Targeting Intestinal Epithelium to Promote Healing in Inflammatory Bowel Disease

项目摘要/摘要 方法： 我将重点介绍NR5A核受体肝脏受体Homolog-1(LRH-1，NR5A2)，越来越多的人认识到 因为它在细胞生长和抗炎转录过程中的作用。缺乏肠道LRH-1基因敲除的小鼠 在IBD模型中显示黏膜损伤和炎性标志物增加。我假设LRH-1的丢失 会损害上皮细胞的更新和对损伤的反应，这种增强的LRH-1表达将抑制 肠道发炎，加速粘膜愈合。我已经开发了一种体外肠道有机培养方法 (肠样)系统，用于研究上皮细胞对损伤的反应。肠腔系统让我可以直接处理 利用条件性小鼠遗传模型研究LRH-1在肠上皮细胞病理生理中的作用 LRH-1在肠上皮细胞(绒毛膜-克里尔)。我已经将腺相关病毒(AAV)系统改造成 使人LRH-1在肠样系统中的表达和重要的LRH-1突变能够进一步探测 LRH-1介导的上皮功能。我将使用体内IBD模型，条件LRH-1基因敲除和 过度表达以我的体外研究为基础，并确定LRH-1在上皮损伤和 炎症反应。我的工作将确定LRH-1的激活是否足以钝化免疫介导的 并询问LRH-1是否可以作为IBD药物开发的靶点。我相信问题是 这款基于导师的K08应用程序中概述的问答和实验模型非常令人兴奋和高度 与我在炎症性肠病方面的临床兴趣有关。 候选人和理由： 我是一名双学位内科科学家，MSTP毕业生，受过胃肠病学和药理学的培训。在这 建议I试图了解控制肠道内稳态和适应性的分子通路 驱动伤害反应的机制。在肠道中，炎症反应是由两种介质介导的 新陈代谢和环境线索。炎症性肠病(IBD)是一种影响两端的慢性疾病。 成年人和儿童都是如此，造成了巨大的发病率和医疗费用。了解如何将 肠道上皮在面对炎性损伤时自我更新并保持其完整性是 肠道疾病新疗法的开发。我的最终目标是领导一项独立的研究 项目重点是转化性肠道疾病模型和治疗设计。 职业目标： 通过这项提议划定的框架，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开发。这个 这里描述的研究计划旨在将我的专业知识扩展到动物遗传学和动物遗传学的新领域 疾病模型。这与我对翻译科学的长期目标是一致的，从 带回我的实验室，让我的病人受益。我的咨询委员会将给我的指导加上 加州大学旧金山分校的职业发展研讨会，如助学金和手稿写作，将培养我的 专业化、科学化发展。 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霍莉·英格拉汉姆博士领导。Dr。 Averil Ma，IBD和结肠炎动物模型专家，以及Ophir Klein博士， 上皮干细胞专业知识，使我的咨询委员会具有互补的技能组合。为了增强我的 研究培训，我将从事免疫学、高级光学显微镜、高级 吞吐量数据分析，并负责任地进行研究。加州大学旧金山分校提供了一个刺激的环境 与高度合作的调查人员网络一起追求科学。儿科和医学系 消化内科在培养基础和临床方面的未来领导者方面都有着悠久的历史 科学。 总而言之，作为一名积极进取的临床科学家，我的最终目标是开发一种独立的基础和 翻译研究计划侧重于肠道疾病。该提案提供了必要的培训 研究加上强有力的指导计划，帮助我取得了成功。

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.