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: 10543859
• 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/10543859
• 关键词: Actins; Address; Adult; Agonist; Animal Model; Award; Bifidobacterium; Biological Assay; Biopsy; C57BL/6 Mouse; CXCR4 Receptors; CXCR4 gene; Central Nervous System; Colitis; Collaborations; Colon; Colonoscopy; Critical Illness; Cytoskeleton; Data; Development; Distal; Enterochromaffin Cells; Enterocutaneous Fistula; Epidermal Growth Factor Receptor; Epithelial Cells; Epithelium; 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; Productivity; 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; college; commensal bacteria; epithelial repair; epithelial wound; experimental study; healing; in vivo; inhibitor; 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）在胃肠道中发挥多种生理功能， 激活14种不同的血清素受体亚型。5-羟色胺水平升高是由于组织 损伤，表明5-羟色胺释放可能介导重要的修复机制。血清素已经被证明 对皮肤损伤和胃溃疡有修复作用，5-羟色胺受体4（5-HTR 4）特别具有 已显示在结肠组织损伤后具有抗溃疡作用。然而，这背后的机制 现象尚不清楚。最近，在杯状细胞上发现了5-HTR 4，已知杯状细胞可以产生三叶草 因子3（TFF 3）;肠道修复中的关键化合物。目前，5-HTR 4和TFF 3之间的联系尚不清楚。 使用人肠类肠生成的初步数据表明，外源性5-羟色胺 刺激TFF 3释放以促进在活体成像伤口愈合测定中的修复。这一影响得到了缓解， 抑制TFF 3受体CXCR 4。在以微生物为中心的方法中，我们的实验室还发现了一种单一的 牙双歧杆菌，它在无菌小鼠以及小鼠和人类的肠中 能够刺激肠嗜铬细胞释放血清素。此外，治疗人类肠病 和B在一起在伤口愈合试验中，牙本质代谢物也增强上皮恢复，与外源性 血清素这项提议的总体假设是，血清素激活杯状细胞上的5-HTR 4， TFF 3释放，其作用于其受体CXCR 4以介导负责上皮修复的信号级联。 目的1试图使用小鼠模型确定TFF 3对5-HTR 4介导的上皮修复的体内需求 和结肠镜检查导致的结肠损伤目标2旨在研究一种微生物方法， 5-羟色胺产生和体内伤口愈合。最后，目标3解决了以下问题所需的信号级联： TFF 3介导的小鼠和人结肠肠上皮细胞的体外恢复。粘膜 愈合对于治疗溃疡、外科疾病、肠外瘘和炎性 肠道疾病伤口。未能适当愈合伤口可能导致并发症，包括感染、长时间 住院和危重病。长期目标：我的长期目标是成为一名富有成效的员工。 研究机构的独立调查员。获得K 01职业发展奖将更好地装备我 来实现这一目标。我的研究非常适合国家糖尿病、消化和肾脏研究所 与肠道伤口修复有关的疾病。我现在的机构，贝勒医学院，提供科学的 完成本提案所需的资源。另外，我召集了一批著名的科学家 我的导师委员会和微生物学和伤口修复领域的领导者担任我的导师。在 与我的导师合作，我已经制定了一个培训计划，将使我掌握先进的科学 技术，增加我的出版记录，并获得独立的资金。在这个奖项结束时，我相信 我将拥有过渡到独立的主要调查员所需的资源和专业知识。