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）：已经假设存在真正静止且长寿的干细胞，但迄今为止尚未在肠道中发现。双皮质素样激酶 1 (Dclk1) 被提议作为肠道干细胞标记物，但也发现在肠道簇细胞中表达。 最近，我们的团队培育出了 Dclk1-Cre-ERT BAC 转基因小鼠，并且明确表明 Dclk1+ 簇细胞是长寿的静止干细胞，源自 Lgr5+ 干细胞，但由神经支配诱导。我们的假设是，Dclk1+ 簇细胞既作为肠道干细胞生态位的一部分，又作为储备肠道干细胞，并且是通过外在神经支配产生的。我们将通过三个具体目标来探索这一假设。 (1)神经在Dclk1+簇绒细胞的诱导中起什么作用。我们将利用体外培养系统和体内小鼠模型来检查神经支配的作用，重点关注 NGF-Trk 通路在介导神经簇细胞相互作用中的作用。 (2) Dclk1+祖细胞的消融是否会抑制正常肠上皮稳态和生长以及对辐射损伤的反应？与 DTR F/F 小鼠杂交的 Dclk1-Cre-ERT 小鼠将接受白喉毒素治疗，并使用体外和体内系统评估对辐射的再生反应。 （3）。 Dclk1+ 祖细胞的激活是否会导致辐射损伤后肠道增殖和再生增加？初步研究表明，删除 Dclk1 谱系中的 Apc 并激活 Wnt 信号传导足以将 Dclk1 簇细胞转化为活性干细胞。我们将探讨 Wnt 信号传导的作用 隐窝培养物和小鼠模型。总的来说，这些研究应该为 Dclk1 祖细胞作为肠道中的生态位细胞和干细胞的作用及其贡献提供新的见解 到肠道再生。