Novel regulators of skin development and Foxn1 function

皮肤发育和 Foxn1 功能的新型调节因子

• 批准号: 8225398
• 负责人: JANICE L BRISSETTE
• 金额: $ 33.35万
• 依托单位: SUNY DOWNSTATE MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-03-01 至 2014-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8225398
• 关键词: Animals; Biochemical; Biological Assay; Biological Models; Cells; Charge; Complement; Complex; Congenital Alopecia; Cultured Cells; Cutaneous; DNA Binding Domain; Defect; Development; Developmental Process; Differentiation and Growth; Disease; Dissection; Environment; Epidermis; Epithelial; Epithelial Cells; Epithelium; Exhibits; Family; Gap Junctions; Gene Expression; Genetic; Genetic Screening; Goals; Growth; Growth Factor; Hair; Hair follicle structure; Hazardous Chemicals; Human; Individual; Link; Mammary gland; Mature T-Lymphocyte; Methods; Modeling; Molecular; Morphogenesis; Mus; Mutation; Nail plate; Natural regeneration; Nature; Organ; Orthologous Gene; Other Genetics; Output; Pathology; Pathway interactions; Pattern; Phenotype; Physiological; Pigmentation physiologic function; Pigments; Play; Process; Production; Property; Proteins; Recruitment Activity; Research; Rodent; Role; Screening procedure; Side; Signal Transduction; Site; Skin; Skin Pigmentation; Skin part; Structure; System; T-Cell Immunodeficiency; Testing; Thymus Gland; Transactivation; Transcription Coactivator; Ultraviolet Rays; Water; Winged Helix; Work; base; cell type; genetic manipulation; insight; keratinocyte; melanocyte; member; novel; pathogen; programs; protein complex; repaired; research study; self-renewal; skin morphogenesis; skin regeneration; trait; transcription factor

DESCRIPTION (provided by applicant): The goal of this project is to identify the molecular partners of Foxn1 and to elucidate their contributions to skin development. Foxn1 is a transcription factor containing a winged-helix DNA-binding domain and a negatively charged transactivation domain. In rodents, the loss of Foxn1 function results in the nude phenotype, which is characterized by the abnormal morphogenesis of the skin, thymus, and mammary gland. To promote skin development, Foxn1 performs an unusual function, which was delineated by us in previous studies. Foxn1 becomes activated as epithelial cells initiate terminal differentiation in the epidermis and hair follicles. In a site-dependent manner, Foxn1 then plays up to three roles: 1) it allows its host cell to differentiate properly, 2) it stimulates the host cell's neighbors to divide, and 3) it recruits melanocytes to the host cell, thus identifying its host as a target for pigmentation. Through this combination of actions, Foxn1 enables groups of cells to work in concert and drives epithelial growth, differentiation, and pigmentation forward together. Presumably, Foxn1 itself works in concert with other factors, which provide it with information, determine its level of activity, direct it to specific targets, or function side-by-side with it in synergistic fashion. Undoubtedly, Foxn1 requires these partners for efficacy, making the partners of Foxn1 essential to the morphogenesis of the skin and other organs. During the course of this project, we will identify the partners of Foxn1 using two approaches, one genetic, the other biochemical. In the genetic approach, we will screen for mutations that modulate the activity or output of Foxn1. The screen will employ a novel model system, which we have developed for the study. In the biochemical approach, we will purify Foxn1 protein complexes from keratinocytes and dissect the individual components. Once partners are identified, we will determine how they contribute to the actions of Foxn1 and the morphogenesis of the skin. In humans, FOXN1 is conserved in sequence and function, suggesting a like conservation of partners. Accordingly, by elucidating the partnerships of Foxn1, the project should provide insight into the development of normal and diseased human skin, most especially, the disorders marked by the aberrant proliferation, differentiation, or pigmentation of epithelial cells. Project Narrative: This project will elucidate fundamental mechanisms by which the skin develops and regenerates its protective epithelial traits. Specifically, the work will delineate a genetic network that drives and coordinates the growth, differentiation, and pigmentation of the skin's external structures. In the short term, the project will explain in part how the skin produces and assembles its barrier to the environment, which provides essential protection against pathogens, hazardous chemicals, ultraviolet light, and water loss. Over the long term, the project will provide insight into how skin may be clinically generated or manipulated, thus facilitating methods for the replacement or repair of damaged and diseased skin.

描述（由申请人提供）：该项目的目标是确定 Foxn1 的分子伙伴并阐明它们对皮肤发育的贡献。 Foxn1 是一种转录因子，含有翼状螺旋 DNA 结合结构域和带负电荷的反式激活结构域。在啮齿类动物中，Foxn1 功能的丧失会导致裸表型，其特征是皮肤、胸腺和乳腺的形态发生异常。为了促进皮肤发育，Foxn1 发挥了一种不寻常的功能，我们在之前的研究中已经描述了这一点。当上皮细胞在表皮和毛囊中启动终末分化时，Foxn1 就会被激活。然后，Foxn1 以位点依赖性方式发挥三种作用：1）它允许其宿主细胞正确分化，2）它刺激宿主细胞的邻居细胞分裂，3）它将黑素细胞招募到宿主细胞，从而识别它的宿主作为色素沉着的目标。通过这种组合作用，Foxn1 使细胞群能够协同工作，并共同推动上皮生长、分化和色素沉着。据推测，Foxn1 本身与其他因素协同作用，为它提供信息，确定其活动水平，将其引导至特定目标，或以协同方式与其并肩发挥作用。毫无疑问，Foxn1 需要这些伙伴才能发挥作用，这使得 Foxn1 的伙伴对于皮肤和其他器官的形态发生至关重要。在这个项目的过程中，我们将使用两种方法来识别 Foxn1 的伙伴，一种是遗传方法，另一种是生物化学方法。在遗传方法中，我们将筛选调节 Foxn1 活性或输出的突变。该屏幕将采用我们为研究开发的新颖模型系统。在生化方法中，我们将从角质形成细胞中纯化 Foxn1 蛋白复合物并解剖各个成分。一旦确定了合作伙伴，我们将确定它们如何促进 Foxn1 的作用和皮肤的形态发生。在人类中，FOXN1 在序列和功能上是保守的，这表明伴侣也有类似的保守性。因此，通过阐明 Foxn1 的伙伴关系，该项目应该可以深入了解正常和患病人类皮肤的发育，尤其是以上皮细胞异常增殖、分化或色素沉着为标志的疾病。项目叙述：该项目将阐明皮肤发育和再生其保护性上皮特征的基本机制。具体来说，这项工作将描绘一个驱动和协调皮肤外部结构的生长、分化和色素沉着的遗传网络。在短期内，该项目将部分解释皮肤如何产生和组装其对环境的屏障，这提供了针对病原体、危险化学品、紫外线和水分流失的基本保护。从长远来看，该项目将深入了解如何在临床上生成或操纵皮肤，从而促进更换或修复受损和患病皮肤的方法。