Role of Noncoding RNAs in Epidermal Differentiation

非编码 RNA 在表皮分化中的作用

• 批准号: 8565649
• 负责人: Bryan Sun
• 金额: $ 5.89万
• 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/8565649
• 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.

描述（由申请人提供）：长的非编码RNA（lncRNA）在多种细胞过程中具有不同的作用，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，并且具有同时协调调节许多基因活性的能力。对人类基因组的调查表明存在数千种lncRNA。其中绝大多数尚未得到详细研究。这个Ruth L的目标。Kirschstein NRSA博士后基金会（F32）申请旨在表征lncRNA在上皮分化中的作用。我们实验室的初步工作已经确定了一种新的LncRNA，在角质形成细胞分化过程中上调，被命名为终末分化诱导的非编码RNA（TINCR）。在上皮模型中这种LncRNA的消耗导致分化缺陷。RNA-蛋白质相互作用试验表明，TINCR与Staufen-1（Stau 1）相互作用，Stau 1是一种RNA结合蛋白，在RNA运输和降解中具有已知的作用。Stau 1的消耗导致表皮分化缺陷，其表型模仿TINCR消耗。这些发现支持了一种新的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一个框架。该建议的第一个目标是定义TINCR的功能域。使用一系列缺失突变体，将在器官型皮肤模型中测定这种新LncRNA的各个片段的功能。本提案的第二个目标是确定TINCR和Stau 1之间的关系。上位性实验将进行，以确定这些元素如何相互作用，遗传调节皮肤分化。然后，将使用基于RNA干扰的遗传“敲低”实验来测试其功能的特定分子模型。lncRNA在体细胞组织如皮肤的稳态中的作用尚未在文献中描述。本提案中描述的实验路线将为新型LncRNA TINCR在调节皮肤稳态和分化中的作用机制提供见解。失调的皮肤分化是几种皮肤疾病的标志，提高分化过程的分子调节剂的知识将有助于了解这些疾病的原因和潜在治疗方法。