Quiescent Dclk1+ stem cells in the mouse intestine

小鼠肠道中的静态 Dclk1 干细胞

• 批准号: 8577370
• 负责人: Timothy Cragin Wang
• 金额: $ 32万
• 依托单位: COLUMBIA UNIVERSITY HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-09-15 至 2016-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8577370
• 关键词: Ablation; Adult; Cell Communication; Cell Count; Cell Differentiation process; Cells; Cilia; Coculture Techniques; Data; Development; Diphtheria Toxin; Electron Microscopy; Embryo; Enteral; Enteric Nervous System; Epithelial; Exhibits; Generations; Genes; Growth; Homeostasis; Human; In Vitro; Inflammation; Injury; Intestines; Label; Laboratories; Life; Longevity; Mediating; Modeling; Mus; Natural regeneration; Nerve; Neural Crest; Neurites; Neuroglia; Neurons; Organ; Organoids; Pathway interactions; Phosphotransferases; Play; Property; Radial; Radiation; Radiation Injuries; Regulation; Reporting; Reserve Stem Cell; Role; Signal Transduction; Stem cells; Stromal Cells; Suggestion; Synapses; System; Testing; Tissues; Toxin; Transgenic Mice; beta catenin; cell type; density; doublecortin protein; gastrointestinal; gastrointestinal transplantation; in vivo; inhibitor/antagonist; insight; intestinal crypt; intestinal homeostasis; mouse model; nerve supply; progenitor; public health relevance; regenerative; relating to nervous system; response; sensor; stem; stem cell niche

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 are derived from Lgr5+ stem cells but induced by nervous innervation. Our hypothesis is that Dclk1+ tuft cells play a role both as part of the intestinal stem cell niche and also as reserve intestinal stem cell, and are generated through extrinsic neural innervation. We will explore this hypothesis through three specific aims. (1) What is the role of nerves in the induction of Dclk1+ tuft cells. We will utilize both in vitro culture systems, and in vivo mouse models, to examine the role of nervous innervation, with a focus on NGF-Trk pathway in mediating the nerve-tuft cell interactions. (2) Does ablation of Dclk1+ progenitors inhibit normal intestinal epithelial homeostasis and growth, 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 using both in vitro and in vivo systems. (3). Does activation of Dclk1+ progenitors result in intestinal proliferation and increased regeneration following radiation injury? Preliminary studies suggestion that deletion of Apc in the Dclk1 lineage, and activation of Wnt signaling, is sufficien to convert Dclk1 tuft cells into active stem cells. We will explore the role of Wnt signaling using both crypt cultures and mouse models. 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）：真正的静止和长寿的干细胞已经被假定，但迄今为止尚未确定在肠道。双皮质素样激酶1（Dclk 1）被认为是肠中的干细胞标志物，但也发现在肠簇细胞中表达。 最近，我们的小组已经产生了Dclk 1- Cre-ERT BAC转基因小鼠，我们已经明确表明，Dclk 1+簇细胞是长寿命的，静止的干细胞，来源于Lgr 5+干细胞，但由神经支配诱导。我们的假设是，Dclk 1+簇细胞发挥的作用，既作为肠道干细胞龛的一部分，也作为储备肠道干细胞，并通过外部神经支配产生。我们将通过三个具体目标来探讨这一假设。(1)神经在诱导Dclk 1+簇细胞中的作用是什么？我们将利用体外培养系统和体内小鼠模型来研究神经支配的作用，重点是NGF-Trk通路在介导神经丛细胞相互作用中的作用。(2)消融Dclk 1+祖细胞是否会抑制正常肠上皮细胞的稳态和生长以及对辐射损伤的反应？将用白喉毒素处理与DTR F/F小鼠杂交的Dclk 1-Cre-ERT小鼠，并使用体外和体内系统评估对辐射的再生反应。（三）、放射损伤后Dclk 1+祖细胞的激活是否导致肠增殖和再生增加？初步研究表明，Dclk 1谱系中Apc的缺失和Wnt信号转导的激活有助于将Dclk 1簇细胞转化为活性干细胞。我们将探索Wnt信号转导的作用， 包括隐窝培养物和小鼠模型。总的来说，这些研究应该为Dclk 1祖细胞作为肠道中的小生境和干细胞的作用及其对肠道疾病的贡献提供新的见解。 肠道再生