Characterization of intestinal stem cells (research project)

描述(由申请人提供)：存在于相互作用的利基中的肠道干细胞已被广泛接受为肠上皮持续自我更新的来源。然而，尽管其他器官系统在识别干细胞群体和利用这一知识进行疾病治疗方面取得了实质性进展，但由于无法明确识别、分离和培养干细胞群体，肠道干细胞领域的进展一直受到阻碍。最近发现了一些在干细胞巢内表现出不同表达模式的假定的肠道干细胞标记物，这表明多个祖细胞池共存。有趣的是，这些肠道祖细胞群体可能属于干细胞层次，类似于特征良好的造血系统中的干细胞层次。在肠道干细胞生态位中定义类似的层次结构将使我们能够前瞻性地分离离散的种群，从而扩展我们对肠道干细胞动力学的知识。 我们的长期研究目标是了解在动态平衡和疾病中调节肠道干细胞行为的机制。为肠道干细胞联合会的这一研究项目提出的研究旨在检验这样一种假设，即在正常组织和肠癌的癌症干细胞中，离散的肠道干细胞或祖细胞群体存在于一个层次结构中。针对这些目标的研究旨在利用免疫组织化学分析和三维肠腺重建的蛋白质表达模式以及荧光激活细胞分选的细胞表面抗原表达谱来定义祖细胞群体。此外，还将产生针对隐窝内未分化细胞的新型单抗，以扩大有效分离肠道祖细胞和干细胞的可用资源。分离离散的肠道干细胞池的能力对于促进我们对肠道生物学的理解至关重要，而肠道生物学对于理解、分类和治疗肠道疾病至关重要。 公共卫生相关性：解锁肠道生物学并利用这一知识治疗肠道疾病的关键在于识别和分离成人肠道干细胞的能力。这个项目旨在了解这种细胞的功能，将为这些干细胞功能不正常的疾病提供洞察力，例如在肠癌中。这些研究有可能改善诊断，提供指导治疗，并设计肠道疾病的新疗法。

DESCRIPTION (provided by applicant): The existence of intestinal stem cells residing in an interactive niche has been well-accepted as the source of continual self-renewal of the intestinal epithelium. However, while other organ systems have appreciated substantial progress in identifying stem populations and exploiting this knowledge for therapeutic treatment of disease, advances in the intestinal stem cell field have been hampered by the inability to definitively identify, isolate, and grow the stem cell population. Recent identification of a number of putative intestinal stem cell markers displaying distinct expression patterns within the stem cell niche suggests that multiple progenitor pools co-exist. Intriguingly, these intestinal progenitor populations may belong to a stem cell hierarchy analogous to that in the well-characterized hematopoietic system. Defining a similar hierarchy within the intestinal stem cell niche will allow us to prospectively isolate discrete populations, thereby extending our knowledge of intestinal stem cell dynamics. Our long-range research goal is to understand the mechanisms regulating intestinal stem cell behavior in homeostasis and in disease. The studies proposed for this Research Project for the Intestinal Stem Cell Consortium are designed to test the hypothesis that discrete populations of intestinal stem or progenitor cells exist within a hierarchy in both normal tissue, and among cancer stem cells in intestinal cancer. Studies to address these goals are designed to define progenitor populations using protein expression patterns by immunohistochemical analyses and 3-dimensional intestinal crypt reconstruction along with profiling of cell surface antigen expression by fluorescence-activated cell sorting. Further, novel monoclonal antibodies against undifferentiated cells within the crypt will be generated to expand the available resources for effective isolation of intestinal progenitor and stem cells. The ability to isolate discrete pools of intestinal stem cells is critical for advancing our understanding of intestinal biology, which is paramount for understanding, classifying and treating intestinal disease. PUBLIC HEALTH RELEVANCE: The key to unlocking intestinal biology and harnessing this knowledge for treatment of intestinal disease lies in the ability to identify and isolate the adult intestinal stem cell. This project, designed to understand how this cell functions, will provide insight into diseases where these stem cells do not function properly, such as in intestinal cancer. These studies have the potential for improving diagnoses, informing directed treatment, and designing new therapies for intestinal disease.