Intestinal Stem Cell Culture and Entero-Endocrine Lineage Development

描述(由申请人提供)：人类和小鼠肠道干细胞生物学的分子基础是一个积极研究的主题。本应用的目的是确定影响人肠干细胞与肠肌成纤维细胞相互作用的介质，以促进人小肠上皮细胞的再生。这是为儿童肠衰竭相关疾病开发新的细胞疗法的必要步骤。为了将这种治疗方法带到临床上，有必要回顾一下肠道上皮的原位特征。我们将重点评估人类肠道干细胞进行自我更新和分化的机制。具体地说，我们将评估R-响应蛋白和Wnt5A在维持这些细胞的干性方面的作用。我们还将评估Hedgehog信号以及氧张力和pH变化引起的应激调节肠道干细胞和肌成纤维细胞之间相互作用的机制。最后，我们将评估特定的层粘连蛋白和整合素亚型是否会改变肠道干细胞的生长和分化，以及它们是否会影响受体动物模型中的细胞植入。在这项研究完成后，我们预计我们将能够确定控制这些干细胞特性的介质，这将导致新的细胞疗法。

DESCRIPTION (provided by applicant): The molecular basis of intestinal stem cell biology in both humans and mice is a subject of active research. The goal of this application is to identify the mediators that influence the interaction of human intestinal stem cells with intestinal myofibroblasts in order to enhance the regeneration of human small bowel epithelium. This is a necessary step toward the development of novel cellular therapies for disorders associated with intestinal failure in children. To bring such therapies to the clinic, it will be essential to recapitulate the in situ characteristics of the intestinal epithelium. We will focus on assessing te mechanism by which human intestinal stem cells undergo self-renewal and differentiation. Specifically, we will assess the role of R-Spondin and WNT5a in maintaining stemness of these cells. We will also assess the mechanism by which Hedgehog signaling and the stress induced by alterations of oxygen tension and pH modulate the interaction between intestinal stem cells and myofibroblasts. Lastly, we will assess whether specific laminin and integrin isotypes alter intestinal stem cell growth and differentiation, and whether they affect cellular engraftment in recipient animal models. At the completion of this study, we anticipate that we will be able to define the mediators that control these stem cell characteristics, which will lead to novel cellula therapies.