ROLE OF INTESTINAL ARX IN ENTEROENDOCRINE DEVELOPMENT AND CONGENITAL DIARRHEA

肠道 ARX 在肠内分泌发育和先天性腹泻中的作用

• 批准号: 9147576
• 负责人: Natalie A Terry
• 金额: $ 14.93万
• 依托单位: CHILDREN'S HOSP OF PHILADELPHIA
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-09-30 至 2018-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9147576
• 关键词: Ablation; Adult; Advisory Committees; Age; Biological Assay; Biology; Butyrates; Calories; Cell Lineage; Cells; ChIP-seq; Childhood; Cholecystokinin; Communities; Congenital Disorders; Data; Desire for food; Development; Diarrhea; Diet; Dietary Intervention; Digestion; Disease; Education; Endocrine; Endocrine system; Endocrinologist; Endoderm; Enteral; Enterocytes; Enteroendocrine Cell; Environment; Exhibits; Exocrine pancreatic insufficiency; Failure; Failure to Thrive; Fat-Restricted Diet; Fatty acid glycerol esters; Feces; Functional disorder; Gastrins; Gastroenterologist; Gastroenterology; Genetic; Germ-Free; Growth; Health; High Fat Diet; Histologic; Hormonal; Hormones; Immunohistochemistry; Infant; Intestinal Diseases; Intestinal Hormones; Intestines; Intravenous; Knowledge; Lead; Lipids; Luciferases; Maintenance; Malabsorption Syndromes; Mass Spectrum Analysis; Mediating; Mentors; Metabolism; Modeling; Molecular Profiling; Mus; Mutation; Neurologist; Nutrient; Nutritional Support; Organoids; Pancreas; Pancreatic enzyme; Pathogenesis; Patients; Pediatric Hospitals; Pediatrics; Pennsylvania; Phenotype; Philadelphia; Physicians; Physiology; Process; Propionates; Public Health; Publications; Rare Diseases; Rehabilitation therapy; Research; Resources; Role; Scientist; Signal Transduction; Somatostatin; Supplementation; Supportive care; System; Testing; Therapeutic; Time; Training; Training Programs; Transplantation; Universities; Volatile Fatty Acids; absorption; blood glucose regulation; career development; cell motility; drinking water; early onset; fatty acid metabolism; fecal transplantation; glucagon-like peptide 1; gut microbiota; intestinal epithelium; loss of function; medical schools; meetings; member; microbiota; mortality; mouse model; overexpression; programs; promoter; rRNA Genes; research study; response; villin

 DESCRIPTION (provided by applicant): Enteric anendocrinosis is a congenital diarrheal disorder caused by the loss of the intestinal endocrine cells. Aside from supportive care, this disorder and other related diarrheal disorders with enteroendocrine cell dysgenesis do not have therapeutic options. This proposal focuses on a mouse model of inducible intestinal Arx deficiency (ArxIKO) to understand the mechanism of malabsorption in these disorders. Intestinal Arx deficiency causes impaired enteroendocrine development with diarrhea, lipid malabsorption, and impaired fecal short chain fatty acids. Preliminary data suggests that a low fat diet restores normal growth in the intestinal Arx-deficient mice, implicating a low fat dietary intervention as a therapeutic option. The specific aims outlined will characterize the lineage allocation of enteroendocrine cells in ArxIKO mice and growth on low-fat (10% calories from fat) and high-fat (45% calories from fat) diet. During challenge with a high-fat diet, I will carefully investigate lpid processing in the Arxint mice with both mass spectrometry and colorimetric assays. Ultimately, pancreatic enzyme supplementation will be provided to understand the contribution of secondary pancreatic insufficiency. As a new aim, I will characterize the intestinal microbiota in response to intestinal Arx ablation on the low-fat and high-fat diet, determine the fecal short chain fatty acid content, and perform fecal transplantation to determine the contribution of the microbiota to the pathogenesis of disease. Direct downstream targets will be identified with ChIP-seq, validated in luciferase assays, and overexpressed in intestinal organoid culture in order to rescue enteroendocrine lineages. Finally, a lipid absorption assay will be developed in the intestinal organoids to validate secondary targets. My proposal outlines a 3-year training program for my continued development as a physician-scientist in Pediatric Gastroenterology. As a clinician, I have developed an expertise in congenital diarrhea and am part of our Intestinal Rehabilitation Program. As a scientist, I have an extraordinary mentor in Dr. Klaus Kaestner who has been very generous with his time and resources. The Kaestner lab is also a vibrant community for scientific development. To further promote my career development, I have a Scientific Advisory Committee with whom I will meet twice a year. All members are highly regarded physician-scientists and include Dr. Gary Wu (adult gastroenterologist) and Dr. Doris Stoffers (adult endocrinologist) at the University of Pennsylvania and Dr. Eric Marsh (pediatric neurologist) and Dr. Robert Heuckeroth (pediatric gastroenterologist) at The Children's Hospital of Philadelphia. Additionally, training with Dr. Daniel Rader in the field of lipid biology will exand my education on lipid malabsorption. The combination of the research community and state-of-the-art facilities at The Children's Hospital of Philadelphia and the Perelman School of Medicine at the University of Pennsylvania create an ideal environment for my development as a physician-scientist as I continue to make contributions to the field of pediatric gastroenterology and progress toward an R01 submission.

 描述（由申请人提供）：肠道无内分泌病是一种由肠道内分泌细胞丢失引起的先天性肠道疾病。除了支持性治疗，这种疾病和其他相关的肠道疾病与肠内分泌细胞发育不全没有治疗选择。该提案的重点是诱导型肠Arx缺乏症（ArxIKO）的小鼠模型，以了解这些疾病中吸收不良的机制。肠Arx缺乏导致肠内分泌发育受损，伴有腹泻、脂质吸收不良和粪便短链脂肪酸受损。初步数据表明，低脂肪饮食可以恢复肠道Arx缺陷小鼠的正常生长，这意味着低脂肪饮食干预是一种有效的方法。 治疗选择 概述的具体目标将表征ArxIKO小鼠中肠内分泌细胞的谱系分配以及低脂肪（10%来自脂肪的卡路里）和高脂肪（45%来自脂肪的卡路里）饮食的生长。在高脂饮食的挑战过程中，我将用质谱和比色分析仔细研究Arxint小鼠的脂质加工。最终，将提供胰酶补充剂以了解继发性胰腺功能不全的作用。作为一个新的目标，我将描述肠道微生物群对低脂肪和高脂肪饮食的肠道Arx消融的反应，确定粪便短链脂肪酸含量，并进行粪便移植以确定微生物群对疾病发病机制的贡献。直接下游靶标将用ChIP-seq鉴定，在荧光素酶测定中验证，并在肠类器官培养物中过表达，以拯救肠内分泌谱系。最后，将在肠道类器官中开发脂质吸收测定法，以验证次要靶标。 我的建议概述了一个为期3年的培训计划，为我继续发展作为一个医生，科学家在儿科胃肠病学。作为一名临床医生，我在先天性腹泻方面积累了丰富的经验，并且是我们肠道康复计划的一部分。作为一名科学家，我有一位非凡的导师克劳斯·凯斯特纳博士，他非常慷慨地投入了时间和资源。Kaestner实验室也是一个充满活力的科学发展社区。为了进一步促进我的职业发展，我成立了一个科学咨询委员会，我每年与该委员会会面两次。所有成员都是备受推崇的医学科学家，包括宾夕法尼亚大学的加里吴博士（成人胃肠病学家）和多丽丝·斯托弗博士（成人内分泌学家）以及费城儿童医院的埃里克·马什博士（儿科神经学家）和罗伯特·赫克鲁斯博士（儿科胃肠病学家）。此外，与丹尼尔雷德博士在脂质生物学领域的培训将exand我的教育对脂质吸收不良。费城儿童医院和宾夕法尼亚大学佩雷尔曼医学院的研究社区和最先进的设施的结合为我作为一名医生科学家的发展创造了理想的环境，因为我继续为儿科胃肠病学领域做出贡献并向R 01提交进展。