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（由申请人提供）：因克罗恩病、缺血、创伤或放射性肠炎等疾病而进行手术切除后功能性小肠表面积的丧失可能导致短肠综合征（SBS），这是发病率、死亡率和医疗保健的重要原因美国 SBS 患者经常依赖肠外营养来满足其营养需求，在肠切除后，剩余的小肠上皮会产生适应性反应，增加绒毛高度和隐窝。然而，人类的适应性反应是不可预测的，即使有对残余小肠长度的准确评估，也可能需要长达两年的时间才能确定哪些患者将停止肠外营养。这种临床变异性是未知的。我们的应用旨在利用 PI 的大量 SBS 患者群体的患者活检来探索新的再生治疗方法，以增强小肠功能表面积。我们的方法将转化我们的短肠综合征临床前模型的观察结果。干细胞微环境中的肌成纤维细胞信号转导具有治疗潜力的人类 SBS 研究表明，肌成纤维细胞蛋白 Epimorphin (Epim) 与 Epim-/ 共培养可调节隐窝裂变、肠上皮增殖和肌成纤维细胞分泌功能。 - 肌成纤维细胞进一步增强 WT 小鼠隐窝肠样生长和表面积，我们的首要目标是增强增殖上皮反应并增加小肠。我们的总体假设是，调节肌成纤维细胞 Epim 及其靶标将增强人类小肠生长，为 SBS 提供一种新颖的再生治疗方法，目标 1 将通过建立干细胞/肠样蛋白直接转化我们的小鼠研究。我们将解决与 Epim 缺陷的肌成纤维细胞共培养是否会增加人肠样细胞生长的假设。体外肠样细胞的生长和增殖能力可以预测肠外营养的适应能力和断奶能力。目标 2 将解决两个假设：1. Epim 缺失可调节 MF-隐窝上皮相互作用，从而通过改变来增加隐窝干细胞增殖和肠样细胞生长。 Bmp、Hh 和 Wnt 信号通路在调节 MF 分泌中表现出特定的抑制作用和促胞吐作用。我们从临床前研究到 Epim 缺失对干细胞生态位中 MF 功能影响的人体研究中的发现，我们将扩大我们对 SBS 患者小肠隐窝干/肠样培养物的独特临床收集，并为未来潜在的高通量生成国家资源。筛选增强SBS中干细胞和隐窝细胞增殖的治疗剂以及通过Epim缺失调节的可溶性因子的分泌代表了促进肠干/隐窝上皮增殖、导致适应的潜在新治疗靶点。