MYOFIBROBLAST REGULATION OF THE STEM CELL NICHE IN SHORT BOWEL SYNDROME

短肠综合征中肌成纤维细胞对干细胞生态位的调节

• 批准号: 9306091
• 负责人: DEBORAH C. RUBIN
• 金额: $ 34.31万
• 依托单位: WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-09-01 至 2019-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9306091
• 关键词: Address; Area; Biology; Biopsy; Cell Communication; Cell Count; Cell Culture Techniques; Cell Proliferation; Clinical; Coculture Techniques; Collaborations; Collection; Crohn' s disease; Data; Development; Differentiation and Growth; Disease; Electrolytes; Endoscopic Biopsy; Environment; Epithelial; Epithelium; Erinaceidae; Excision; Future; Goals; Growth; Health Care Costs; Height; Human; In Vitro; Injury; Intestines; Ischemia; Length; Measurement; Mediating; Morbidity - disease rate; Mus; Myofibroblast; Necrotizing Enterocolitis; Nutrient; Nutritional Requirements; Operative Surgical Procedures; Organoids; Parenteral Nutrition; Pathway interactions; Patients; Play; Pre-Clinical Model; Proteins; Radiation Enteritis; Regulation; Resources; Role; Short Bowel Syndrome; Signal Pathway; Signal Transduction; Small Intestines; Stem cells; Surface; Therapeutic; Therapeutic Agents; Translating; Trauma; Villus; WNT Signaling Pathway; Weaning; absorption; cohort; crypt cell; epimorphin; functional loss; high throughput screening; in vivo; mortality; new therapeutic target; novel; patient population; preclinical study; public health relevance; regenerative therapy; response; smoothened signaling pathway; stem; stem cell niche; syntaxin; virtual

 DESCRIPTION (provided by applicant): Loss of functional small bowel surface area following surgical resection for disorders including Crohn's disease, ischemia, trauma, or radiation enteritis may result in short bowel syndrome (SBS), an important cause of morbidity, mortality and health care costs in the U.S. SBS patients are frequently dependent on parenteral nutrition to meet their nutritional requirements. Following intestinal resection, the remaining small bowel epithelium mounts an adaptive response that increases villus height, crypt depth and enhances nutrient and electrolyte absorption. However, the adaptive response in humans is unpredictable and it may require up to two years to determine which patients will wean off parenteral nutrition, even with an accurate assessment of remnant small bowel length. The mechanistic basis for this clinical variability is unknown. Our application seeks to explore new regenerative, therapeutic approaches to enhance small bowel functional surface area, using patient biopsies from the PI's large SBS patient population. Our approach will translate observations from our preclinical models of short bowel syndrome and myofibroblast signaling in the stem cell niche into studies in human SBS that have therapeutic potential. We have shown that a myofibroblast protein, epimorphin (Epim) regulates crypt fission, gut epithelial proliferation and myofibroblast secretory function in mice. Co-culture with Epim-/- myofibroblasts further enhances WT mouse crypt enteroid growth and surface area. Our overarching goals are to enhance the proliferative epithelial response and increase small bowel surface area as a therapeutic strategy for SBS. Our overarching hypothesis is that modulating myofibroblast Epim and its targets will enhance human small bowel growth providing a novel regenerative therapeutic approach to SBS. Aim 1 will directly translate our mouse studies by establishing stem cell/enteroid and myofibroblast cultures from the PI's large cohort of SBS patients. We will address the hypothesis that co- culture with Epim-deficient myofibroblasts increases the growth of human enteroids. We will determine whether growth and proliferative capacity of enteroids in vitro can predict the ability t adapt and wean from parenteral nutrition. Aim 2 will address two hypotheses. 1. MF-crypt epithelial interactions are modulated by Epim deletion to increase crypt stem cell proliferation and enteroid growth via altered Bmp, Hh, and Wnt signaling pathways. 2. Epim demonstrates context-specific inhibitory vs. pro-exocytotic roles in regulating MF secretion. Our studies are significant because we will directly translate our findings from preclinical studies to human studies of Epim deletion's effects on MF function in the stem cell niche. We will expand our unique clinical collection of small bowel crypt stem/enteroid cultures from SBS patients, and generate a national resource for potential future high throughput screening of therapeutic agents that enhance stem and crypt cell proliferation in SBS. Epim and the secretion of soluble factors that are modulated by Epim deletion represent potential novel therapeutic targets to promote intestinal stem/crypt epithelial proliferation, leading to adaptation. .

 描述（由适用提供）：手术切除后功能性小肠表面积的损失包括克罗恩病，缺血，创伤或放射性肠炎，可能导致短肠综合征（SBS）短肠综合征（SBS），这是美国SBS患者的发病，死亡率和医疗保健成本的重要原因，通常依赖于父母的营养需求。肠切除后，其余的小肠上皮会安装自适应反应，从而增加绒毛的高度，加密深度并增强营养和电解质的痛苦。但是，人类的自适应反应是不可预测的，即使对残留的小肠长度进行了准确的评估，也可能需要长达两年的时间才能确定哪些患者会脱离父母的营养。该临床变异性的机械基础尚不清楚。我们的应用程序旨在探索新的再生治疗。使用PI大型SBS患者人群的患者活检，可以增强小肠功能表面积的方法。我们的方法将使我们的短肠综合征和干细胞生态裂细胞信号传导的临床前模型转化为具有治疗潜力的人类SB的研究。我们已经表明，肌纤维细胞蛋白（EPIM）调节小鼠的隐窝裂变，肠道上皮增殖和肌纤维细胞秘书功能。与EPIM - / - 肌纤维细胞共同培养进一步增强了WT小鼠加密型肠内生长和表面积。我们的总体目标是增强上皮反应，并增加小肠表面积作为SBS的治疗策略。我们的总体假设是，调节肌纤维细胞EPIM及其靶标将增强人类小肠的生长，从而为SBS提供一种新型的再生治疗方法。 AIM 1将通过建立PI大量SBS患者队列的干细胞/肠细胞和肌纤维细胞培养物来直接转化我们的小鼠研究。我们将解决以下假设：与EPIM缺陷型成肌纤维细胞共同培养会增加人类肠动物的生长。我们将确定体外肠动物的生长和增殖能力是否可以预测父母营养的适应能力和we依。 AIM 2将解决两个假设。 1。通过EPIM缺失调节MF-Crypt上皮相互作用，以通过改变的BMP，HH和Wnt信号通路来增加加密细胞增殖和肠托的生长。 2。上皮相互作用通过EPIM缺失调节，以通过改变的BMP，HH和WNT信号通路来增加加密细胞增殖和肠托的生长。我们的研究很重要，因为我们将直接将我们的发现从临床前研究转化为对EPIM缺失对干细胞生态位MF功能的影响的人类研究。我们将扩大来自SBS患者的小肠隐窝茎/肠培养物的独特临床收集，并为潜在的未来对治疗剂的高吞吐量筛查生成国家资源，从而增强SBS中的茎和隐层细胞增殖。 EPIM和由EPIM缺失调节的固体因子的分泌代表了潜在的新型治疗靶标，以促进肠干/隐窝上皮增生，从而导致适应。