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: 9034167
• 负责人: Natalie A Terry
• 金额: $ 15.08万
• 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/9034167
• 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; 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; glucagon-like peptide 1; gut microbiota; intestinal epithelium; loss of function; medical schools; meetings; member; mortality; mouse model; overexpression; programs; promoter; public health relevance; 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(申请人提供)：肠源性内分泌缺乏症是一种先天性腹泻疾病，由肠道内分泌细胞丧失引起。除了支持性治疗外，这种疾病和其他与肠道内分泌细胞发育不良相关的腹泻疾病没有治疗选择。这项建议集中在诱导性肠道Arx缺乏症(ArxIKO)的小鼠模型上，以了解这些疾病中吸收不良的机制。肠道Arx缺乏会导致肠道内分泌发育受损，表现为腹泻、脂肪吸收不良和粪便短链脂肪酸受损。初步数据表明，低脂饮食可以恢复肠道Arx缺乏小鼠的正常生长，这意味着低脂饮食干预是一种 治疗选择。概述的具体目标将描述ArxIKO小鼠的肠道内分泌细胞的谱系分配，以及在低脂肪(来自脂肪的10%卡路里)和高脂肪(来自脂肪的45%热量)饮食中的生长。在挑战高脂肪饮食的过程中，我将用质谱学和比色法仔细研究Arxint小鼠的低温过程。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，我将在低脂和高脂饮食下对肠道Arx消融后的肠道微生物区系进行表征，测定粪便短链脂肪酸含量，并进行粪便移植，以确定微生物区系在疾病发病机制中的作用。直接下游靶点将通过CHIP-SEQ鉴定，在荧光素酶检测中验证，并在肠道类器官培养中过表达，以挽救肠道内分泌谱系。最后，将在肠道器官中建立脂类吸收试验，以验证二级靶点。我的建议概述了一个为期3年的培训计划，以使我作为一名儿科胃肠病学的内科科学家继续发展。作为一名临床医生，我在先天性腹泻方面积累了专业知识，并成为我们肠道康复计划的一部分。作为一名科学家，我有一位杰出的导师克劳斯·凯斯特纳博士，他非常慷慨地花费了他的时间和资源。凯斯特纳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，我有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内科科学家，包括宾夕法尼亚大学的Gary Wu博士(成人胃肠病学家)和Doris Stoffers博士(成人内分泌学家)，以及费城儿童医院的Eric Marsh博士(儿科神经科医生)和Robert Heuckeroth博士(儿科胃肠病学家)。此外，与丹尼尔·雷德博士一起接受脂质生物学领域的培训将加深我对脂质吸收不良的认识。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相结合，为我作为一名内科科学家的发展创造了一个理想的环境，因为我继续在儿科胃肠病领域做出贡献，并在提交R01报告方面取得进展。