Cell adhesion and cytoskeletal dynamics in skin

皮肤中的细胞粘附和细胞骨架动力学

• 批准号: 7929089
• 负责人: ELAINE FUCHS
• 金额: $ 25.65万
• 依托单位: ROCKEFELLER UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-22 至 2011-09-21
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7929089
• 关键词: Actins; Address; Adherens Junction; Adhesions; Adhesives; Adult; Binding Proteins; Biochemical; Biology; Cadherins; Cell Adhesion; Cell Lineage; Cells; Cellular biology; Clinical; Cues; Cytoskeletal Gene; Cytoskeletal Proteins; Cytoskeleton; Data; Defect; Development; Diagnosis; Embryo; Embryonic Development; Endoderm Cell; Engineering; Epidermis; Epithelial; Epithelium; Equilibrium; Felis catus; Focal Adhesion Kinase 1; Focal Adhesions; Gene Expression; Genes; Genetic; Global Change; Grant; Growth; Hair; Hair follicle structure; Hereditary Disease; Homeostasis; Human; In Vitro; Integrins; Knowledge; Lead; Life; Link; Malignant Neoplasms; Mediating; Methods; Microfilaments; Microtubules; Molecular; Molecular Genetics; Monitor; Monomeric GTP-Binding Proteins; Morphogenesis; Multipotent Stem Cells; Mus; Phosphotransferases; Physiological; Process; Property; Proteins; Research; Resistance; Role; Signal Transduction; Site; Skin; Skin Cancer; Skin Transplantation; Skin graft; Squamous cell carcinoma; Stem cells; Stratification; Structure; Surface; Testing; Time; Tissues; Transgenes; Viral; Wound Healing; Wounds and Injuries; appendage; base; blastocyst; cell motility; human disease; improved; in utero; in vivo; innovation; insight; keratinocyte; link protein; novel; polymerization; public health relevance; repaired; response; self-renewal; skin disorder; skin morphogenesis; small hairpin RNA; tissue regeneration; tool; wound

DESCRIPTION (provided by applicant): Our global objective is to develop a molecular understanding of how epithelial stem cells undergo morphogenesis, homeostasis, and wound repair in mammalian skin and bring this research to a clinical setting. Our focus is on how skin stem cells utilize cytoskeletal connections to intercellular and cell substratum junctions to generate tissues. Knowledge of the proteins involved in coordinating actin filament (AF) cytoskeletal dynamics with adherens junctions (AJs) and elucidating how these connections are inversely coordinated with integrin-AF dynamics are key to understanding how self-renewing skin epithelium maintains and repairs its surface barrier and how a stem cell can give rise to a hair follicle (HF). Elucidating how microtubules (MTs) link to AJs is important for understanding how the stratified epidermis maintains a single layer of dividing cells, and how spindle polarity changes when stem cells are activated to produce HFs. A molecular understanding of these dynamics will be obtained by focusing on four cytoskeletal-junction linking proteins that surfaced as being key from our prior studies: -catenin, which coordinates and polarizes AF-AJ dynamics and suppresses AF-integrin dynamics; ACF7, which coordinates and polarizes MT-AF and Par3- mediated dynamics in a wound response; and Par3 and mInsc, whose polarization relies upon -catenin and 1 integrin, and which appear to be critical for orienting spindles in skin morphogenesis. Through biochemical and molecular approaches, we'll define and characterize the associated proteins that are involved in polarizing and remodeling cytoskeletal dynamics during skin morphogenesis and wound repair. Through mouse genetics and novel ShRNA strategies we've developed to rapidly knockdown genes in adult and embryonic skin, the functional significance of these proteins and their varied associations with other proteins will be ascertained. Finally, as the major players in these processes unfold, functional analyses will be combined with microarray data to define the underlying significance of global changes in AJ-cytoskeletal gene expression that occur concomitantly as stem cells receive external cues to remodel their AJ-cytoskeletal connections and initiate stratification and HF morphogenesis. Understanding how these cytoskeletal linkages are regulated in normal skin is a prerequisite to elucidating how defects in these processes lead to genetic disorders, including skin cancers. Past and present AR27883 research provides an excellent illustration of how molecular skin biology can help to generate new and improved tools for the diagnosis and treatment of human skin disease. Public Health Relevance: Stem cells are natural units of tissue repair and homeostasis, and their versatility holds promise for tissue regeneration. This research focuses on deciphering how different tissue structures are derived from multipotent stem cells in the skin. Specifically, we focus on how stem cells within a single layer use cell-cell and cell substratum junctions to remodel their cytoskeleton to generate a stratified, differentiating epidermis or an invaginating hair follicle bud, and how this changes transiently in a wound response. This study is a fundamental prerequisite to understanding how aberrations in these basic properties go awry in skin cancers, including squamous cell carcinomas.

描述（由申请人提供）：我们的全球目标是从分子水平了解上皮干细胞如何在哺乳动物皮肤中进行形态发生、稳态和伤口修复，并将这项研究应用于临床。我们的重点是皮肤干细胞如何利用细胞骨架连接到细胞间和细胞基质连接产生组织。了解参与协调肌动蛋白丝（AF）细胞骨架动力学与粘附连接（AJs）的蛋白质，并阐明这些连接如何与整合素AF动力学反向协调，是理解自我更新皮肤上皮细胞如何维持和修复其表面屏障以及干细胞如何产生毛囊（HF）的关键。阐明微管（MT）如何连接到AJs对于理解分层表皮如何保持单层分裂细胞以及当干细胞被激活产生HF时纺锤体极性如何变化非常重要。这些动力学的分子理解将通过集中在四个细胞间隙连接蛋白获得，这些蛋白从我们先前的研究中作为关键浮出水面：-连环蛋白，其协调和极化AF-AJ动力学并抑制AF-整联蛋白动力学; ACF 7，其协调和极化创伤反应中的MT-AF和Par 3介导的动力学;以及Par 3和mInsc，其极化依赖于β-连环蛋白和β-整联蛋白，并且似乎对于皮肤形态发生中的纺锤体定向至关重要。通过生物化学和分子方法，我们将定义和表征相关蛋白，这些蛋白在皮肤形态发生和伤口修复过程中参与极化和重塑细胞骨架动力学。通过小鼠遗传学和新的shRNA策略，我们已经开发出快速敲除成人和胚胎皮肤中的基因，这些蛋白质的功能意义及其与其他蛋白质的各种关联将被确定。最后，随着这些过程中的主要参与者的展开，功能分析将与微阵列数据相结合，以确定伴随干细胞接受外部线索以重塑其AJ-cytoskeleton连接并启动分层和HF形态发生的AJ-cytoskeleton基因表达的全球变化的潜在意义。了解这些细胞骨架连接在正常皮肤中是如何调节的，是阐明这些过程中的缺陷如何导致遗传疾病（包括皮肤癌）的先决条件。过去和现在的AR 27883研究提供了一个很好的例子，说明分子皮肤生物学如何帮助产生新的和改进的工具，用于诊断和治疗人类皮肤病。 公共卫生相关性：干细胞是组织修复和体内平衡的天然单位，它们的多功能性为组织再生带来了希望。这项研究的重点是破译不同的组织结构是如何从皮肤中的多能干细胞衍生出来的。具体来说，我们专注于干细胞如何在一个单一的层使用细胞和细胞基质连接重塑他们的细胞骨架，以产生分层，分化表皮或内陷毛囊芽，以及如何在伤口反应中发生短暂的变化。这项研究是了解这些基本性质的畸变如何在皮肤癌（包括鳞状细胞癌）中出错的基本先决条件。