Mechanisms of intestinal epithelial injury in intestinal inflammation

肠道炎症中肠上皮损伤的机制

• 批准号: 10533279
• 负责人: Michael George Kattah
• 金额: $ 17.21万
• 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: 2020-01-16 至 2024-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10533279
• 关键词: Acute; Adult; Affect; Antibodies; Bioinformatics; Biology; CASP8 gene; CRISPR/Cas technology; Caspase Inhibitor; Cell Death; Cell Death Induction; Cell Survival; Cell secretion; Cells; Cellular biology; Cessation of life; Clinical; Clinical Trials; Coculture Techniques; Complement; Crohn' s disease; Cytoprotection; Data; Development Plans; Disease; Epithelial Cells; Epithelium; Exhibits; Feedback; Frequencies; Genes; Genetic; Goals; Goblet Cells; Health; Health Expenditures; Hematopoietic; Hemorrhage; Histologic; Homeostasis; Hospitalization; Human; Immune; Immune system; Immunology; Immunosuppression; In Vitro; Individual; Inflammation; Inflammatory; Inflammatory Bowel Diseases; Injury; Intestines; Lead; Link; Methods; Microscopy; Modeling; Morbidity - disease rate; Mucous Membrane; Mus; Operative Surgical Procedures; Organoids; Paneth Cells; Pathogenesis; Pathogenicity; Pathway interactions; Patients; Personal Satisfaction; Pharmaceutical Preparations; Phosphorylation; Play; Predisposition; Publishing; RIPK1 gene; RNA; Refractory; Research; Research Personnel; Role; Sampling; Signal Transduction; Signaling Protein; Stimulus; Stromal Cells; System; TNF gene; Techniques; Testing; Training; Ulcerative Colitis; Viral; Work; career; career development; cell injury; colon cancer risk; disability; epithelial injury; experience; gene product; genome wide association study; gut inflammation; improved; in vivo; interest; intestinal epithelium; intestinal homeostasis; intestinal injury; kinase inhibitor; mortality; mouse model; neutralizing antibody; overexpression; preservation; prevent; risk variant; skills; small molecule; stem cell differentiation; targeted treatment

PROJECT SUMMARY AND ABSTRACT There is a great need for new treatments for Inflammatory Bowel Disease (IBD). IBD affects over 1% of US adults, leading to disability, hospitalization, and surgery. IBD is characterized by damage to the “lining” of the intestine, also known as the intestinal epithelium. Intestinal epithelial damage leads to inflammation, bleeding, and an increased risk of colon cancer. Since a damaged intestinal epithelium is the defining feature of IBD, we want to understand what genes preserve intestinal epithelial integrity in order to better treat IBD. Genome-wide association studies (GWAS) have linked hundreds of genes with small individual effects to IBD, but it is unclear how these genes cooperate to increase susceptibility to intestinal injury and inflammation. We have a mouse model to study the role of two IBD-associated genes, A20 and ABIN-1, in intestinal epithelial cells (IECs). We were the first to show that A20 and ABIN-1 work together to maintain a healthy intestinal epithelium by preventing IEC injury. We grew intestinal organoids from the intestines of these mice. Intestinal organoids are miniature replicas of the intestinal epithelium grown in a dish in the lab. Using this system, we discovered that IECs lacking A20 and ABIN-1 are exquisitely sensitive to cell death from tumor necrosis factor (TNF). TNF is one of the primary death signals in the intestine, and antibodies against TNF are commonly used to treat IBD. We have dramatic evidence that A20 and ABIN-1 cooperate to preserve intestinal well-being, but we still need to understand the specific ways in which they preserve IEC health. We will use the mouse model and our organoids to determine which inflammatory cells injure IECs in the absence of TNF. We have evidence that inflammatory cells secrete a factor or factors other than TNF that kill A20 & ABIN-1 deficient IECs. This could be a very important clue as to why some patients with IBD do not respond to anti- TNF neutralizing antibodies. Additionally, A20 and ABIN-1 might cause epithelial injury by altering how intestinal stem cells “differentiate” or mature into other IEC subsets. Two subsets of particular interest are paneth cells and goblet cells, but there are many other subsets that we will study using our unique system. Finally, and perhaps most importantly, we will also grow intestinal organoids from IBD and non-IBD patients and test susceptibility to cell death. Using the organoids, we can attempt to rescue cell death with small molecules and drugs that are currently in clinical trials. We will analyze IEC subsets in the organoids, and see how TNF and other stimuli perturb IEC subsets. Lastly, we will work on “editing” human organoids with a technique called CRISPR/Cas9, to improve our ability to study other genes in IBD. The long-term goal of these studies is to identify approaches of treating IBD that protect epithelial cells from injury, rather than simply suppressing the immune system.

项目摘要和摘要 炎症性肠病(IBD)亟需新的治疗方法。IBD影响超过1%的 美国成年人，导致残疾、住院和手术。炎症性肠病的特点是肠壁受损。 肠道，也称为肠道上皮。肠道上皮损伤会导致炎症， 出血，以及结肠癌风险的增加。因为肠道上皮受损是其定义特征 对于IBD，我们想了解哪些基因保持肠上皮完整性，以便更好地治疗IBD。 全基因组关联研究已将数百个个体效应较小的基因与IBD联系起来， 但目前尚不清楚这些基因如何协同增加对肠道损伤和炎症的易感性。 我们有一个小鼠模型来研究两个IBD相关基因A20和ABIN-1在肠道中的作用 上皮细胞(IECS)。我们是第一个证明A20和ABIN-1协同工作以保持健康的 通过预防IEC损伤来保护肠上皮细胞。我们从这些小鼠的肠道中培养出肠道有机物质。 肠道类器官是在实验室培养皿中生长的肠道上皮的微型复制品。使用这个 系统中，我们发现缺乏A20和ABIN-1的IECs对肿瘤细胞的死亡非常敏感 肿瘤坏死因子(TNF)。肿瘤坏死因子是肠道中的主要死亡信号之一，而抗肿瘤坏死因子的抗体是 通常用于治疗IBD。我们有戏剧性的证据表明A20和ABIN-1合作保护肠道 但我们仍然需要了解他们保持IEC健康的具体方式。我们将使用 小鼠模型和我们的器官，以确定哪些炎性细胞在没有肿瘤坏死因子的情况下会损伤IECS。我们 有证据表明炎症细胞分泌除肿瘤坏死因子外的一种或多种杀死A20和ABIN-1的因子 IECS缺陷。这可能是一个非常重要的线索，可以解释为什么一些IBD患者对抗炎药没有反应。 肿瘤坏死因子中和抗体。此外，A20和ABIN-1可能通过改变 肠道干细胞“分化”或成熟为其他IEC亚群。特别感兴趣的两个子集是 Paneth细胞和杯状细胞，但还有许多其他亚群，我们将使用我们独特的系统进行研究。 最后，也许最重要的是，我们还将从IBD和非IBD中培养肠道器官 并测试对细胞死亡的敏感性。使用有机化合物，我们可以尝试通过 目前正在进行临床试验的小分子和药物。我们将分析有机化合物中的IEC亚群，以及 看看肿瘤坏死因子和其他刺激是如何扰乱IEC亚群的。最后，我们将使用一个 技术称为CRISPR/Cas9，以提高我们研究IBD其他基因的能力。这些项目的长期目标是 研究是确定治疗IBD的方法，以保护上皮细胞免受损害，而不是简单地 抑制免疫系统。