Targeting Intestinal Epithelium to Promote Healing in Inflammatory Bowel Disease

项目总结/摘要 方法： 我将重点介绍NR 5A核受体肝脏受体同源物-1（LRH-1，NR 5A 2），越来越多的人认识到， 因为它在细胞生长和抗炎转录中的作用。缺乏肠道LRH-1敲除的小鼠 显示IBD模型中粘膜损伤和炎症标志物增加。我假设LRH-1的缺失 将损害上皮更新和对损伤的反应，并且增强的LRH-1表达将抑制 肠道炎症和加速粘膜愈合。我开发了一种离体肠道类器官培养方法 （肠）系统以研究上皮对损伤的反应。肠道系统让我可以直接解决 LRH-1在肠上皮病理生理学中的作用，通过使用条件性小鼠遗传模型， 肠上皮中的LRH-1（Villin-CreER）。我已经调整了腺相关病毒（AAV）系统， 能够表达人LRH-1和肠样系统中重要的LRH-1突变， LRH-1介导的上皮功能。我将使用具有条件性LRH-1敲除和条件性LRH-1敲除的体内IBD模型。 过度表达，以建立我的体外研究，并确定LRH-1在上皮损伤和 炎症反应。我的工作将确定LRH-1的激活是否足以减弱免疫介导的 粘膜损伤，并询问LRH-1是否可以用于IBD药物开发。我相信这些问题 在这个基于Mentored的K 08应用程序中概述的实验模型是令人兴奋的， 与我对炎症性肠道疾病的临床兴趣有关。 候选人和理由： 我是一个双学位医师科学家MSTP研究生在胃肠病学和药理学培训。在这 我试图了解控制肠道内稳态的分子途径和适应性的 驱动损伤反应的机制。在肠道中，炎症反应由两种途径介导。 代谢和环境线索。炎症性肠病（IBD）是一种慢性疾病， 成人和儿童都一样，造成显著的发病率和医疗保健成本。了解如何 肠上皮自我更新，并保持其完整性，面对炎症损伤的关键是， 开发用于肠道疾病的新疗法。我的最终目标是领导一项独立的研究 该计划侧重于翻译肠道疾病模型和治疗设计。 职业目标： 通过这个建议所划定的框架，我将建立一个独立的研究计划 致力于了解肠道病理生理学和新疗法的发展。的 本文描述的研究计划旨在将我的专业知识扩展到动物遗传学的新领域， 疾病建模这与我的转化科学的长期目标是一致的， 让我的实验室造福于我的病人我将得到我的咨询委员会的指导， 在加州大学旧金山分校的职业发展研讨会，如赠款和手稿写作，将培养我的两个 专业和科学发展。 工作环境： 我已经确定了一个强大的指导团队，由核受体生物学的领导者冬青博士英格拉哈姆领导。博士 Averil Ma是IBD和结肠炎动物模型的专家，Ophir Klein博士是内科科学家， 上皮干细胞专业知识，完善我的顾问委员会与互补的技能组合。来增强我 研究培训，我将从事教学课程在免疫学，先进的光学显微镜，高 吞吐量数据分析和负责任的研究行为。UCSF提供了一个刺激的环境 为了追求科学与高度合作的调查网络。儿科系和 胃肠病学分部在基础和临床方面都有培养未来领导者的悠久历史 科学 总之，作为一个积极主动的临床科学家，我的最终目标是开发一个独立的基础和 翻译研究计划，重点是肠道疾病。该建议提供了必要的培训， 研究加上强大的指导计划，以帮助我取得成功。

PROJECT SUMMARY/ABSTRACT Approach: I will focus on the NR5A nuclear receptor Liver Receptor Homolog-1 (LRH-1, NR5A2), increasingly recognized for its roles in cell growth and its anti-inflammatory transcription profile. Mice lacking intestinal LRH-1 knockout show increased mucosal damage and inflammatory markers in IBD models. I hypothesize that loss of LRH-1 will impair epithelial renewal and response to injury, and that enhanced LRH-1 expression will dampen intestinal inflammation and speed mucosal healing. I have developed an ex vivo intestinal organoid culture (enteroid) system to study epithelial responses to injury. The enteroid system allows me to directly address the role of LRH-1 in intestinal epithelial pathophysiology by using conditional mouse genetic models that eliminates LRH-1 in the intestinal epithelium (Villin-CreER). I have adapted the adeno-associated virus (AAV) system to enable expression of human LRH-1 and important LRH-1 mutations in the enteroid system to further probe LRH-1-mediated epithelial function. I will use in vivo IBD models with both conditional LRH-1 knockout and overexpression to build upon my ex vivo studies, and to define the role of LRH-1 in epithelial injury and inflammatory response. My work will determine if LRH-1 activation is sufficient to blunt immune-mediated mucosal injury and ask if LRH-1 can be targeted for IBD drug development. I believe that the questions being asked and the experimental models outlined in this Mentored-based K08 application are exciting and highly relevant to my clinical interests in inflammatory intestinal disorders. Candidate & Rationale: I am a dual-degree physician scientist MSTP graduate trained in gastroenterology and pharmacology. In this proposal I seek to understand the molecular pathways that govern bowel homeostasis and the adaptive mechanisms that drive injury response. In the intestine, inflammatory responses are mediated by both metabolic and environmental cues. Inflammatory bowel disease (IBD) is a chronic disease that affects both adults and children alike, imposing significant morbidity and healthcare costs. Understanding how the intestinal epithelium self-renews and maintains its integrity in the face of inflammatory damage is key to the development of novel therapies for intestinal disease. My ultimate goal is to head an independent research program focused on translational intestinal disease models and therapeutic design. Career goals: Through the framework delineated by this proposal, I will establish an independent research program dedicated to understanding intestinal pathophysiology and the development of novel therapeutics. The research plan described herein is designed to expand my expertise into new arenas of animal genetics and disease modeling. This is consistent with my long-term goals of translational science, bringing discoveries from my laboratory back to benefit my patients. The mentoring I will receive by my Advisory Committee coupled with the career development workshops at UCSF such as grant and manuscript writing, will foster both my professional and scientific development. Environment: I have identified a strong mentoring team lead by Dr. Holly Ingraham, a leader in nuclear receptor biology. Dr. Averil Ma, an expert in IBD and animal models of colitis, and Dr. Ophir Klein, a physician scientist with epithelial stem cell expertise, round out my Advisory Committee with complementary skill sets. To enhance my research training, I will engage in didactic coursework in immunology, advanced light microscopy, high throughput data analysis, and the responsible conduct of research. UCSF provides a stimulating environment for the pursuit of science with a highly collaborative network of investigators. The Department of Pediatrics and the Division of Gastroenterology each have a long history of developing future leaders in basic and clinical science. In summary, as a highly motivated clinician scientist, my ultimate goal is to develop an independent basic and translational research program focused on intestinal disorders. This proposal provides necessary training in research coupled with a strong mentoring plan to help me achieve success.