Microbial-derived factors regulating mucosal wound healing

调节粘膜伤口愈合的微生物衍生因子

• 批准号: 10318115
• 负责人: Ruth Xinhe Wang
• 金额: $ 2.35万
• 依托单位: UNIVERSITY OF COLORADO DENVER
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-02-01 至 2022-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10318115
• 关键词: Abdominal Pain; Academic skills; Actins; Bacterial Translocation; Basic Science; Biochemical; Biomedical Research; Body Weight decreased; Butyrates; Caring; Cell Culture Techniques; Cell Shape; Cells; Chronic; Clinical; Colitis; Colonic Diseases; Diarrhea; Disease; Energy-Generating Resources; Environment; Environmental Risk Factor; Epithelial; Epithelial Cells; Equilibrium; Etiology; Fermentation; Fostering; Gastrointestinal tract structure; Genetic; Health; Hemorrhage; Histone Deacetylase; Histone Deacetylase Inhibitor; Homeostasis; Hypoxia Inducible Factor; Immune response; Immune system; Immunofluorescence Immunologic; Impaired wound healing; Inflammation; Inflammatory; Inflammatory Bowel Diseases; Injury; Intestinal Mucosa; Intestines; Link; Maintenance; Mentorship; Metabolism; Microbe; Modeling; Molecular; Morbidity - disease rate; Mucositis; Mucous Membrane; Mus; Neurons; Pathogenicity; Patients; Pattern; Persons; Physicians; Process; Production; Proteins; Quality of life; Recovery; Regulation; Relapse; Reporter; Research; Research Training; Resolution; Resources; Role; Science; Scientist; Signal Transduction; Supplementation; Surface; Testing; Therapeutic; Tight Junctions; Tissues; Training; Volatile Fatty Acids; Work; base; care costs; career; cell motility; clinical practice; colon bacteria; cytokine; epithelial wound; experience; gut inflammation; gut microbiota; host microbiome; improved; in vivo; insight; intestinal barrier; intestinal epithelium; knock-down; life time cost; loss of function; microbial; microbiota; microorganism; monolayer; murine colitis; novel; overexpression; podocyte; programs; promoter; protein expression; repaired; response; single cell sequencing; synaptopodin; synergism; tissue repair; transcription factor; translational potential; wound healing

PROJECT SUMMARY/ABSTRACT Inflammatory bowel disease (IBD) currently afflicts more than 3.1 million people in the U.S. with over 100,000 new cases each year. Patients with IBD experience persistent and relapsing gastrointestinal tract inflammation causing abdominal pain, bleeding, diarrhea, and weight loss. The etiology of IBD, while unknown, centers around the loss of intestinal barrier integrity, and comprises both genetic and environmental factors, with emerging significance of shifts in the gut microbiota. Intestinal epithelial cells (IECs) form the dynamic barrier isolating the host immune system from the external environment. Rapid wound healing after the repeated damage and barrier disruption seen in IBD is crucial to inflammatory resolution. An established role of the microbiota is production of energy in the form short-chain fatty acids (SCFAs), such as butyrate. Decreases in butyrate-producing species are strongly associated with IBD. Preliminary studies show that butyrate augments barrier formation and enhances epithelial wound healing following injury. An unbiased single cell sequencing screen revealed that butyrate induces IEC expression of synaptopodin (SYNPO), an actin-associated protein previously uncharacterized in the intestinal epithelium. This proposal will test the hypothesis that the microbial-derived SCFA butyrate promotes intestinal wound healing and barrier through coordination of SYNPO expression and function in the context of inflammation resolution as well as homeostatic maintenance. Three specific aims will guide this project. Aim 1 will define the mechanisms of SYNPO regulation by SCFAs, including butyrate, through cell culture and promoter reporter analysis. Aim 2 will elucidate the functional role of SYNPO in IECs utilizing knockdown and overexpression cells and immunofluorescence. Aim 3 will determine the contribution of SYNPO in health and during mucosal disease using murine colitis models. Successful completion of this work will establish a critical role for the microbiota in regulating wound healing and ultimately recovery from IBD through a novel target, SYNPO. Understanding the mechanisms through which butyrate repairs tissue damage and restores the intestinal barrier will contribute to current therapeutic approaches. This comprehensive research training plan will provide outstanding mentorship with an experienced sponsor in the ideal environment of a rigorous basic science lab that is well-integrated clinically with the necessary resources for completing each aspect of this project. This includes a distinct mentorship team within the Mucosal Inflammation Program in addition to the guidance of the applicant’s thesis committee. This training will foster the applicant’s research and academic skills to pursue cross-cutting molecular level science that will advance therapeutics for tissue damage repair and novel disease target identification. These mechanistic studies hold translational potential to improve the quality of life for IBD patients and provide the optimal progression towards a career balancing biomedical research and clinical practice as a physician scientist.

项目概要/摘要 目前，美国有超过 310 万人患有炎症性肠病 (IBD)，其中超过 10 万人患有炎症性肠病 (IBD) 每年都有新病例出现持续性和复发性胃肠道炎症。 引起腹痛、出血、腹泻和体重减轻 IBD 的病因虽然未知，但主要集中在以下方面： 肠道屏障完整性的丧失，包括遗传和环境因素， 肠道微生物群变化的重要性肠上皮细胞（IEC）形成隔离肠道菌群的动态屏障。 宿主免疫系统免受外界环境的反复损伤和屏障后伤口快速愈合。 IBD 中的破坏对于炎症消退至关重要。 短链脂肪酸 (SCFA) 形式的能量，例如丁酸 产生丁酸的物质减少。 初步研究表明丁酸盐可增强屏障形成并与 IBD 密切相关。 增强损伤后的上皮伤口愈合 无偏见的单细胞测序筛选表明 丁酸盐诱导突触蛋白 (SYNPO) 的 IEC 表达，突触蛋白是一种肌动蛋白相关蛋白 该提案将检验微生物来源的假设。 SCFA 丁酸盐通过协调 SYNPO 表达和促进肠道伤口愈合和屏障 在炎症消退和稳态维持方面的功能将实现三个具体目标。 目标 1 将通过 SCFA（包括丁酸盐）定义 SYNPO 监管机制。 目标 2 将利用细胞培养和启动子报告基因分析阐明 SYNPO 在 IEC 中的功能作用。 敲低和过表达细胞以及免疫荧光将确定 SYNPO 的贡献。 使用小鼠结肠炎模型在健康和粘膜疾病期间的成功完成这项工作将。 通过以下方式确定微生物群在调节伤口愈合和最终 IBD 恢复中的关键作用： 一个新的目标，SYNPO 了解丁酸修复组织损伤的机制 恢复肠道屏障将有助于当前的治疗方法。 这项全面的研究培训计划将由经验丰富的赞助商提供出色的指导 在严格的基础科学实验室的理想环境中，该实验室与临床和必要的良好集成 完成该项目各个方面的资源，其中包括粘膜内部的独特指导团队。 除了申请人论文委员会的指导外，该培训还将促进炎症计划。 申请人的研究和学术技能，以追求跨领域的分子水平科学，这将推动 这些机制研究证明了组织损伤修复和新疾病靶点识别的治疗方法。 改善 IBD 患者生活质量并提供最佳进展的转化潜力 作为一名医师科学家，平衡生物医学研究和临床实践的职业。