The Role of Crypt Fissioning in IBD Ulcer Healing

隐窝分裂在 IBD 溃疡愈合中的作用

• 批准号: 10609794
• 负责人: Terrence A. Barrett
• 金额: $ 66.15万
• 依托单位: UNIVERSITY OF KENTUCKY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-02-24 至 2025-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10609794
• 关键词: Biochemical; Biogenesis; Biopsy; Childhood; Chronic; Clinical; Clinical Data; Clinical Research; Colitis; Crohn' s disease; Data; Disease; Disease Outcome; Disease remission; Drops; Epithelial Cells; Failure; Gene Expression; Generations; Genes; Genetic Transcription; Genetically Engineered Mouse; Goals; Human; In Vitro; Inflammation; Inflammatory Bowel Diseases; Injury; Intestines; Investigation; Knockout Mice; Leukocyte L1 Antigen Complex; Maintenance; Measures; Mitochondria; Modeling; Monoclonal Antibodies; Mucositis; Mucous Membrane; Mus; Operative Surgical Procedures; Outcome; Oxidation-Reduction; PTEN gene; Pathogenesis; Patients; Phosphatidylinositols; Phosphotransferases; Principal Investigator; Procedures; Refractory; Respiration; Respiratory Chain; Role; Signal Transduction; Site; TNF gene; Tattooing; Testing; Treatment Efficacy; Ulcer; Ulcerative Colitis; Visualization; WNT Signaling Pathway; Work; beta catenin; chronic ulcer; clinical phenotype; crypt cell; dextran sulfate sodium induced colitis; drug development; experience; healing; infliximab; intestinal epithelium; new therapeutic target; novel; novel strategies; prevent; programs; prospective; repaired; response; single-cell RNA sequencing; stem cells; therapeutic biomarker; therapeutic target; transcriptomics

Program Director/Principal Investigator (Barrett, Terrence, A): The importance of mucosal healing in inflammatory bowel disease (IBD) derives from clinical data that demonstrate it to be a predictor of remission1. Thus, ulcer healing predicts the capacity for therapies to prevent unwanted clinical sequelae (i.e. surgery). Unfortunately, many IBD patients remain refractory or lose responsiveness to therapies leading to persistent mucosal ulceration. Prospective pediatric studies show that reduced mitochondrial gene expression predict unfavorable outcomes in ulcerative colitis (UC)2. These findings were supported by Kugathasan et al who found that increased expression of mitochondrial respiratory chain genes predicted favorable (B1 protected) Crohn’s disease (CD) outcomes3. Based on these results and our own data, we hypothesize that a critical driver of chronic ulceration in IBD is the failure of intestinal epithelial cells (IEC) to increase mitochondrial respiration with ROS generation. We posit that chronic mucosal inflammation suppresses mitochondrial respiration needed for crypt fissioning (in ulcer healing) in IBD. This hypothesis will be tested in the following studies: Aim 1. Determine the impact of mitochondrial respiration for mucosal healing. These studies benefit from strikingly novel Prelim. Data using VilCre/mTmG/TFAMfl/fl mice where mitochondrial respiration was shown to be required for crypt fissioning in ulcer healing. In Aim 2 we will determine the requirement of mitochondrial- derived ROS in IEC crypt division using primary human colonoids. Aim 3 studies will determine the impact of chronic inflammation on ulcer healing in patients. IEC transcriptomics from ulcer edges from CD and ulcerative colitis will be compared to normal ulcer healing. Normal ulcer healing will be interrogated using a novel biopsy of a biopsy (Bx/Bx) approach (IEC will be isolated from Bx/Bx sites 4-6day after initial biopsies). ScRNAseq results have already shown that mitochondrial biogenesis (PGC1) increases in Bx/Bx ulcers whereas levels drop in IBD IEC. In summary: Studies will investigate the cause of non-healing ulceration in IBD by interrogating novel murine, human and in vitro colonoid culture models of determining the impact of suppressed mitochondrial respiration on IEC responses (crypt fissioning). The studies will test the notion that chronic mucosal inflammation (as in IBD) suppresses mitochondrial respiration which impedes ROS-induced PI3K signaling and crypt fissioning. Our goal is to identify therapeutic targets to allow for enhanced mitochondrial respiration to aide mucosal repair in IBD. OMB No. 0925-0001/0002 (Rev. 01/18 Approved Through 03/31/2020) Page Continuation Format Page

计划主任/首席研究员（Barrett，Terrence，A）： 粘膜愈合在炎症性肠病（IBD）中的重要性来自于临床数据 证明它是缓解的预测指标1。那就是溃疡治愈预测疗法的能力 不需要的临床后遗症（即手术）。不幸的是，许多IBD患者仍然难治或失去 对疗法的反应能力导致持续性粘膜溃疡。前瞻性小儿研究表明 线粒体基因表达的降低预测了溃疡性结肠炎（UC）2的不利结果。这些发现 得到了Kugathasan等人的支持，他发现线粒体呼吸链的表达增加 基因预测克罗恩病（CD）的有利（B1受保护）的结果3。基于这些结果和我们的 我们的数据，我们假设IBD中长期溃疡的关键驱动力是肠道的失败 上皮细胞（IEC）增加线粒体呼吸，随着ROS的产生。我们肯定了慢性 粘膜炎症抑制了隐窝裂变所需的线粒体呼吸（溃疡愈合） IBD。该假设将在以下研究中进行检验：目标1。确定 线粒体呼吸进行粘膜愈合。这些研究受益于新颖的预赛。 使用VILCRE/MTMG/TFAMFL/FL小鼠的数据显示，线粒体呼吸被证明是必需的 溃疡愈合中的隐窝裂变。在AIM 2中，我们将确定线粒体的要求 使用原代人类结构群在IEC Crypt分裂中得出ROS。 AIM 3研究将确定 慢性炎症对患者溃疡愈合的影响。溃疡的IEC转录组学 CD和溃疡性结肠炎的边缘将与正常的溃疡愈合进行比较。正常的溃疡愈合 将使用活检（BX/BX）方法的新型活检进行审问（IEC将与 初始活检后4-6天BX/BX站点）。 scrnaseq结果已经表明线粒体 BX/BX溃疡的生物发生（PGC1）增加，而IBD IEC的水平下降。总结：研究 将通过询问新颖的鼠类人类来研究IBD中非治疗溃疡的原因 以及确定抑制线粒体呼吸的影响的体外结肠培养模型 关于IEC响应（隐窝裂变）。研究将测试慢性粘膜的观念 炎症（如IBD中）抑制了线粒体呼吸，阻碍了ROS诱导的 PI3K信号传导和加密裂变。我们的目标是确定治疗靶标，以允许增强 IBD中辅助粘膜修复的线粒体呼吸。 OMB No. 0925-0001/0002（修订版01/18通过03/31/2020批准）页面延续格式页面