Enteroendocrine cell reprogramming during intestinal injury

肠道损伤期间肠内分泌细胞重编程

• 批准号: 10573165
• 负责人: Brian David Muegge
• 金额: --
• 依托单位: ST. LOUIS VA MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-02-01 至 2025-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10573165
• 关键词: Academic skills; Achievement; Address; Adopted; Advisory Committees; Air; Animal Model; Animals; Applications Grants; Area; Award; Biology; Cell Culture System; Cell Culture Techniques; Cell Line; Cell Lineage; Cell Reprogramming; Cell physiology; Cell secretion; Cells; Cellular Stress; Colon; Computational Biology; Computer Analysis; Computing Methodologies; Data; Development; Developmental Biology; Disease; Doctor of Medicine; Doctor of Philosophy; Endocrine; Endocrine system; Endocrinology; Ensure; Enteroendocrine Cell; Epithelium; Fellowship; Foundations; Future; GLP-2; Genetic Transcription; Genomics; Goals; Goblet Cells; Grant; Growth; Growth Factor; Health Priorities; Healthcare Systems; Homeostasis; Hormone secretion; Hormones; In Vitro; Inflammatory Bowel Diseases; Injury; Internal Medicine; Intestinal Diseases; Intestines; Knock-out; Knowledge; Laboratories; Liquid substance; Medical center; Mentors; Metabolic Diseases; Metabolism; Methods; Modeling; Molecular; Monitor; Mucous Membrane; Outcome; Paracrine Communication; Pathway interactions; Patients; Pattern; Phenotype; Physicians; Population; Prevalence; Production; Publishing; Regulation; Research; Residencies; Resistance; Resolution; Sampling; Scientist; Serotonin; Signal Transduction; Somatostatin; Source; Structure; System; Techniques; Technology; Testing; Training; Universities; Veterans; Washington; Work; Writing; career; career development; cytokine; endoplasmic reticulum stress; epithelial injury; experience; experimental study; glucagon-like peptide 1; healing; in vitro Model; in vivo; injury and repair; insight; intercellular communication; intestinal epithelium; intestinal injury; microbial products; monolayer; new therapeutic target; novel; novel strategies; progenitor; programs; receptor; repaired; response; response to injury; single nucleus RNA-sequencing; single-cell RNA sequencing; skills; stem cells; symposium; transcriptomics; translational therapeutics; wound healing

RESEARCH STRATEGY: The primary research objective of this proposal is to address how subtypes of enteroendocrine cells change their activity (secretion) or state of differentiation in response to injury signals. A better understanding of intestinal injury repair is critical to VA health priorities; the prevalence of inflammatory bowel disease (IBD) has more than doubled among VA patients since 1998 and at least half of patients won’t experience full mucosal healing with existing therapies. Hormones produced in intestinal enteroendocrine cells (EEC) are altered in patients with IBD and can promote epithelial healing, but there is little understanding of how EEC secretion is regulated during injury. The study of EECs has been limited because they are rare, heterogenous, and not amenable to long-term primary culture with existing methods. To address this gap, we have developed a new experimental system to culture EECs from primary colon samples in long-term monolayers that supports all known EEC subtypes. We have discovered that subtypes of EEC are dynamically altered with a simple experimental injury in vitro. The altered EECs increase the expression of factors that promote intestinal healing. The goal of this proposal is to determine the cellular and transcriptional basis of how subtypes of EEC respond to intestinal injury through altered activity or differentiation. Our Aims are to (1) test the hypothesis that subtypes of EECs are adaptively reprogrammed by injury induced endoplasmic reticulum (ER) stress, (2) test the hypothesis that microenvironmental injury signals reprogram EECs to increase hormone secretion, and (3) define the transcriptional mechanisms of EEC reprogramming during injury at single-cell resolution. The expected outcome of this work is to establish the new paradigm that EECs undergo adaptive reprogramming during injury which will serve as the foundation for future independent grant proposals. Additionally, we expect to discover novel secreted factors or altered states of differentiation in EEC that would lead to new potential treatments for veterans suffering from intestinal or metabolic disorders. CANDIDATE/ENVIRONMENT: Dr. Brian Muegge is a senior fellow in the Division of Endocrinology at Washington University in St. Louis and the St. Louis VA Medical Center. He completed his M.D., Ph.D. training at Washington University where he performed graduate studies in Dr. Jeffrey Gordon’s laboratory. Dr. Muegge completed his residency in internal medicine at UCSF. He is conducting his fellowship research in the lab of his primary mentor Dr. Thaddeus Stappenbeck and co-mentor Dr. Carlos Bernal-Mizrachi at Washington University. Dr. Muegge has gained experience in intestinal stem cell culture and computational analysis of transcriptional data. He has developed a novel culture system that allows him to model EEC development and injury using primary cells. He now seeks to expand his expertise in new areas including lineage tracing, cell- cell signaling, and single cell RNA sequencing prior to starting his independent research career. CAREER DEVELOPMENT: This award will ensure that Dr. Muegge is able to establish a career as an independent physician scientist in the VA health care system. He has developed a formal 5-year training plan that will expand his scientific skills in intestinal development, in vitro models, and single cell transcriptomics, as well as academic skills in scientific publishing, grant writing, and laboratory management. Dr. Muegge’s training will be accomplished with experimental research, didactics, seminars, and national conferences. He has assembled a scientific advisory committee with the requisite experience and expertise to monitor and support his progress towards achieving his career goals. At the completion of the CDA-2, Dr. Muegge will be equipped with skills in cutting edge technology (single cell transcriptomics, in vitro stem cell culture systems) working in a unique area that sets him apart from others in the field (intestinal endocrine function during bowel injury). He will be uniquely prepared to address fundamental questions that require knowledge of intestinal biology, endocrinology, and computational biology.

研究策略：该提案的主要研究目标是解决该亚型的子类型 肠内分泌细胞因响应损伤信号而改变其活性（分泌）或分化状态。一个 更好地了解肠道损伤对VA健康的重点至关重要；炎症的患病率 自1998年以来，VA患者的肠道病（IBD）的两倍以上，至少一半的患者不会 通过现有疗法体验全面的粘膜愈合。肠肠内分泌细胞产生的激素 IBD患者的（EEC）发生了变化，可以促进上皮愈合，但对 在受伤期间如何调节EEC分泌。 EEC的研究受到限制，因为它们很少 具有现有方法的异质，不适合长期原发性培养。为了解决这个差距，我们 已经开发了一种新的实验系统，以长期培养初级结肠样品的EEC 支持所有已知的EEC亚型的单层。我们发现EEC的亚型是动态的 通过简单的实验损伤在体外改变。 EEC的改变增加了因素的表达 促进肠道愈合。该提案的目的是确定 EEC的亚型如何通过改变活性或分化来应对肠道损伤。我们的目标是（1） 检验以下假设：EEC的亚型被损伤诱导的内质的自适应重新编程 网状（ER）应力，（2）测试微环境损伤信号重编程EEC的假设 增加骑马分泌物，（3）定义EEC重新编程的转录机制 单细胞分辨率的伤害。这项工作的预期结果是建立新的范式 EEC在受伤期间进行自适应重新编程，这将成为未来独立的基础 赠款建议。此外，我们期望发现新颖的分泌因素或分化状态的变化 EEC将为患有肠道或代谢疾病的退伍军人带来新的潜在治疗方法。 候选人/环境：Brian Muegge博士是内分泌科的高级研究员 圣路易斯的华盛顿大学和圣路易斯弗吉尼亚州医疗中心。他完成了医学博士博士学位训练 在华盛顿大学，他在杰弗里·戈登（Jeffrey Gordon）博士的实验室学习研究生。 Muegge博士 在UCSF完成了他的内科住所。他正在他的实验室进行奖学金研究 华盛顿的Thaddeus Stappenbeck博士和同事Carlos Bernal-Mizrachi博士 大学。 Muegge博士在肠道干细胞培养和计算分析方面获得了经验 转录数据。他开发了一种新颖的文化系统，使他能够建模EEC的开发和 使用原代细胞损伤。现在，他试图扩大自己在新领域的专业知识，包括谱系跟踪，细胞 - 在开始独立研究生涯之前，细胞信号传导和单细胞RNA测序。 职业发展：该奖项将确保Muegge博士能够建立职业 VA卫生保健系统中的独立物理科学家。他制定了正式的5年培训计划 这将扩大他在肠道发展，体外模型和单细胞转录组学方面的科学技能，作为 以及科学出版，赠款写作和实验室管理方面的学术技能。 Muegge博士 培训将通过实验研究，教学学，半手级和民族会议来完成。他 已经组建了一个科学咨询委员会，并拥有必要的经验和专业知识，以监视和 支持他在实现职业目标方面的进步。 CDA-2完成时，Muegge博士将是 配备了尖端技术的技能（单细胞转录组学，体外干细胞培养系统） 在一个独特的领域工作，使他与野外的其他领域区分开来（肠内肠内分泌功能 受伤）。他将做出独特的准备，以解决需要肠道知识的基本问题 生物学，内分泌学和计算生物学。