Intestinal Stem Cell Metabolism in Inflammatory Bowel Disease Mucosal Healing

项目摘要/摘要 炎症性肠病(IBD)是一种复杂的疾病，包括溃疡性结肠炎和克罗恩病 这会导致肠道内的长期炎症。在美国，大约有310万人 各州目前患有IBD，每年新发现7万例。这表明了一个重要的 对个人和经济造成的经济负担，在美国每年造成超过146亿美元的损失。 IBD患者经历炎症和组织损伤周期，然后是修复期和 减刑。大多数治疗IBD的免疫疗法都集中在改变免疫。 有效，但近三分之一的患者在12个月内复发。重定向我们的 关注粘膜是如何修复的可能会在IBD的治疗中提供额外的好处 免疫调节疗法。肠道干细胞(ISCs)独特的自我更新和 分化为有功能的上皮细胞使其成为组织修复不可或缺的组成部分。在 肠道，存在不同的损伤相关的ISC群体，包括在 结肠炎相关性炎症。为了更好地了解这一群体，我们开发了一种3-D老鼠器官 为这些单元格丰富的模型系统。这个系统将被用来研究细胞新陈代谢如何影响 结肠炎相关ISCs的修复功能。利用质谱仪进行代谢物分析和 使用SeaHorse XFe96分析仪的代谢生物能量分析将允许我们产生代谢 这些细胞的配置文件。我们还在开发一种新的小鼠报告线来研究葡萄糖如何 转运蛋白对结肠炎相关间充质干细胞修复功能的影响 发炎。成功完成这笔赠款将使我们能够更好地了解细胞新陈代谢是如何 结肠炎相关的ISCs影响粘膜愈合。我们的长期目标是确定结肠炎的生物标记物- 相关的ISCs可以更准确地定义粘膜愈合和疾病缓解。更好地理解 影响结肠炎相关ISC功能的分子过程将鼓励 帮助损伤的上皮细胞再生的新疗法。这项工作是在 匹兹堡大学儿童医院和匹兹堡大学仰哥研究中心， 是一个优秀的研究环境，为学员提供物质资源、研究 成功完成这项工作的队伍和智力资源。设计的详细培训计划将 帮助学员发展在ISC代谢、生物标记物发现和IBD方面的科学专长，以及 促进转变为IBD研究的独立调查者。

Project Summary/Abstract Inflammatory bowel disease (IBD), including ulcerative colitis and Crohn’s disease, is a complex disease that results in long-term inflammation within the intestine. Approximately 3.1 million individuals in the United States are currently living with IBD with 70,000 newly identified cases each year. This presents a significant financial burden on individuals and the economy, costing over $14.6 billion annually in the United States. Patients with IBD experience cycles of inflammation and tissue damage followed by periods of repair and remission. The majority of immunotherapies for IBD treatment have been focused on modifying the immune response, which are highly effective, but nearly a third of patients relapse within 12 months. Redirecting our focus on how the mucosa is repaired will likely provide an added benefit in IBD treatment when coupled with immune modulating therapies. The unique ability of intestinal stem cells (ISCs) to self-renew and differentiate into functional epithelial cells makes them an indispensable component of tissue repair. In the gut, different populations of damage-associated ISCs exist, including a ISC population that emerges during colitis-associated inflammation. To better understand this population, we developed a 3-D murine organoid model system that enriches for these cells. This system will be utilized to study how cell metabolism impacts the reparative function of colitis-associated ISCs. The use of metabolite analysis via mass spectrometry and metabolic bioenergetic analysis using a Seahorse XFe96 Analyzer will allow us to generate a metabolic profile for these cells. We are also developing a novel mouse reporter line to study how glucose transporters impact the reparative function of colitis-associated ISCs in an intestinal injury-repair model of inflammation. Successful completion of this grant will allow us to better understand how the cell metabolism of colitis-associated ISCs impacts mucosal healing. Our long-term goal is to identify biomarkers of colitis- associated ISCs to define mucosal healing and disease remission more accurately. A better understanding of the molecular processes that influence colitis-associated ISC function will encourage the development of novel therapeutics that help to regenerate the damaged epithelium. This work is being conducted at the Rangos Research Center of UPMC Children’s Hospital of Pittsburgh and the University of Pittsburgh, which is an outstanding research environment that provides the trainee with the physical resources, research cohorts, and intellectual resources to successful complete this work. The detailed training plan designed will help develop the trainee’s scientific expertise in ISC metabolism, biomarker discovery, and IBD, and facilitate the transition into an independent investigator in IBD research.