Role of Noncoding RNAs in Epidermal Differentiation

描述（由申请人提供）：长链非编码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。绝大多数尚未得到详细研究。Ruth L. Kirschstein NRSA博士后基金（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新的LncRNA，在角质形成细胞分化过程中上调，被命名为终端分化诱导的非编码RNA （terminal differentiation-induced noncoding RNA， TINCR）。在上皮细胞模型中，这种LncRNA的缺失会导致分化缺陷。RNA-蛋白相互作用分析表明，TINCR与Staufen-1 （Stau1）相互作用，Stau1是一种RNA结合蛋白，已知在RNA运输和降解中起作用。Stau1的缺失导致表皮分化缺陷，表型上反映了TINCR的缺失。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。本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，将在器官型皮肤模型中检测该新LncRNA单个片段的功能。本建议的第二个目标将是确定TINCR与Stau1之间的关系。将进行上位实验，以确定这些元素如何在遗传上相互作用以调节皮肤分化。然后，将使用基于rna干扰的基因“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）稳态中的作用尚未在文献中描述。本提案中描述的实验线将提供对新型LncRNA （TINCR）在调节皮肤稳态和分化中的作用机制的见解。皮肤分化失调是几种皮肤病的标志，提高对分化过程分子调节因子的认识将有助于了解这些疾病的原因和潜在治疗方法。

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.