胞功能关键基因的启动子。此外，基因变异改变了基因启动子 相互作用或对这些基因本身的破坏将导致GC功能障碍，并最终导致症状 是CVID的。我们的理论基础是，识别对GC动态平衡至关重要的基因启动子相互作用将提供新的 为患有GC功能障碍疾病的患者提供诊断和治疗机会，但 不限于，CVID。我们的具体目标将检验以下假设：(目标1)与CVID GWAS相关 T滤泡辅助细胞(TFH)和/或滤泡B细胞中与基因启动子等染色质相互作用的区域 (目标2)这些相互作用影响基因表达；(目标3)改变基因的非编码变体 CVID患者中破坏编码基因的表达或变体丰富，并且Rhese变体改变Tfh 和FO B细胞功能。这一贡献是显著的，因为对基因和基因启动子进行了编目 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和新的 分子以个性化治疗为靶点。拟议的工作是创新的，因为我们正在重新解释 发表了一套能够注释单个SNP的连锁基因组规模技术的GWAs 具有重叠的细胞类型特定的表观遗传信息层，包括启动子接触、染色质 可获得性和基因表达。此外，我们正在验证我们在CVID外显体和基因组中的发现， 带有患者淋巴结的转基因细胞系，并在共培养中概括关键转录 GC环境的功能。

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.