Functional Molecular Investigation of Inflammatory Bowel Disease (IBD) Risk Variants

项目摘要/摘要 炎症性肠病是一种以间歇性为特征的慢性疾病。 肠炎发作和肠上皮屏障破坏。这个 IBD遗传学联合会已经深入研究了这种复杂疾病的遗传结构。 确定IBD风险变量的分子机制对于理解疾病至关重要 病因学和确定新的药物靶点。人类遗传学有可能提供不偏不倚的观点 疾病的致病机制。IBD已经成为密集遗传疾病的主题 调查，包括全基因组关联研究(GWAS)，发现了数十种风险 精神错乱。然而，对这些风险部位的机械性理解一直是一个挑战，因为 大多数落在基因之外，在基因组的非编码区。与之形成鲜明对比的是 在基因组的蛋白质编码区，我们了解氨基酸密码，我们仍然不知道 有一个清晰的框架来理解非编码基因组变体如何改变细胞功能和 会导致疾病。了解整个基因组中的DNA变异如何影响细胞 IBD的途径和贡献，我们现在需要更深入地了解非编码的功能 特定细胞类型中驱动病理的基因组元素。 我们提出了基于CRISPR的原代人类T细胞、人类 肠道器官(HIOS)与IBD小鼠体内病理模型的研究 IBD影响的关键顺式调控元件和功能通路 风险变种。初级免疫细胞、HIO和小鼠模型中的基因组扰动将 识别受这些关键非编码元件调控的靶基因，并识别 IBD风险变异损害基因的细胞类型或刺激特异性条件 监管。我们将评估人类所涉及的位点上的非编码变异 基因改变了静息状态下炎性和调节性T细胞的基因调节程序 状态和对刺激的反应。此外，我们还将调查以下方面的贡献 肠道增殖、上皮屏障功能障碍的特异遗传位点 使用基因编辑的完整性、自噬、细胞应激反应和再生能力 你好。了解因果非编码IBD风险变异如何扰乱人类的关键基因程序 细胞具有加速靶向治疗方法发展的潜力。

PROJECT SUMMARY/ ABSTRACT Inflammatory bowel disease (IBD) is a chronic disease characterized by intermittent episodes of intestinal inflammation and disruption of the intestinal epithelial barrier. The IBD Genetics Consortium has intensively studied the genetic architecture of this complex disease. Assigning molecular mechanisms to IBD risk variants is critical to understanding disease etiology and identify new drug targets. Human genetics has potential to provide an unbiased view of the causative disease mechanisms. IBD already has been the subject of intensive genetic investigations, including genome-wide association studies (GWAS) that have uncovered dozens of risk loci. However, mechanistic understanding of these risk loci has been a challenge because the vast majority falls outside of genes, in non-coding regions of the genome. In contrast to protein-coding regions of the genome where we understand the amino acid code, we still do not have a clear framework to understand how non-coding genome variants alter cell function and contribute to disease. To learn how DNA variation throughout the genome affects cellular pathways and contributes to IBD, we now need a deeper understanding of the function of non-coding genome elements in the specific cell types that drive the pathology. We propose targeted CRISPR-based genome perturbations in primary human T cells, human intestinal organoids (HIOs) and in vivo murine models of IBD pathology to characterize both critical cis-regulatory elements and functional pathways that are affected by IBD risk variants. Genome perturbations in primary immune cells, HIOs and murine models will identify target genes regulated by these critical non-coding elements and identify the cell-type or stimulation-specific conditions where IBD risk variants impair gene regulation. We will assess how non-coding variation at sites implicated by human genetics alters gene regulatory programs in inflammatory and regulatory T cells in both resting state and in response to stimuli. In addition, we will investigate the contribution of specific genetic loci to intestinal dysfunction in proliferation, epithelial barrier integrity, autophagy, cellular stress responses and regenerative ability using gene-edited HIOs. Understanding how causal non-coding IBD risk variants disrupt key gene programs in human cells has potential to accelerate development of targeted therapeutic approaches.