Mechanisms of Gut Microbiota-Driven Paneth Cell Regulation

肠道微生物驱动的潘氏细胞调节机制

• 批准号: 9973630
• 负责人: Ajay S Gulati
• 金额: $ 47.51万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-06-01 至 2025-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/9973630
• 关键词: Anti-Inflammatory Agents; Bacteria; Cations; Cell Count; Cell physiology; Cells; Cellular biology; Censuses; Clinical; Coculture Techniques; Crohn' s disease; Data; Development; Disease; Ecology; Elements; Epithelial Cells; Functional disorder; Future; Gastrointestinal Diseases; Germ-Free; Goals; Homeostasis; Housing; Human; Ice; Impairment; Individual; Inflammation; Inflammatory; Intestinal Diseases; Intestines; Knowledge; Lamina Propria; Lead; Ligands; Link; Measures; Mediating; Messenger RNA; Microbe; Mission; Modeling; Morbidity - disease rate; Mus; National Institute of Diabetes and Digestive and Kidney Diseases; Necrotizing Enterocolitis; Non-Steroidal Anti-Inflammatory Agents; Outcome; Paneth Cells; Pathogenesis; Patients; Play; Population; Probiotics; Production; Proteins; Public Health; Publishing; Regulation; Reporter; Reporting; Research; Role; Secretory Cell; Side; Signal Transduction; Small Intestines; Submucosa; Testing; Therapeutic; Therapeutic Intervention; Thromboplastin; Tissues; Work; antimicrobial; antimicrobial peptide; base; clinically relevant; design; dysbiosis; enhancing factor; enteric dysbiosis; enteric pathogen; enteritis; experimental study; graft vs host disease; gut microbiota; human disease; in vivo; innovation; insight; interleukin-22; intestinal epithelium; microbial; microbiota; mortality risk; mouse model; neutralizing antibody; new therapeutic target; novel; novel strategies; novel therapeutics; prevent; response; single-cell RNA sequencing; surgical risk; therapy development; transcriptome sequencing; treatment strategy

PROJECT SUMMARY Paneth cells (PC) are specialized small intestinal epithelial cells that control microbial populations in the gut through the secretion of antimicrobial peptides (AMPs). Importantly, PC dysfunction is emerging as a driver of disease development in several gastrointestinal (GI) disorders, including Crohn’s disease, necrotizing enterocol- itis, and graft-versus-host disease. These disorders have limited therapeutic options, which often carry significant immunosuppressive and/or surgical risks. Ultimately, developing therapies to enhance PC function may provide a novel treatment strategy for these diseases. To accomplish this, further understanding of the regulatory mech- anisms responsible for normal PC biology is required. To this end, the present study seeks to build upon previ- ously published work from this applicant’s lab, which demonstrates that the enteric microbiota significantly influ- ence PC biology. Specifically, the objective of this application is to elucidate the precise mechanisms by which the resident gut microbiota exert their regulatory effects on PCs. The clinical rationale for this work is that the knowledge obtained will support the design of targeted strategies that augment PC function to putatively treat a spectrum of GI disease. The central hypothesis of this study is that the enteric microbiota increase PC census and promote AMP production via signaling through the subepithelial tissue (i.e. tissue underlying the intestinal epithelium). This hypothesis will be tested via two independent aims that respectively focus on host and microbial elements of PC regulation. In Aim 1, we seek to determine how specific subepithelial tissue factors are able to regulate PC function. Directed by our preliminary data, we will combine (A) targeted studies focused on IL22, with (B) a broader screen of rationally-selected additional molecules (Il17a, Retnlb, Jchain, S100a9, Cxcl5) to identify novel mechanisms by which the subepithelial tissue regulates PC biology. In Aim 2, the microbial side of these interactions will be explored. This will be facilitated by a novel PC-reporter mouse (Defa6-cre;td-To- mato) we have developed, which will be studied in both conventional and germ-free housing conditions. Specif- ically, these mice will be used to: (A) determine if bacteria implicated in the pathogenesis of Crohn’s disease (which has been linked to PC dysfunction) differentially regulate PC biology; and (B) ascertain if PC function can be enhanced using the established clinical probiotic, VSL#3 – both at homeostasis and during inflammation. In combination, these aims are expected to identify host (Aim 1) and microbial (Aim 2) elements that can regulate PC function. The research proposed in this application is innovative because it explores the novel hypothesis that the subepithelial tissue is required to mediate microbial influences on PC biology. This substantially expands the conceptual framework of previous studies, which suggest that luminal bacteria directly interact with PCs to simulate antimicrobial function. The expected findings are significant because they will provide scientific justifi- cation for the development and future testing of novel agents that can clinically augment PC function. This has the potential to offer patients with limited therapeutic options new approaches for the treatment of their disease.

项目摘要 Paneth细胞（PC）是控制肠道中微生物种群的专业小肠上皮细胞 通过抗菌胡椒（AMP）的分泌。重要的是，PC功能障碍正在成为 几种胃肠道（GI）疾病的疾病发展，包括克罗恩病 ITIS和移植物与宿主病。这些疾病的治疗选择有限，通常具有重要的治疗方法 免疫抑制和/或手术风险。最终，开发增强PC功能的疗法可能会提供 这些疾病的新型治疗策略。为此，进一步了解监管机械 - 需要负责正常PC生物学的ANISM。为此，本研究旨在以预期为基础 从该申请人实验室发表的工作，这表明启动子微生物群显着影响 - ENCE PC生物学。具体而言，本应用的目的是阐明确切的机制 居民肠道微生物群对PC执行调节作用。这项工作的临床理由是 获得的知识将支持针对性策略的设计，以增强PC功能以伪造 胃肠道疾病的频谱。这项研究的中心假设是促进菌群增加了PC人口普查 并通过上皮下组织通过信号传导促进AMP的产生（即肠下属的组织 上皮）。该假设将通过两个独立目标进行检验，分别关注宿主和微生物 PC调节的要素。在AIM 1中，我们试图确定特定的下层组织因素如何能够 调节PC功能。在我们的初步数据指导下，我们将结合（a）针对IL22的目标研究， 与（b）更广泛的合理选择的额外分子（IL17A，retnlb，jchain，s100a9，cxcl5）到 确定下层组织调节PC生物学的新型机制。在AIM 2中，微生物侧 这些相互作用将被探讨。这将由一种新型的PC-Reporter小鼠（Defal6-Cre; TD-to-to-to-to-）制备 我们已经开发了Mato），这将在常​​规和无菌住房条件下进行研究。指定 - 理想情况下，这些小鼠将用于：（a）确定是否植入了克罗恩病的发病机理中 （与PC功能障碍有关）对PC生物学的调节； （b）确定PC功能是否可以 使用已建立的临床益生菌VSL＃3 - 在稳态和炎症期间都可以增强。在 组合这些目标有望识别可以调节的宿主（AIM 1）和微生物（AIM 2）元素 PC功能。该应用程序中提出的研究具有创新性，因为它探讨了新的假设 需要下层组织来介导微生物对PC生物学的影响。这大大扩展 先前研究的概念框架，这表明腔细菌直接与PC相互作用 模拟抗菌功能。预期的发现很重要，因为它们将提供科学的合理性 阳离子的开发和未来测试，可以通过临床增强PC功能的新型药物进行测试。这就是 为患者提供有限的治疗选择的潜力，用于治疗其疾病。