Regulation of actively proliferating and quiescent intestinal stem cells

活跃增殖和静止肠道干细胞的调节

• 批准号: 8917929
• 负责人: CALVIN J KUO
• 金额: $ 37.15万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-22 至 2019-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8917929
• 关键词: ALCAM gene; Address; Adenoviruses; Anemia; Back; Bioinformatics; Cell Culture Techniques; Cell division; Cells; Clonality; Collaborations; Colon; Colon Carcinoma; Colorectal Cancer; Columnar Cell; Data; Defect; Development; Duodenum; Epithelial; Exhibits; Frequencies; GRP78 gene; Genetic; Goals; Growth; Health; Hereditary Disease; Homeostasis; Human; In Situ Hybridization; In Vitro; Injury; Intestinal Diseases; Intestines; Iron; Label; Malignant - descriptor; Mediating; Methodology; Methods; Modeling; Molecular; Monitor; National Institute of Diabetes and Digestive and Kidney Diseases; Natural regeneration; Nature; Oncogenic; Organoids; Pathogenesis; Pathway interactions; Phenotype; Population; Positioning Attribute; Proliferating; Reagent; Regulation; Repression; Research Project Grants; SHFM1 gene; Serum; Signal Transduction; Small Intestines; Spatial Distribution; Stem cell transplant; Stem cells; Translations; Transplantation; Up-Regulation; autocrine; base; cancer stem cell; cell behavior; disease phenotype; gain of function; in vivo; loss of function; metal transporting protein 1; microcytic anemia; molecular marker; next generation; novel; novel marker; peripheral blood; progenitor; receptor; reconstitution; regenerative; stem cell biology; stem cell division; stem cell niche; stem cell population; transcriptome sequencing; tumor; villin

DESCRIPTION (provided by applicant): The tremendous regenerative capacity of the intestine, with epithelial turnover every 5-7 days, is mediated by the carefully orchestrated activity of intestinal stem cells (ISC). The recent identification of molecular markers for ISCs, combined with in vivo lineage tracing and in vitro primary organoid culture methodologies has revolutionized ISC biology. The resultant wealth of findings have showcased ISCs as a robust general paradigm for stem cell behavior and have elicited a next generation of studies exploring the ISC niche, interrelationships and interconversion of actively cycling versus quiescent ISC populations, and translational applications of ISC transplantation. Accordingly, the current application is a continuation of our prior 5 year project for the NIDDK Intestinal Stem Cell Consortium (ISCC), and is responsive to RFA-DK-13-012, Intestinal Stem Cell Consortium Research Projects (U01), seeking to "define conditions controlling proliferation and differentiation of intestinal stem cells", "determine the developmental lineage of characterized populations of stem cells in the small intestine" and "methods for expanding and grafting stem cells back into the intestine". Here, we build upon our preliminary data from the prior cycle with novel reagents and methodologies. Aim 1 explores R-spondins as regulators of Lgr5+ ISC symmetric cell division combining adenovirus-mediated gain- and loss-of-function approaches with in vivo lineage tracing, and pursuing mechanistic and niche localization studies. In Aim 2, interrelationships between diverse cycling and quiescent ISC populations will be investigated by RNA-Seq, and novel lineage relationships of Bmi1+ ISC will be explored. Aims 3 and 4 explore translational aspects of ISC biology, with Aim 3 utilizing organoid culture to evaluating the effects of R- spondins on colon cancer stem cells, and Aim 4 attempting the phenotypic correction of a Mendelian intestinal epithelial defect by ISC transplantation. Overall, these studies describe an integrated approach to ISC biology and translation that is highly responsive to RFA-DK-13-012, Intestinal Stem Cell Consortium Research Projects (U01), with implications for stem cell biology, and intestinal disease pathogenesis and therapy.

描述（由申请人提供）：肠道的巨大再生能力，每 5-7 天就有一次上皮更新，是由肠道干细胞 (ISC) 精心策划的活动介导的。最近对 ISC 分子标记的鉴定，结合体内谱系追踪和体外原代类器官培养方法，彻底改变了 ISC 生物学。由此产生的大量研究结果表明，ISC是干细胞行为的一个强大的通用范式，并引发了下一代研究，探索ISC生态位、活跃循环与静止ISC群体的相互关系和相互转换，以及ISC移植的转化应用。因此，当前的申请是我们之前 NIDDK 肠道干细胞联盟 (ISCC) 5 年项目的延续，并响应 RFA-DK-13-012、肠道干细胞联盟研究项目 (U01)，寻求“定义控制肠道干细胞增殖和分化的条件”、“确定小肠干细胞特征群体的发育谱系”以及“扩增和分化的方法”将干细胞移植回肠道”。在这里，我们利用新的试剂和方法建立在前一个周期的初步数据的基础上。目标 1 探索 R-spondins 作为 Lgr5+ ISC 对称细胞分裂的调节剂，将腺病毒介导的功能获得和功能丧失方法与体内谱系追踪相结合，并进行机制和生态位定位研究。在目标 2 中，将通过 RNA-Seq 研究不同循环和静止 ISC 群体之间的相互关系，并将探索 Bmi1+ ISC 的新谱系关系。目标 3 和 4 探索 ISC 生物学的转化方面，目标 3 利用类器官培养来评估 R-spondins 对结肠癌干细胞的影响，目标 4 尝试通过 ISC 移植来纠正孟德尔肠上皮缺陷的表型。总体而言，这些研究描述了一种 ISC 生物学和翻译的综合方法，该方法对 RFA-DK-13-012、肠道干细胞联盟研究项目 (U01) 高度敏感，对干细胞生物学以及肠道疾病发病机制和治疗具有影响。