Using human intestinal organoids to model IBD pathogenesis

使用人类肠道类器官模拟 IBD 发病机制

• 批准号: 10264945
• 负责人: Ophir D Klein
• 金额: $ 24.23万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-16 至 2022-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10264945
• 关键词: Autophagocytosis; Bacteria; Biochemical; Biological; Biological Markers; Biology; CRISPR/Cas technology; Cell physiology; Cells; Cellular Stress; Chronic Disease; Collaborations; Complex; Couples; Data; Developed Countries; Development; Diagnostic tests; Disease; Disease Progression; Ensure; Environmental Risk Factor; Epigenetic Process; Epithelial; Etiology; Foundations; Functional disorder; Future; Gene Expression Regulation; Genes; Genetic; Genetic Diseases; Genetic Variation; Genetic study; Genomics; Goals; Human; Human Genetics; Immunologic Surveillance; Incidence; Individual; Inflammatory Bowel Diseases; Intestines; Investigation; Learning; Link; Modeling; Molecular; National Institute of Diabetes and Digestive and Kidney Diseases; Organoids; Pathogenesis; Pathogenicity; Pathology; Pathway interactions; Patients; Phenotype; Prevalence; Regulation; Research; Risk; Role; Susceptibility Gene; Testing; Tissues; adaptive immune response; base; causal variant; cell type; disorder control; disorder risk; drug discovery; genetic architecture; genetic manipulation; genome wide association study; genomic locus; human embryonic stem cell; human population study; inflammatory disease of the intestine; insight; interest; intestinal crypt; intestinal epithelium; intestinal homeostasis; mutant; new therapeutic target; novel diagnostics; programs; regeneration following injury; regenerative; response; risk variant; targeted treatment; tissue regeneration; treatment response; trend

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 for identification of new drug targets. Human genetics has the 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, and in this exploratory R21 proposal, we will employ two functional models – patient-derived enteroids and human intestinal organoids (HIOs) – to study the role of several important genes in epithelial restitution during IBD. This project depends on close collaboration with the NIDDK IBD Genetics Consortium (IBDGC) for two reasons. First, we will benefit from early access to IBDGC data to ensure that we deploy our pipeline for functional characterization of the highest priority IBD risk variants. Second, we will access patient-derived enteroids as a collaboration with the IBDGC. Our central goal is functional characterization of the biological pathways disrupted by IBD risk variants, which we will accomplish with two specific aims. In Aim 1, we will use gene editing to delete genetic loci, produce HIOs, and study developmental role of genes of interest. In Aim 2, we will investigate the contribution of specific genetic loci to intestinal dysfunction in proliferation, epithelial barrier integrity, autophagy, cellular stress, and regenerative ability using patient-derived enteroids and gene-edited HIOs.

摘要 炎症性肠病是一种以肠道间歇性发作为特征的慢性疾病。 肠道上皮屏障的炎症和破坏。IBD遗传学联盟密集地 研究了这种复杂疾病的遗传结构。IBD风险变异的分子机制研究 对于理解疾病病因学和确定新药靶点至关重要。人类遗传学具有 有可能为致病机制提供一个公正的观点。IBD已经成为主题 密集的遗传研究，包括全基因组关联研究(GWAS) 几十个风险基因。然而，对这些风险部位的机械性理解一直是一个挑战，在这方面 探索性的R21提案，我们将使用两种功能模型-患者来源的肠样和人类肠道 有机物(HIO)-研究几个重要基因在IBD期间上皮重建中的作用。这个项目 依赖于与NIDDK IBD遗传学联合会(IBDGC)的密切合作有两个原因。首先，我们 将受益于早期访问IBDGC数据，以确保我们部署我们的功能表征流水线 最高优先级的IBD风险变种。其次，我们将与患者合作访问患者来源的肠样肉 IBDGC。我们的中心目标是对IBD风险扰乱的生物途径进行功能表征 变种，我们将通过两个具体目标来实现。在目标1中，我们将使用基因编辑来删除基因 基因座，产生HIO，并研究感兴趣基因的发育作用。在目标2中，我们将调查 特定基因座对肠功能障碍在增殖、上皮屏障完整性、自噬、 细胞应激，以及使用患者来源的肠样组织和基因编辑的HIO的再生能力。