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，我们明确地证明了DCLK1+ tuft细胞是长期寿命的，静止的干细胞，这些干细胞源自LGR5+干细胞，但由神经支配诱导。我们的假设是DCLK1+ Tuft细胞既是肠道干细胞生态位的一部分，又是储备肠干细胞的作用，并且是通过外在神经神经支配而产生的。我们将通过三个具体目标探讨这一假设。 （1）神经在DCLK1+簇细胞诱导中的作用是什么。我们将利用体外培养系统和体内小鼠模型来检查神经神经神经的作用，重点是NGF-TRK途径在介导神经-TUFT细胞相互作用中。 （2）DCLK1+祖细胞的消融是否会抑制正常的肠上皮稳态和生长，以及对辐射损伤的反应？交叉到DTR F/F小鼠的DCLK1-CRE-ERT小鼠将用白喉毒素治疗，并使用体外和体内系统评估对辐射的再生反应。 （3）。 DCLK1+祖细胞的激活会导致肠道增殖和辐射损伤后再生的增加吗？初步研究表明，DCLK1谱系中APC的缺失以及Wnt信号的激活是足够的，可以将DCLK1 Tuft细胞转换为活性干细胞。我们将探索使用Wnt信令的作用 地穴培养物和小鼠模型。总体而言，这些研究应提供有关DCLK1祖细胞在肠道中的细胞和干细胞的作用的新见解，及其贡献 肠再生。