权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

A Novel In Vivo Mouse Model to Study Myofibroblast-Epithelial Cell Interactions

研究肌成纤维细胞-上皮细胞相互作用的新型体内小鼠模型

基本信息

批准号：
9262923
负责人：
James Yoo
金额：
$ 8.25万
依托单位：
TUFTS MEDICAL CENTER
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-05-01 至 2019-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9262923
关键词：
Animal Model Animals Architecture Bacteria Cell Communication Cell Culture Techniques Cell Differentiation process Cell Proliferation Cell physiology Cells Coculture Techniques Colitis Colon Colonoscopy Development Diffuse Distal Drug Delivery Systems Elements Environment Epithelial Cell Proliferation Epithelial Cells Epithelial-Stromal Communication Epithelium Event Fibrosis Gastrointestinal Diseases Gastrointestinal tract structure Gene Targeting Growth Harvest Immune Implant In Vitro Inflammation Inflammation Mediators Influentials Injectable Injection of therapeutic agent Intestinal Mucosa Intestines Lamina Propria Malignant Neoplasms Mediating Modeling Mus Myofibroblast Nerve Organ Pathologic Physiological Play Population Process Reproducibility Research Role Signal Pathway Signal Transduction Sodium Dextran Sulfate Source Stromal Cells Techniques Therapeutic Intervention Time Tissues Villous body system cell type cyclooxygenase 2 gastrointestinal healing implantation in vivo in vivo Model injury and repair migration minimally invasive mouse model novel paracrine protein kinase D public health relevance repaired stem stem cell niche subcutaneous tissue support frame tool

项目摘要

DESCRIPTION (provided by applicant): The myofibroblast is an influential stromal cell of the gastrointestinal (GI) tract that regulates many important processes ranging from cell proliferation and differentiation along the crypt-villous axis, mucosal repair, fibrosis, and the development of cancer [7-9]. The signaling pathways that regulate myofibroblast function have been studied in vitro [10, 11]. However, the actual physiologic impact of these signaling events on the overlying epithelium remains largely speculative due to limitations with existing co-culture and animal models. In vitro co-culture models lack the normal bowel wall architecture and surrounding cell populations that are essential elements of the GI microenvironment [9]. Animal models that utilize conditional gene targeting are neither organ nor cell type-specific since the myofibroblast lacks a unique cell marker [12, 13]. Current research efforts are limited by a paucity of in vivo models to effectively study how the myofibroblast regulates epithelial cell function under normal and pathologic conditions. This proposal involves the development of a novel technique utilizing murine colonoscopy that will allow for the real-time in vivo study of myofibroblast-epithelial cell interactions. The technique first involves the isolation of primary myofibroblasts from mouse colon tissue and subsequent growth in cell culture. Primary myofibroblasts will then be re-implanted endoscopically by injection into the colon wall of an immune-competent, syngeneic mouse. The technique allows for targeted manipulation of a subpopulation of myofibroblasts in a segment of mouse colon that can be easily re-identified, serially evaluated, and directly compared to adjacent tissue under the same experimental conditions. This technique can be applied to any existing mouse model to study stromal-epithelial interactions in the distal colon. The proposal aims to 1) demonstrate that myofibroblasts can be successfully and reproducibly implanted in the colon wall and can maintain viability in live, immune-competent, syngeneic mice, and as a proof of principle, that 2) this model can be used to study stromal-epithelial cell interactions, specifically in the context of an animal model of injury-repair.

描述（由申请人提供）：肌成纤维细胞是胃肠道（GI）中一种有影响力的基质细胞，调节从细胞增殖等许多重要过程以及沿隐窝-绒毛轴的分化、粘膜修复、纤维化和癌症的发展[7-9]。调节肌成纤维细胞功能的信号通路已在体外进行了研究 [10, 11]。然而，由于现有共培养和动物模型的局限性，这些信号事件对上皮的实际生理影响在很大程度上仍然是推测性的。体外共培养模型缺乏正常的肠壁结构和周围细胞群，而这些是胃肠道微环境的基本要素[9]。利用条件基因靶向的动物模型既不具有器官特异性，也不具有细胞类型特异性，因为肌成纤维细胞缺乏独特的细胞标记 [12, 13]。目前的研究工作受到体内模型缺乏的限制，无法有效研究肌成纤维细胞在正常和病理条件下如何调节上皮细胞功能。该提案涉及开发一种利用小鼠结肠镜检查的新技术，该技术将允许对肌成纤维细胞上皮细胞进行实时体内研究互动。该技术首先涉及从小鼠结肠组织中分离原代肌成纤维细胞，然后在细胞培养物中生长。然后通过内窥镜注射到具有免疫能力的同基因小鼠的结肠壁中，将原代肌成纤维细胞重新植入。该技术允许对小鼠结肠一段中的肌成纤维细胞亚群进行有针对性的操作，可以轻松地重新识别、连续评估，并在相同的实验条件下直接与相邻组织进行比较。该技术可应用于任何现有的小鼠模型，以研究远端结肠中的基质-上皮相互作用。该提案的目的是 1) 证明肌成纤维细胞可以成功且可重复地植入结肠壁，并且可以在活的、具有免疫能力的同基因小鼠中维持活力，并且作为原理证明，2) 该模型可用于研究基质-上皮细胞相互作用，特别是在损伤修复动物模型的背景下。