Regulation of Human Tumorigensis by Cancer Specific NXF1 Adaptor Proteins

癌症特异性 NXF1 接头蛋白对人类肿瘤发生的调节

• 批准号: 10596156
• 负责人: GEORGE L SEN
• 金额: $ 35.42万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-04-01 至 2027-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10596156
• 关键词: 3' Untranslated Regions; 3-Dimensional; Adaptor Signaling Protein; Address; Apoptosis; Applications Grants; Automobile Driving; Basement membrane; Binding; Binding Proteins; Cell Line; Cell Survival; Cells; Code; Complex; Cytoplasm; Data; Dermal; Disease; Epidermis; Family; Gene Expression; Generations; Genes; Genetic Transcription; Growth; Human; Immune; Lead; MAP Kinase Gene; Malignant Neoplasms; Mediating; Melanoma Cell; Messenger RNA; Mitosis; Modeling; Molecular; Mus; Mutation; Natural regeneration; Neoplasms; Normal tissue morphology; Nuclear Pore Complex; Oncogenic; Output; Pathway interactions; Process; Proteins; RNA; Regulation; Reporting; Research Design; Role; Signal Transduction; Skin; Specificity; Squamous cell carcinoma; Transcript; Translating; Tumor Promotion; Tumor stage; VEGFC gene; Vascular Endothelial Growth Factor C; cancer diagnosis; cell motility; clinically relevant; crosslinking and immunoprecipitation sequencing; design; experimental study; human model; in vivo; insight; knock-down; loss of function; mRNA Export; member; neoplastic; new technology; overexpression; pluripotency; programs; protein complex; stem; therapeutic target; therapy development; transcription factor; tumor; tumor initiation; tumor progression; tumorigenesis

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.

项目总结/摘要 背景：癌症起源于基因改变，导致基因突变。 促进肿瘤存活、生长、运动和抑制肿瘤生长的表达程序， 分化和凋亡。这个致癌基因表达程序的mRNA 必须输出到细胞质中翻译成蛋白质，以促进 肿瘤发生是否存在肿瘤特异性mRNA的输出调节， 参与该过程的潜在蛋白质是未知的。我们已经证明肿瘤 特异性NXF 1接头蛋白调节致癌mRNA的输出。适配器 蛋白质在肿瘤起始期间也高度上调， 衔接子抑制肿瘤发生。 目的/假设：本提案旨在了解 使用RAS驱动的人类皮肤来驱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个衔接蛋白是高度 在肿瘤发生过程中上调，与NXF 1相关，介导 致癌基因表达程序。我们的目标是描述 每个肿瘤在正常到肿瘤的进展中诱导NXF 1接头蛋白 皮肤此外，我们试图确定每个衔接蛋白的特定转录本， 在肿瘤起始期间结合以促进肿瘤发生。 具体目的：（1）探讨NXF 1接头蛋白在人肝癌细胞系中的作用 从正常皮肤到肿瘤性皮肤，以及（2）鉴定与NXF 1相关的转录物 衔接蛋白，并确定哪些结合转录本被输出。 研究设计：为了在更临床相关的环境中研究表皮肿瘤发生， 我们生成三维完整的人类皮肤，包含人类表皮细胞（其 已被永久敲除的衔接蛋白）的背景下， 真皮基质和基底膜，在免疫受损小鼠上再生。 通过使用该模型，我们可以在NXF 1适配器上进行功能丧失实验 蛋白质以表征它们在表皮生长中的作用， 分化和进展为瘤形成。我们将使用CLIP-Seq来确定 在从正常到正常的进展过程中，与每个衔接蛋白相关的RNA 肿瘤性表皮