Investigation of Regional Identity in Human Intestinal Stem Cells

描述（由申请人提供）：小肠在其整个长度上具有相似的隐窝和绒毛形态，但功能上的显著区域差异是众所周知的。 这些差异的临床相关性通过许多人类疾病在整个肠道中的区域特异性位置来突出。了解导致区域特异性肠干细胞（ISC）的机制，需要开发恢复区域特异性功能的治疗方法。我们的初步数据支持这样的概念，即肠的模式化发生在发育的早期，并在整个生命过程中维持在ISC的水平。使用我们最近开发的体外方法来培养人ISC衍生的类肠体（hENO）和区域模式化人ES/iPS衍生的类器官（HIO）的条件，结合独特的移植模型，我们将确定肠道区域身份（rID）是否由特定的转录因子以及单个ISC内独特的染色质状态稳定维持。为了验证这一假设，我们将定义肠道区域身份的分子和细胞载体，确定人类hENO和HIO在移植后是否保持稳定的rID，并确定区域身份的分子决定因素网络。

DESCRIPTION (provided by applicant): The small intestine bears similar morphology of crypts and villi throughout its length, yet marked regional differences in function are well known. The clinical relevance of these differences is highlighted by the region specific location of many human diseases throughout the bowel. Understanding the mechanisms that result in regional specific intestinal stem cells (ISC) is required to develop therapeutic approaches for restoring regional specific function. Our preliminary data supports the concept that patterning of the intestine occurs early in development and is maintained throughout life at the level of the ISC. Using our recently developed in vitro methods to culture human ISC-derived enteroids (hENO) and conditions that regionally pattern human ES/iPS-derived organoids (HIOs), combined with unique transplantation models, we will determine if intestinal regional identity (rID) is stably maintained by specific transcription factors together with a unique chromatin state within individual ISC. To test this hypothesis, we will define the molecular and cellular carriers of intestinal regional identity, determine whether human hENOs and HIOs maintain stable rID after transplantation and identify the network of molecular determinants of regional identity.