Regulation of Human Tumorigensis by Cancer Specific NXF1 Adaptor Proteins

项目摘要/摘要 背景：癌症起源于导致基因改变的基因改变 促进肿瘤存活、生长、运动和抑制的表达程序 分化和细胞凋亡。来自致癌基因表达程序的mRNAs 必须输出到细胞质才能转化为蛋白质才能促进 肿瘤发生学。是否有对肿瘤特异性mRNAs输出的监管，以及 参与这一过程的潜在蛋白质尚不清楚。我们已经证明了这种肿瘤 特定的Nxf1接头蛋白调节致癌mRNAs的输出。适配器 在肿瘤的启动和特异性基因的敲除过程中，蛋白质也高度上调。 适配器抑制肿瘤的形成。 目标/假设：这个提议试图理解分子机制。 使用RAS驱动的人推动从正常皮肤到肿瘤皮肤的进展 表皮肿瘤模型。我们的初步数据表明，4个接头蛋白是高度 在肿瘤启动过程中上调，与Nxf1相关，介导 致癌基因表达程序。我们的目标是描述 每种肿瘤诱导的Nxf1接头蛋白在正常到肿瘤进展过程中的作用 皮肤。此外，我们试图确定每个接头蛋白的特定转录本 在肿瘤启动过程中结合以促进肿瘤的形成。 具体目标：(1)确定Nxf1接头蛋白在疾病进展中的作用 从正常皮肤到肿瘤皮肤；(2)鉴定与Nxf1相关的转录本 接头蛋白，并决定输出哪些结合的转录本。 研究设计：为了在更具临床相关性的环境中研究表皮肿瘤的发生， 我们产生了三维完好的人类皮肤，其中包含人类表皮细胞(即 已经被永久地敲除以获得适配蛋白) 真皮基质和基底膜，在免疫受损的小鼠身上再生。 利用该模型，我们可以对NxF1适配器进行功能损失实验 再生的人类皮肤中的蛋白质，以表征它们在表皮生长中的作用， 分化，并进展为肿瘤。我们将使用CLIP-SEQ来确定 在从正常到进化过程中与每个接头蛋白相关的RNA 肿瘤性表皮。

Project Summary/Abstract Background: Cancer originates from genetic alterations that lead to changes in gene expression programs that promote tumor survival, growth, motility and inhibits differentiation and apoptosis. The mRNAs from this oncogenic gene expression program must be exported to the cytoplasm to be translated into protein in order to promote tumorigenesis. Whether there is regulation of export of tumor specific mRNAs and the potential proteins involved in this process is unknown. We have shown that tumor specific NXF1 adaptor proteins regulate export of oncogenic mRNAs. The adaptor proteins are also highly upregulated during tumor initiation and knockdown of specific adaptors inhibit tumorigenesis. Objective/hypothesis: This proposal seeks to understand the molecular mechanisms driving the progression from normal to neoplastic skin using a RAS driven human epidermal tumor model. Our preliminary data suggests that 4 adaptor proteins are highly upregulated during tumor initiation that associates with NXF1 to mediate the export of the oncogenic gene expression program. Our objective is to characterize the role of each tumor induced NXF1 adaptor protein in the progression of normal to neoplastic skin. Furthermore we seek to determine the specific transcripts that each adaptor protein binds during tumor initiation to promote tumorigenesis. Specific Aims: (1) To determine the role of NXF1 adaptor proteins in the progression from normal to neoplastic skin and (2) to identify the transcripts associated with NXF1 adaptor proteins and determine which bound transcripts are exported. Study Design: To study epidermal tumorigenesis in a more clinically relevant setting, we generate 3-dimensionally intact human skin, containing human epidermal cells (that have been permanently knocked down for adaptor proteins) in the context of human dermal stroma and basement membrane, regenerated on immune compromised mice. By using this model, we can perform loss of function experiments on NXF1 adaptor proteins in regenerated human skin to characterize their role in epidermal growth, differentiation, and progression to neoplasia. We will use CLIP-Seq to determine the RNAs associated with each adaptor protein during the progression from normal to neoplastic epidermis.