Regulation of Intestinal Epithelial Restitution

肠上皮恢复的调节

• 批准号: 8195543
• 负责人: Jian-Ying Wang
• 金额: --
• 依托单位: BALTIMORE VA MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-04-01 至 2013-09-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8195543
• 关键词: Acute; Area; Cell Proliferation; Cell membrane; Cells; Clinic; Clinical; Complex; Coupled; Coupling; Critical Illness; DNA; Data; Disease; EF Hand Motifs; Endoplasmic Reticulum; Epithelial; Epithelial Cells; Functional disorder; Funding; Gastritis; Gastrointestinal tract structure; General Population; Genes; Goals; Health; Hemorrhage; Image Analysis; In Vitro; Incidence; Injury; Inositol; Intestines; Knowledge; Measurement; Mediating; Military Personnel; Modality; Modeling; Molecular; Mucous Membrane; Non-Steroidal Anti-Inflammatory Agents; Operative Surgical Procedures; Patients; Pattern; Pharmaceutical Preparations; Phase; Play; Population; Population Control; Prevalence; Process; Proteins; RNA Interference; RNA-Protein Interaction; Rattus; Regulation; Reporting; Role; Series; Signal Pathway; Signal Transduction; Stream; Stress; TRPC1 protein; Techniques; Testing; Therapeutic; Transfection; Ulcer; Veterans; abstracting; base; cell motility; cell type; cellular imaging; combat; design; digital; effective therapy; extracellular; gastrointestinal; human STIM1 protein; improved; in vivo; injury and repair; mutant; novel; overexpression; prevent; protein function; receptor; repaired; response; sensor; stress related disorder; trafficking; voltage

DESCRIPTION (provided by applicant): Project Summary/Abstract Early mucosal restitution is an important primary repair modality in the gastrointestinal (GI) tract and its defective regulation underlies various critical pathological states such as mucosal bleeding and acute injury, disruption of GI epithelial integrity, and barrier dysfunction. Since the exact mechanisms underlying acute mucosal injury and early rapid mucosal reepithelialization after superficial wounds are still obscure, effective therapies to preserve GI epithelial integrity in clinic are limited, especially in patients with critical surgical illnesses. During previous funding period, we have established the importance of canonical transient receptor potential-1 (TRPC1)-mediated Ca2+ signaling in regulating intestinal epithelial restitution after mucosal injury. However, the exact upstream signals initiating TRPC1 channel activation after mucosal injury remain elusive and are the focus of this competitive renewal application. Recently, stromal interaction molecule 1 (STIM1) was identified in screens for molecules that are essential for the activation of store-operated Ca2+ channels (SOCs), whereas inhibition of STIM1 expression reduces store-operated Ca2+ entry (SOCE) after store depletion. Our preliminary results indicate that a) levels of levels of STIM1 at the plasma membrane increase significantly after wounding, b) overexpression of the constitutively active STIM1 EF-hand mutant increases SOCE and enhances epithelial restitution after wounding, and c) STIM1 silencing decreases SOCE and represses epithelial restitution in cells overexpressing TRPC1. Based on these exciting observations, we HYPOTHESIZE that STIM1 plays an important role in promoting GI epithelial restitution after mucosal injury by activating TRPC1 channel activity. Three specific aims are proposed to test the hypothesis. 1) To determine the pattern and role of STIM1 in gut epithelial restitution after mucosal injury in vitro as well as in vivo. 2) To characterize functional and physical interactions of STIM1 with TRPC1 in regulating SOCE during intestinal epithelial restitution. 3) To define the cellular signaling pathways regulating STIM1 subcellular redistribution and its expression after mucosal injury. Completion of these aims will identify the up stream signals initially activating TRPC1 channels after mucosal injury and also yield a novel model in which STIM1 subcellular trafficking and expression are regulated during restitution. It is hoped that our findings will provide supportive data to strengthen our long-term goal that is to develop therapeutic approaches for GI mucosal injury-related diseases and for maintaining epithelial integrity under various clinical conditions. PUBLIC HEALTH RELEVANCE: Project Narrative Immediate goal of the current study is to define the mechanisms underlying acute gut mucosal injury and repair. Various acute GI mucosal injury and gastritis are much more common at our VA patients than those observed in the general public. It has been reported that there are 24.7% prevalence of GI ulcers and injury in veteran population, compared with 10.5% in the control population. Furthermore, drugs, specifically NSAIDs, and stress as etiologic factors in GI mucosal injury and ulcerations are becoming increasing important. This is especially relevant to the new and young OEF/OIF veterans who return with a major increase in the prevalence of stress-related disorders. The incidence of stress-induced mucosal injury and ulceration in critically ill intensive patients as well as in military combat situations further aggravate the problem of GI mucosal injury and injury-related disorders in the VA population. However, the effective therapies for preventing acute gut mucosal injury and for promoting mucosal repair in clinic are limited to date, because of lacking knowledge of the mechanisms involved in gut mucosal injury and repair. Thus, improving the understanding of the processes that maintain the integrity of the gut mucosa and enhance repair is the first step towards therapeutic initiatives in this area and is the focus of the current proposal. Based on our previous studies and exciting preliminary results, studies proposed here are to determine whether a novel protein STIM1 plays an important role in promoting GI epithelial restitution after mucosal injury by Ca2+-signaling. Completion of these aims will provide supportive data to strengthen our long-term goal that is to develop therapeutic approaches for GI mucosal injury-related diseases for our VA patients.

描述（由申请人提供）： 项目摘要/摘要早期粘膜恢复原状是胃肠道（GI）区域的重要主要修复方式及其有缺陷的调节是各种关键病理状态的基础，例如粘膜出血和急性损伤，胃肠道上皮完整性的破坏以及障碍障碍。由于浅表伤口后急性粘膜损伤和早期快速粘膜再上皮的确切机制仍然晦涩难懂，因此在临床中保留GI上皮完整性的有效疗法受到限制，尤其是在患有严重手术疾病的患者中。在以前的资金期间，我们确定了典型的瞬态受体电位1（TRPC1）介导的Ca2+信号传导在调节粘膜损伤后肠上皮恢复中的重要性。但是，粘膜损伤后启动TRPC1通道激活的确切上游信号仍然难以捉摸，并且是这种竞争性更新应用的重点。最近，在筛选中鉴定出基质相互作用分子1（STIM1）的分子，这些分子对于激活存储经营的Ca2+通道（SOCS）所必需的分子，而在存储耗竭后，STIM1表达的抑制减少了储存的CA2+进入（SOCE）。我们的初步结果表明，a）受伤后质膜上刺激的水平显着增加，b）过表达组成性主动的刺激性刺激性突变体增加了SOCE并增强了伤害后的上皮恢复，并且c）刺激症状soce soce so so so so so so s so s so s so s so s so s so s so s so cellial re nifers in Cellial在细胞中恢复过于表达trpc1。基于这些令人兴奋的观察结果，我们假设STIM1通过激活TRPC1通道活性来促进粘膜损伤后的胃肠道上皮恢复起着重要作用。提出了三个特定目标来检验该假设。 1）确定Stim1在体外和体内粘膜损伤后肠道上皮恢复的模式和作用。 2）表征STIM1与TRPC1的功能和物理相互作用，以调节肠上皮恢复过程中的SOCE。 3）定义调节STIM1亚细胞重新分布及其在粘膜损伤后表达的细胞信号通路。这些目标的完成将确定粘膜损伤后最初激活TRPC1通道的UP流信号，并产生一种新型模型，其中STIM1亚细胞运输和表达在恢复原状过程中受到调节。希望我们的发现将提供支持性数据，以增强我们的长期目标，即开发与胃肠道损伤相关疾病的治疗方法，并在各种临床状况下保持上皮完整性。 公共卫生相关性： 当前研究的项目叙述性直接目标是定义急性肠粘膜损伤和修复的基础机制。在我们的VA患者中，各种急性GI粘膜损伤和胃炎比在公众中观察到的患者更为普遍。据报道，退伍军人人口的胃肠道溃疡患病率为24.7％，而对照人群为10.5％。此外，作为胃肠道损伤和溃疡中的病因学因素，药物，特别是NSAID和应激越来越重要。这与新的和年轻的OEF/OIF退伍军人尤其重要，他们返回与压力有关的疾病的患病率的重大增加。压力诱导的粘膜损伤和严重不良的患者以及军事作战情况的发生率进一步加剧了VA人群中胃肠道粘膜损伤和与损伤有关的疾病的问题。 但是，由于缺乏了解肠粘膜损伤和修复涉及的机制，因此可以预防急性肠道粘膜损伤和促进诊所粘膜修复的有效疗法受到限制。因此，提高对维持肠粘膜完整性和增强修复过程的过程的理解是该领域治疗计划的第一步，并且是当前建议的重点。 基于我们以前的研究和令人兴奋的初步结果，这里提出的研究是确定新型蛋白质stim1在通过Ca2+信号促进粘膜损伤后促进胃肠地上皮恢复中是否起重要作用。这些目标的完成将提供支持性数据，以增强我们的长期目标，即为我们的VA患者开发与胃肠道损伤相关疾病的治疗方法。