Identifying the role of serotonin receptor 4 and trefoil factor 3 in intestinal wound repair

确定血清素受体 4 和三叶因子 3 在肠道伤口修复中的作用

• 批准号: 10336138
• 负责人: Melinda Anne Engevik
• 金额: $ 12.21万
• 依托单位: MEDICAL UNIVERSITY OF SOUTH CAROLINA
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-01-13 至 2024-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10336138
• 关键词: Actins; Address; Adult; Agonist; Animal Model; Award; Bifidobacterium; Biological Assay; Biopsy; C57BL/6 Mouse; CXCR4 Receptors; CXCR4 gene; Colitis; Collaborations; Colon; Colonoscopy; Critical Illness; Cytoskeleton; Data; Development; Distal; Enterochromaffin Cells; Enterocutaneous Fistula; Epidermal Growth Factor Receptor; Epithelial; Epithelial Cells; F-Actin; Failure; Filopodia; Foundations; Funding; Future; Gastric ulcer; Gastrointestinal tract structure; Germ-Free; Gnotobiotic; Goals; Goblet Cells; HTR3A gene; Hospitalization; Human; Ileitis; Image; In Vitro; Infection; Inflammatory; Inflammatory Bowel Diseases; Institution; Intestines; K-Series Research Career Programs; Link; Literature; Mediating; Medicine; Mentors; Mentorship; Microbiology; Modeling; Molecular; Mucous Membrane; Mus; National Institute of Diabetes and Digestive and Kidney Diseases; Ondansetron; Outcome; Peripheral Nervous System; Physiological; Process; Property; Publications; Recombinants; Research; Research Personnel; Resources; Role; Scientific Advances and Accomplishments; Scientist; Secure; Serotonin; Serotonin Production; Serotonin Receptors 5-HT4; Signal Transduction; Signaling Molecule; Site; Skin; Skin injury; Surgical Anastomosis; Techniques; Testing; Therapeutic Agents; Tissues; Training; Ulcer; Work; Wound models; base; college; commensal bacteria; epithelial repair; epithelial wound; experimental study; healing; in vivo; inhibitor/antagonist; innovation; intestinal epithelium; microbial; monolayer; mouse model; novel therapeutics; prevent; receptor; release factor 3; repaired; response; sensor; serotonin receptor; tegaserod; trefoil factor; wound; wound closure; wound healing

PROJECT SUMMARY Background and aims: Serotonin (5-HT) exerts numerous physiological functions in the gastrointestinal tract via activation of 14 different serotonin receptor subtypes. Elevated serotonin levels are noted in response to tissue damage, indicating that serotonin release may mediate important repair mechanisms. Serotonin has been shown to have reparative effects on skin injury and gastric ulcers, and serotonin receptor 4 (5-HTR4) specifically has been shown to have anti-ulcerogenic action after colon tissue damage. However, the mechanism behind this phenomenon is unclear. Recently 5-HTR4 has been identified on goblet cells, which are known to produce trefoil factor 3 (TFF3); a key compound in intestinal repair. Currently the link between 5-HTR4 and TFF3 is unknown. Preliminary data generated using human intestinal enteroids has demonstrated that exogenous serotonin stimulates TFF3 release to promote repair in live imaging wound healing assays. This effect was mitigated by inhibition of the TFF3 receptor CXCR4. In a microbial-centered approach, our lab has also identified a single commensal bacterium, Bifidobacterium dentium, which in gnotobiotic mice and in mouse and human enteroids is able to stimulate serotonin release from enterochromaffin cells. Additionally, treatment of human enteroids with B. dentium metabolites also enhance epithelial restitution in a wound healing assay, similar to exogenous serotonin. The overall hypothesis of this proposal is that serotonin activates 5-HTR4 on goblet cells to stimulate TFF3 release, which acts on its receptor CXCR4 to mediate signaling cascades responsible for epithelial repair. Aim 1 seeks to define the requirement of TFF3 for 5-HTR4 mediated epithelial repair in vivo using mouse models and colonoscopy-induced colonic wounds. Aim 2 seeks to examine a microbial approach for stimulating serotonin production and wound healing in vivo. Finally, Aim 3 addresses the signaling cascade required for TFF3 mediated restitution in intestinal epithelial cells in mouse and human colonic enteroids in vitro. Mucosal healing is critical for in the treatment of ulcers, surgical anastomoses, enterocutaneous fistulae and inflammatory bowel disease wounds. Failure to properly heal wounds can result in complications including infection, prolonged hospitalization, and critical illness. Long-term objective and aims: My long-term goal is to become a productive independent investigator at research institution. Receiving a K01 Career Development Award would better equip me to achieve this goal. My research is well suited for the National Institute of Diabetes and Digestive and Kidney Diseases as it relates to intestinal wound repair. My current institution, Baylor College of Medicine, offers the scientific resources required to complete this proposal. Additionally, I have assembled a group of renowned scientists to serve on my mentorship committee and leaders in the fields of microbiology and wound repair to serve as my mentors. In collaboration with my mentors, I have developed a training plan that will allow me master advanced scientific techniques, increase my publication record and secure independent funding. At the completion of this award, I believe I will have the resources and expertise required to transition to an independent principle investigator.

项目摘要 背景和目的：5-羟色胺（5-HT）通过 14种不同的5-羟色胺受体亚型的激活。响应组织的5-羟色胺水平升高 损坏，表明5-羟色胺释放可能介导重要的修复机制。血清素已显示 对皮肤损伤和胃溃疡产生修复作用，而5-羟色胺受体4（5-HTR4）特别具有 被证明在结肠组织损伤后具有抗硫酸作用。但是，这背后的机制 现象尚不清楚。最近在杯状细胞上鉴定了5-HTR4，已知会产生Trefoil 因子3（TFF3）;肠道修复中的关键化合物。目前，5-HTR4和TFF3之间的链接尚不清楚。 使用人类肠内型产生的初步数据证明了外源5-羟色胺 刺激TFF3释放，以促进实时成像伤口愈合测定法中的修复。这种效果通过 抑制TFF3受体CXCR4。在以微生物为中心的方法中，我们的实验室还确定了一个 共生细菌，双歧杆菌牙牙，在gnotobiotic小鼠中以及小鼠和人肠内 能够刺激从肠球毒细胞中释放5-羟色胺。另外，人类肠to 与牙芽孢杆菌代谢物还增强了伤口愈合分析中的上皮恢复原状，类似于外源性 5-羟色胺。该提议的总体假设是5-羟色胺激活杯状细胞上的5-HTR4以刺激 TFF3释放，作用于其受体CXCR4以介导负责上皮修复的信号传导级联。 AIM 1试图使用鼠标模型定义TFF3对体内5-HTR4介导的上皮修复的需求 和结肠镜检查引起的结肠伤口。 AIM 2试图检查微生物方法的刺激方法 血清素的产生和体内伤口愈合。最后，AIM 3解决了所需的信号级联 TFF3在体外介导了小鼠和人类结肠肠内肠上皮细胞中的恢复。粘膜 愈合对于治疗溃疡，手术吻合，肠皮和炎症至关重要 肠道疾病伤口。无法正确治愈伤口可能会导致并发症，包括感染，长时间 住院和重症病。长期目标和目标：我的长期目标是成为一个富有成效的 研究机构的独立研究员。获得K01职业发展奖会更好地为我装备 实现这一目标。我的研究非常适合美国国家糖尿病研究所和消化研究所和肾脏 与肠道伤口修复有关的疾病。我目前的机构贝勒医学院提供了科学 完成此建议所需的资源。此外，我已经组建了一群著名的科学家来服务 在我的指导委员会以及微生物学领域和伤口修复领域的领导人中，作为我的导师。在 与我的导师合作，我制定了一项培训计划，该计划将使我掌握高级科学 技术，增加我的出版记录并获得独立资金。我相信该奖项结束后 我将拥有过渡到独立主要研究员所需的资源和专业知识。