Quiescent Dclk1+ stem cells in the mouse intestine

小鼠肠道中的静态 Dclk1 干细胞

基本信息

批准号：
8547458
负责人：
Timothy Cragin Wang
金额：
$ 34.78万
依托单位：
COLUMBIA UNIVERSITY HEALTH SCIENCES
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-09-25 至 2013-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8547458
关键词：
Ablation Adult Behavior Cell Count Cell Lineage Cells Cloning Columnar Cell Data Diphtheria Toxin Embryo Endoderm Enteral Enteric Nervous System Epithelial Epithelial Cells Gastrointestinal tract structure Gene Expression Generations Genes Genetic Growth Homeostasis Image In Vitro Injury Intestines Knock-in Mouse Label Laboratories Life Maintenance Modeling Molecular Profiling Mus Natural regeneration Nerve Nervous system structure Neural Crest Organ PTEN gene Phosphotransferases Play Positioning Attribute Radiation Radiation Injuries Regulation Reserve Stem Cell Role Secretory Cell Signal Transduction Stem cells Stromal Cells System Testing Time Tissues Toxin Transgenic Mice Transgenic Organisms Tumor Suppressor Proteins base bone cell type doublecortin protein insight mouse model neglect nerve stem cell notch protein progenitor regenerative response stem stem cell niche stem cell population

项目摘要

DESCRIPTION (provided by applicant): Truly quiescent and long-lived stem cells have been postulated but to date not identified in the gut. Doublecortin-like kinase 1 (Dclk1) was proposed as a stem cell marker in the intestine but was also found to be expressed in intestinal tuft cells. Recently, our group has generated Dclk1- Cre-ERT BAC transgenic mice, and we have shown unequivocally that Dclk1+ tuft cells are long-lived, quiescent stem cells that originate from the neural crest, and that Dclk1 also labels extraganglionic progenitors of the enteric nervous system. Our hypothesis is that Dclk1+ stem cells play a role both as neural stem cells and also as reserve intestinal stem cells, which together form a niche for other stem cell populations in the intestine, primarily through generation of the ENS. We will explore this hypothesis through four specific aims. (1) What is the relationship between Dclk1+ stem cells and Bmi1+ intestinal stem cells? We will utilize Bmi1/GFP/+ knockin mice and Lgr5/DTR knockin mice in combination with Dclk1 transgenic lines to determine whether there is overlap between Dclk1 and Bmi1. (2) Does ablation of Dclk1+ progenitors inhibit normal intestinal epithelial homeostasis and the response to radiation injury? Dclk1-Cre-ERT mice crossed to DTR F/F mice will be treated with diptheria toxin and the regenerative response to radiation assessed. (3). Does activation of Dclk1+ progenitors result in intestinal proliferation and increased regeneration following radiation injury? Dclk1 progenitors will be activated by conditional deletion of PTEN or activation of IC-Notch1, followed by analysis of the response to radiation injury. (4). Does ablation of Dclk1 progenitors inhibit in vitro ISC growth, and does activation of Dclk1+ progenitors accelerate in vitro ISC growth. We will use whole intestinal cultures derived from the Dclk1 crosses above to explore the effect of Dclk1 ablation or activation on in vitro growth. Overall, these studies should provide new insights into the role of Dclk1 progenitors as both niche and stem cells in the intestine, and their contribution to intestinal regeneration.

描述（由申请人提供）：已经假定了真正的静止和长寿命的干细胞，但迄今为止尚未在肠道中识别。提出了二核素样激酶1（DCLK1）作为肠道中的干细胞标记，但也发现在肠道簇细胞中表达。最近，我们的组产生了DCLK1-CRE-ERT BAC转基因小鼠，我们明确地表明DCLK1+簇细胞是长期寿命的，顽强的干细胞，源自神经rest，DCLK1还标记了肠道神经系统的外离子祖细胞。我们的假设是DCLK1+干细胞既起着神经干细胞的作用，又是储备肠道干细胞，它们共同形成了肠中其他干细胞种群的利基，主要是通过ENS的产生。我们将通过四个特定目标探讨这一假设。（1）DCLK1+干细胞与BMI1+肠干细胞之间的关系是什么？我们将利用BMI1/GFP/+敲击蛋白小鼠和LGR5/DTR敲击蛋白与DCLK1转基因系组合来确定DCLK1和BMI1之间是否存在重叠。（2）DCLK1+祖细胞的消融是否会抑制正常的肠上皮稳态和对辐射损伤的反应？与DTR F/F小鼠交叉的DCLK1-CRE-CRE-ERT小鼠将用白喉毒素治疗，并评估了对辐射的再生反应。（3）。 DCLK1+祖细胞的激活会导致肠道增殖和辐射损伤后再生的增加吗？ DCLK1祖细胞将通过有条件的PTEN或激活来激活 IC-NOTCH1的作品，然后分析对辐射损伤的反应。（4）。 DCLK1的消融祖细胞抑制体外ISC的生长，并且DCLK1+祖细胞的激活加速了体外ISC生长。我们将使用从上面的DCLK1杂交中得出的全肠培养物来探索DCLK1消融或激活对体外生长的影响。总体而言，这些研究应提供有关DCLK1祖细胞在肠道中既有细胞和干细胞的作用及其对肠再生的贡献的新见解。