Role of Noncoding RNAs in Epidermal Differentiation

非编码 RNA 在表皮分化中的作用

• 批准号: 8390210
• 负责人: Bryan Sun
• 金额: $ 5.57万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-11-16 至 2015-11-15
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8390210
• 关键词: Beryllium; Binding; Biological Assay; Biology; Cell Cycle Regulation; Cell physiology; Complex; Cytoplasmic Granules; Data; Defect; Down-Regulation; Elements; Epithelial; Foundations; Functional RNA; Future; Genes; Genetic; Genetic Epistasis; Genomics; Goals; Homeostasis; Human Genome; Immune; Individual; Keratin; Knowledge; Laboratories; Literature; Malignant Neoplasms; Mediating; Messenger RNA; Modeling; Molecular; Molecular Mechanisms of Action; Molecular Models; Names; National Research Service Awards; Phenocopy; Process; Protein Microchips; Proteins; RNA; RNA Binding; RNA Interference; RNA-Binding Proteins; RNA-Protein Interaction; Regulation; Role; Series; Skin; Specificity; Stem cells; Surveys; Testing; Tissue Model; Tissues; Transcript; Untranslated RNA; Work; alveolar lamellar body; base; filaggrin; improved; insight; keratinocyte differentiation; loricrin; mRNA Transcript Degradation; molecular modeling; mutant; novel; pluripotency; programs; research study; skin disorder; trafficking

DESCRIPTION (provided by applicant): Long, noncoding RNAs (lncRNAs) have diverse roles in a variety of cellular processes, including stem cell pluripotency, cell cycle control, and immune regulation. LncRNAs function by varying mechanisms, and have the capacity to coordinately regulate the activity of many genes at once. Surveys of the human genome indicate the presence of thousands of lncRNAs. The vast majority have not yet been studied in detail. The goal of this Ruth L. Kirschstein NRSA Postdoctoral Foundation (F32) application is to characterize the role of lncRNAs in epithelial differentiation. Initial work in our laboratory has identified a new LncRNA that is upregulated during keratinocytes differentiation, which has been named terminal differentiation-induced noncoding RNA (TINCR). Depletion of this LncRNA in an epithelial model results in a differentiation defect. RNA-protein interaction assays demonstrate that TINCR interacts with Staufen-1 (Stau1) which is an RNA-binding protein with known roles in RNA trafficking and degradation. Depletion of Stau1 results in an epidermal differentiation defect that phenocopies TINCR depletion. These findings support the potential role of a new LncRNA and its interacting protein in skin differentiation, and provide a framework to further explore their functions in the skin. The first objective of this proposal is to define the functionl domains of TINCR. Using a series of deletion mutants, the function of individual segments of this new LncRNA will be assayed in an organotypic skin model. The second objective of this proposal will be to determine the relationship between TINCR and Stau1. Epistasis experiments will be performed to determine how these elements interact genetically to regulate skin differentiation. Then, specific molecular models of their function will be tested using RNA-interference based genetic "knockdown" experiments. A role for lncRNAs in the homeostasis of somatic tissues, such as skin, has not yet been described in the literature. The lines of experimentation described in this proposal will provide insight to the mechanism of action of a novel LncRNA, TINCR, in the regulation of skin homeostasis and differentiation. Dysregulated skin differentiation is a hallmark of several skin diseases, and improving the knowledge of the molecular regulators of the differentiation process will facilitate approaches to understand the cause and potential treatment of these conditions. PUBLIC HEALTH RELEVANCE: This proposal aims to further characterize the role of long, non-coding RNAs in epidermal homeostasis and differentiation. Defects in epidermal differentiation are a hallmark of several skin diseases, and improving our understanding of the molecular regulators of skin differentiation can provide insight into the cause and potential treatment for these conditions.

描述（由申请人提供）：长非编码 RNA (lncRNA) 在多种细胞过程中具有多种作用，包括干细胞多能性、细胞周期控制和免疫调节。 LncRNA 通过不同的机制发挥作用，并且能够同时协调调节许多基因的活性。对人类基因组的调查表明存在数千个 lncRNA。绝大多数尚未得到详细研究。 Ruth L. Kirschstein NRSA 博士后基金会 (F32) 申请的目标是表征 lncRNA 在上皮分化中的作用。我们实验室的初步工作发现了一种在角质形成细胞分化过程中上调的新 LncRNA，将其命名为终末分化诱导非编码 RNA (TINCR)。上皮模型中该 LncRNA 的缺失会导致分化缺陷。 RNA-蛋白质相互作用测定表明，TINCR 与 Staufen-1 (Stau1) 相互作用，Staufen-1 是一种 RNA 结合蛋白，在 RNA 运输和降解中具有已知的作用。 Stau1 的耗竭会导致表皮分化缺陷，其表型与 TINCR 耗竭相似。这些发现支持了新的 LncRNA 及其相互作用蛋白在皮肤分化中的潜在作用，并为进一步探索它们在皮肤中的功能提供了框架。该提案的第一个目标是定义 TINCR 的功能域。使用一系列缺失突变体，这种新 LncRNA 各个片段的功能将在器官型皮肤模型中进行分析。该提案的第二个目标是确定 TINCR 和 Stau1 之间的关系。将进行上位实验以确定这些元素如何通过遗传相互作用来调节皮肤分化。然后，将使用基于 RNA 干扰的遗传“击倒”实验来测试其功能的特定分子模型。文献中尚未描述 lncRNA 在体细胞组织（例如皮肤）稳态中的作用。该提案中描述的实验路线将深入了解新型 LncRNA TINCR 在调节皮肤稳态和分化方面的作用机制。皮肤分化失调是多种皮肤病的标志，提高对分化过程分子调节因子的了解将有助于了解这些疾病的原因和潜在治疗方法。 公共健康相关性：该提案旨在进一步表征长非编码 RNA 在表皮稳态和分化中的作用。表皮分化缺陷是多种皮肤病的标志，提高我们对皮肤分化分子调节因子的了解可以深入了解这些疾病的原因和潜在的治疗方法。