Role of Noncoding RNAs in Epidermal Differentiation

描述(由申请人提供)：长的非编码RNA(LncRNAs)在各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。LncRNAs通过不同的机制发挥作用，并具有同时协调调节多个基因活性的能力。对人类基因组的调查表明，存在数以千计的lncRNA。绝大多数还没有被详细研究过。这项Ruth L.Kirschstein NRSA博士后基金会(F32)申请的目标是表征lncRNAs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的LncRNA，被命名为末端分化诱导的非编码RNA(TINCR)。在上皮模型中，这种lncRNA的耗尽会导致分化缺陷。RNA-蛋白质相互作用分析表明，TINCR与Staufen-1(Stau1)相互作用，Staufen-1是一种RNA结合蛋白，已知在RNA运输和降解中发挥作用。Stau1的缺失会导致表皮分化缺陷，从而导致TINCR的缺失。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一个框架。这项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，这种新的lncRNA的单个片段的功能将在器官型皮肤模型中进行测试。这项提案的第二个目标将是确定TINCR和STATU1之间的关系。将进行上位性实验，以确定这些元素如何在遗传上相互作用来调节皮肤分化。然后，将使用基于RNA干扰的遗传“敲除”实验来测试它们功能的特定分子模型。InncRNAs在体细胞组织(如皮肤)的动态平衡中的作用尚未在文献中描述。这项建议中描述的实验路线将为深入了解一种新型的lncRNA，TINCR，在调节皮肤动态平衡和分化方面的作用机制提供帮助。皮肤分化异常是几种皮肤病的一个特征，提高对分化过程的分子调节机制的了解将有助于了解这些疾病的原因和潜在的治疗方法。 公共卫生相关性：这项建议旨在进一步表征长的、非编码的RNA在表皮动态平衡和分化中的作用。表皮分化缺陷是几种皮肤病的标志，提高我们对皮肤分化的分子调节机制的了解可以深入了解这些疾病的原因和潜在的治疗方法。

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.