Role and mechanisms of IGF2BP1 in melanoma pathogenesis

IGF2BP1在黑色素瘤发病机制中的作用和机制

• 批准号: 10304172
• 负责人: Vladimir S. Spiegelman
• 金额: $ 35.49万
• 依托单位: PENNSYLVANIA STATE UNIV HERSHEY MED CTR
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-12-02 至 2024-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10304172
• 关键词: Affect; American Cancer Society; Bioinformatics; Biology; Cell Survival; Cells; Cessation of life; Clinical; Complementary DNA; Cytoplasm; Data; Databases; Development; Diagnosis; Dimensions; Disease-Free Survival; Down-Regulation; Flowcharts; Gene Expression; Genes; Genetic Transcription; Genetically Engineered Mouse; Half-Life; Human; Hypoxia; In Vitro; Malignant Neoplasms; Mediating; Melanoma Cell; Messenger RNA; Metastatic Melanoma; Molecular and Cellular Biology; Neoplasm Metastasis; Outcome; Pathogenesis; Pathway interactions; Patients; Play; Prevention; Primary Neoplasm; Proteins; RNA; RNA-Binding Proteins; Regulation; Resistance; Role; Serum; Testing; The Cancer Genome Atlas; Transcript; Transfer RNA; Translations; Validation; base; cell growth; design; extracellular vesicles; in vivo; insight; mRNA Expression; melanoma; mouse model; new therapeutic target; novel therapeutics; overexpression; technology development; tumor progression; uptake

Melanoma is one of the most lethal forms of cancers. The major reason for lack of satisfactory management of melanomas is our poor understanding of the biology of melanoma pathogenesis. Recently, regulation of mRNA fate in the cytoplasm emerged as a major mechanism controlling gene expression. RNA-binding proteins afford another dimension of the regulation of pathways involved in tumor progression. IGF2BP1 is an RNA-binding protein that regulates stability, localization and translation of its mRNA targets. We have previously shown that RNA-binding protein IGF2BP1 is overexpressed in melanomas, however the role of IGF2BP1 in pathogenesis of melanoma has not been elucidated. Our new preliminary data in different mouse models show that IGF2BP1 affects melanoma metastasis and its high expression in human melanoma correlates with poor clinical outcomes. Based on our previous studies and new exciting observations we hypothesize that IGF2BP1 plays a key role in melanoma progression and metastasis. To test this hypothesis, we propose the following independent, but highly interrelated specific aims: Specific Aim 1. To analyze mechanisms of IGF2BP1 function in melanoma metastasis. This aim is designed to identify IGF2BP1 targets responsible for the function of IGF2BP1 in melanoma metastasis. To this end we propose several complementary approaches to perform an unbiased analysis of IGF2BP1 involvement in melanoma metastasis. Our approach will include: i) analysis or RNA downregulated upon inhibition of IGF2BP1 function in melanoma in vivo; ii) in vivo analysis of cDNAs capable of overcoming the inhibition of melanoma metastases caused by IGF2BP1 down-regulation; iii) analysis of changes in half-life and expression of mRNA upon IGF2BP1 inhibition in melanoma; and iv) identification of mRNAs directly interacting with IGF2BP1 in metastatic melanoma cells. Specific Aim 2. To analyze the role of IGF2BP1 in EV-mediated melanoma progression. This aim is designed to comprehensively evaluate the function of IGF2BP1 in melanoma EV, especially as it relates to their role in tumor progression. Our analysis will include the elucidation of the IGF2BP1 in EV-mediated pre-metastatic niche formation; its role in EV uptake by recipient cells in vitro and in vivo; efficiency of RNA transfer to recipient cells; analysis of EV number, size, composition and cargo; and validation of our findings in EV isolated from the serum of melanoma patients. Impact: On successful completion of this study, we will: (a) understand the contribution and mechanisms of IGF2BP1 involvement in melanoma progression; (b) better understand how IGF2BP1 and its target genes are involved in melanoma metastasis; and (c) establish IGF2BP1 as a novel therapeutic target for melanoma patients. The proposed studies which combine cellular and molecular biology, bioinformatics, technology development, and cutting-edge in vivo experimentation will provide insights into the interplay between the IGF2BP1 and its target molecules and how these contribute to melanoma progression. Ultimately, the data obtained in this study have a potential to pave the way for new therapeutics in melanoma.

黑色素瘤是最致命的癌症形式之一。缺乏令人满意管理的主要原因 黑色素瘤是我们对黑色素瘤发病机理生物学的不良理解。最近，mRNA的调节 细胞质中的命运是控制基因表达的主要机制。 RNA结合蛋白负担得起 涉及肿瘤进展的途径调节的另一个维度。 IGF2BP1是RNA结合 调节其mRNA靶标的稳定性，定位和翻译的蛋白质。我们以前已经表明 RNA结合蛋白IGF2BP1在黑色素瘤中过表达，但是IGF2BP1在发病机理中的作用 黑色素瘤尚未阐明。我们在不同鼠标模型中的新初步数据表明IGF2BP1 影响黑色素瘤转移及其在人黑色素瘤中的高表达与临床结局不良相关。 根据我们以前的研究和新的令人兴奋的观察，我们假设IGF2BP1在 黑色素瘤的进展和转移。为了检验这一假设，我们提出以下独立但高度提议 相互关联的特定目的：特定目的1。分析黑色素瘤中IGF2BP1功能的机制 转移。该目标旨在确定负责IGF2BP1功能的IGF2BP1目标 黑色素瘤转移。为此，我们提出了几种互补的方法来执行公正 IGF2BP1参与黑色素瘤转移的分析。我们的方法包括：i）分析或RNA 在体内黑色素瘤中抑制IGF2BP1功能后，下调； ii）能够的cDNA的体内分析 克服由IGF2BP1下调引起的黑色素瘤转移抑制； iii）分析变化 在黑色素瘤中IGF2BP1抑制时，在半衰期和mRNA表达中；和iv）识别mRNA 在转移性黑色素瘤细胞中与IGF2BP1直接相互作用。特定目的2。分析IGF2BP1的作用 在EV介导的黑色素瘤进展中。此目标旨在全面评估 黑色素瘤EV中的IGF2BP1，尤其是与它们在肿瘤进展中的作用有关。我们的分析将包括 在EV介导的中替代前生态位形成中阐明IGF2BP1；收件人在电动汽车吸收中的作用 细胞体外和体内； RNA转移到受体细胞的效率； EV数量，大小，组成的分析 和货物；以及我们从黑色素瘤患者血清中分离出的EV中发现的结果。影响：ON 成功完成这项研究，我们将：（a）了解IGF2BP1的贡献和机制 参与黑色素瘤进展； （b）更好地了解IGF2BP1及其靶基因如何参与 黑色素瘤转移； （c）将IGF2BP1建立为黑色素瘤患者的新型治疗靶点。这 提出的研究结合了细胞和分子生物学，生物信息学，技术发展和 尖端的体内实验将为IGF2BP1及其目标之间的相互作用提供见解 分子以及它们如何促进黑色素瘤进展。最终，本研究中获得的数据具有 有潜力为黑色素瘤新疗法铺平道路。