Genetic Analysis of MicroRNA Functions in Skin Stem Cells In Vivo

体内皮肤干细胞 MicroRNA 功能的遗传分析

• 批准号: 10544018
• 负责人: Rui Yi
• 金额: $ 46.07万
• 依托单位: NORTHWESTERN UNIVERSITY AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-07-01 至 2024-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10544018
• 关键词: Actins; Address; Adherens Junction; Adult; Affect; Animals; Basal Cell; Biological Process; Biology; Cell Adhesion; Cell Migration Induction; Cell Proliferation; Cell division; Cell membrane; Cells; Computer Analysis; Cytoskeleton; Data; Embryo; Environmental Hazards; Epidermis; Epithelial Cells; Equilibrium; Gene Expression; Gene Expression Regulation; Genes; Genetic; Genomics; Germ; Growth; Hair; Hair follicle structure; Homeostasis; Image; Individual; Ion Channel; Knowledge; Liquid substance; Longitudinal Studies; Maintenance; Maps; Mechanics; Mediating; Messenger RNA; MicroRNAs; Molecular; Molecular Probes; Mus; Organ; Pathogenesis; Pathway interactions; Piezo 1 ion channel; Play; Proliferating; Property; RNA; Regenerative Medicine; Regulation; Role; Site; Skin; Skin Aging; Stratum Basale; Stress; Techniques; Therapeutic; Tissues; Untranslated RNA; Visualization; aged; appendage; cell motility; cell type; cellular imaging; computational pipelines; epithelial stem cell; experimental analysis; experimental study; genetic analysis; genomic tools; imaging capabilities; imaging system; in vivo; insight; live cell imaging; mechanotransduction; migration; mouse genetics; mouse model; multi-photon; novel; posttranscriptional; premature; skin organogenesis; stem; stem cell migration; stem cell population; stem cells; tool; transcriptome

Project Summary: Mammalian skin and its appendages function as the outermost barrier of the body to protect inner organs from environmental hazards and keep essential fluids within the body. Homeostasis and integrity of mammalian skin are maintained by multiple progenitor and stem cell populations residing in distinct skin compartments such as basal cells in the interfollicular epidermis and bulge stem cells in hair follicles. In epithelial cells, cell adhesion, migration and proliferation are fundamental properties that are controlled by many mechanisms. Among key regulators, microRNAs (miRNAs) are a class of small, noncoding RNAs that take essential roles in mammalian gene regulation in diverse cell types and tissues. Despite modest regulation of individual targets, miRNAs broadly modulate a large number (60%) of genes and play important roles in a wide range of biological processes. In mammalian skin, the critical functions of the entire miRNA pathway in both embryonic skin development and maintenance of adult HF lineages have been well appreciated. In contrast, the knowledge of individual miRNAs for their targets and function remains scarce. Importantly, similar to other regulators, how miRNAs regulate cell migration and proliferation has not been examined in the context of intact skin in live animals. To address these important issues, we have developed techniques to directly capture miRNA and their targeted mRNA fragments and to image cell migration and proliferation as well as cytoskeleton dynamics in intact skin of live animals. Using these state-of-art tools together with our mouse models, we find that miR-205, the most highly expressed miRNA in epithelial stem cells, promotes cell migration by targeting components of adherens junctions, actin cytoskeleton and mechanosensing genes in both epidermis and hair follicles. In this project, we will further examine how miR-205-regulated cell migration alters the balance between epidermal proliferation and differentiation in the epidermis (Aim 1); how miR-205- induced cell migration triggers hair follicle growth in young and aged mice and how enhanced hair follicle growth affects hair follicle stem cells (Aim 2); and probe how the loss of Piezo1, a mechanically activated ion channel and a new miR-205 target, governs the quiescence of hair follicle stem/progenitor cells (Aim 3). Taken together, studies proposed here, if successful, will significantly enhance our knowledge about mechanisms mediated by individual miRNAs that govern cell migration and proliferation in live animals. The knowledge gained from these studies under normal and stressed conditions will pave the way to manipulate miRNAs and utilize epithelial stem cells for regenerative medicine.

项目概要： 哺乳动物的皮肤及其附属器作为身体最外层的屏障来保护 内部器官免受环境危害，并保持体内必需的液体。 哺乳动物皮肤的稳态和完整性由多个祖细胞和干细胞维持 细胞群存在于不同的皮肤区室中，例如毛囊间的基底细胞 表皮和毛囊中的凸起干细胞。在上皮细胞中，细胞粘附、迁移和 增殖是受多种机制控制的基本特性。其中关键 microRNA (miRNA) 是一类发挥重要作用的小型非编码 RNA 哺乳动物不同细胞类型和组织中的基因调控。尽管监管力度不大 单个目标，miRNA 广泛调节大量 (60%) 基因并发挥重要作用 在广泛的生物过程中发挥作用。在哺乳动物皮肤中， 胚胎皮肤发育和成人 HF 维持中的整个 miRNA 通路 血统得到了很好的赞赏。相反，对单个 miRNA 的了解 目标和功能仍然稀缺。重要的是，与其他调节因子类似，miRNA 如何 调节细胞迁移和增殖尚未在完整皮肤的背景下进行过检查 活的动物。为了解决这些重要问题，我们开发了直接 捕获 miRNA 及其目标 mRNA 片段并对细胞迁移和增殖进行成像 以及活体动物完整皮肤的细胞骨架动力学。使用这些最先进的工具 与我们的小鼠模型一起，我们发现 miR-205，在小鼠中表达最高的 miRNA 上皮干细胞，通过靶向粘附连接的成分促进细胞迁移， 表皮和毛囊中的肌动蛋白细胞骨架和机械感应基因。在这个 项目中，我们将进一步研究 miR-205 调节的细胞迁移如何改变平衡 表皮增殖和表皮分化之间的关系（目标 1）； miR-205-如何 诱导细胞迁移触发年轻和老年小鼠的毛囊生长以及如何增强 毛囊生长影响毛囊干细胞（目标 2）；并探究 Piezo1 的损耗如何 机械激活的离子通道和新的 miR-205 靶点控制头发的静止 卵泡干/祖细胞（目标 3）。综上所述，这里提出的研究如果成功的话，将 显着增强我们对个体 miRNA 介导机制的了解 控制活体动物中的细胞迁移和增殖。从这些研究中获得的知识 在正常和应激条件下将为操纵 miRNA 和利用 miRNA 铺平道路 用于再生医学的上皮干细胞。