Targeting Intestinal Epithelium to Promote Healing in Inflammatory Bowel Disease

针对肠上皮促进炎症性肠病的愈合

基本信息

批准号：
9180524
负责人：
James Bayrer
金额：
$ 15.86万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-07-18 至 2019-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9180524
关键词：
Address Adult Advisory Committees Affect Animal Disease Models Animal Genetics Animal Model Animals Anti-Inflammatory Agents Anti-inflammatory Apoptosis Back Basic Science Binding Biology Breeding Cells ChIP-seq Child Childhood Chronic Chronic Disease Clinical Clinical Sciences Colitis Complement Coupled Cues Data Data Analyses Dependovirus Development Diagnosis Disease Disease model Dissection Drug Targeting Educational workshop Engineering Environment Epithelial Epithelial Cells Experimental Models Fostering Functional disorder Future Gastroenterologist Gastroenterology Gastrointestinal tract structure Gene Expression Gene Targeting Genes Genetic Models Genetic Transcription Goals Grant Growth Head Healed Health Care Costs Holly Homeostasis Human Immune Immunology Individual Inflammation Inflammatory Inflammatory Bowel Diseases Inflammatory Response Inflammatory disease of the intestine Injury Intestinal Diseases Intestines Knock-out Knockout Mice Laboratories Lead Ligand Binding Ligands LoxP-flanked allele Manuscripts Mediating Mentors Metabolic Modeling Molecular Morbidity - disease rate Mus Mutation NR5A2 gene Natural regeneration Nuclear Receptors Organoids Pathway interactions Patients Pharmacology Physicians Predisposition Production Recording of previous events Research Research Personnel Research Training Resistance Review Committee Role Science Scientist Speed Staging Stem cells Steroids Suggestion Sulfonic Acids System Techniques Testing Therapeutic Tight Junctions Training Transcript Translational Research Trinitrobenzenes Work Writing base career career development cell growth design drug development genetic manipulation healing humanized mouse immune function in vivo in vivo Model inflammatory marker interest intestinal crypt intestinal epithelium light microscopy mouse model mutant new therapeutic target novel novel therapeutics overexpression pediatric department programs receptor response response to injury responsible research conduct self-renewal skills success tissue culture transcriptome sequencing villin

项目摘要

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.

项目概要/摘要方法：我将重点关注越来越被认可的 NR5A 核受体肝受体同源物-1（LRH-1、NR5A2）因其在细胞生长中的作用及其抗炎转录谱。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 的丢失会损害上皮更新和对损伤的反应，并且增强的 LRH-1 表达会抑制肠道炎症并加速粘膜愈合。我开发了一种离体肠道类器官培养物（肠）系统研究上皮对损伤的反应。 Enteroid 系统允许我直接解决通过使用条件小鼠遗传模型消除 LRH-1 在肠上皮病理生理学中的作用肠上皮细胞中的 LRH-1 (Villin-CreER)。我已经调整了腺相关病毒（AAV）系统使人类LRH-1的表达和肠样系统中重要的LRH-1突变能够进一步探测 LRH-1 介导的上皮功能。我将使用具有条件 LRH-1 敲除和过表达以我的离体研究为基础，并定义 LRH-1 在上皮损伤和炎症反应。我的工作将确定 LRH-1 激活是否足以削弱免疫介导粘膜损伤并询问 LRH-1 是否可以作为 IBD 药物开发的目标。我相信问题是这个基于指导的 K08 应用程序中概述的实验模型令人兴奋且高度与我对炎症性肠道疾病的临床兴趣相关。候选人及理由：我是一名双学位医师科学家 MSTP 毕业生，接受过胃肠病学和药理学培训。在这个我寻求了解控制肠道稳态和适应性的分子途径驱动损伤反应的机制。在肠道中，炎症反应由两者介导代谢和环境线索。炎症性肠病 (IBD) 是一种影响两种疾病的慢性疾病成人和儿童都一样，造成巨大的发病率和医疗费用。了解如何肠上皮在面对炎症损伤时自我更新并保持其完整性是关键开发肠道疾病的新疗法。我的最终目标是领导一项独立研究项目重点关注转化肠道疾病模型和治疗设计。职业目标：通过这个提案所描绘的框架，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，即从我的实验室回来造福我的病人。我将获得咨询委员会的指导以及加州大学旧金山分校的职业发展研讨会，例如资助和手稿写作，将培养我的专业化、科学化发展。环境：我已经确定了一支由核受体生物学领域的领军人物 Holly Ingraham 博士领导的强大指导团队。博士。 Averil Ma（IBD 和结肠炎动物模型专家）和 Ophir Klein 博士（医学科学家）上皮干细胞专业知识，使我的顾问委员会拥有互补的技能。为了增强我的研究培训，我将从事免疫学、高级光学显微镜、高吞吐量数据分析，以及负责任的研究行为。 UCSF 提供了一个激励性的环境通过高度协作的研究人员网络追求科学。儿科和胃肠病学部门在培养基础和临床领域的未来领导者方面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，我的最终目标是发展一种独立的基础和专注于肠道疾病的转化研究计划。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成功。