Role and mechanisms of IGF2BP1 in melanoma pathogenesis

黑色素瘤是最致命的癌症之一。缺乏令人满意的管理的主要原因 黑色素瘤是我们对黑色素瘤发病机制的生物学认识不足。最近，mRNA的调节 细胞质中的命运成为控制基因表达的主要机制。RNA结合蛋白提供 另一个方面的调节途径参与肿瘤的进展。IGF2BP1是一种RNA结合蛋白， 调节其mRNA靶点的稳定性、定位和翻译的蛋白质。我们之前已经证明， RNA结合蛋白IGF2BP1在黑色素瘤中过表达，然而IGF2BP1在发病机制中的作用 黑色素瘤的病因尚未阐明。我们在不同小鼠模型中的新的初步数据表明，IGF2BP 1 影响黑色素瘤转移，并且其在人黑色素瘤中的高表达与不良临床结果相关。 基于我们以前的研究和新的令人兴奋的观察，我们假设IGF2BP1在 黑素瘤进展和转移。为了验证这一假设，我们提出了以下独立的，但高度 相关的具体目标：具体目标1。分析IGF2BP1在黑色素瘤中的作用机制 转移该目的旨在鉴定负责IGF2BP 1功能的IGF2BP 1靶点， 黑素瘤转移为此，我们提出了几种互补的方法来执行无偏 IGF2BP 1参与黑色素瘤转移的分析。我们的方法将包括：i）分析或RNA 在体内黑色素瘤中IGF2BP1功能抑制后下调; ii）体内分析能够抑制IGF2BP1功能的cDNA， 克服由IGF2BP 1下调引起的黑素瘤转移的抑制; iii）分析 黑色素瘤中IGF2BP1抑制后半衰期和mRNA表达;和iv）mRNA的鉴定 在转移性黑素瘤细胞中与IGF2BP1直接相互作用。具体目标2。分析IGF2BP1的作用 EV介导的黑色素瘤进展。本研究旨在全面评价 IGF2BP1在黑色素瘤EV中的作用，特别是因为它与它们在肿瘤进展中的作用有关。我们的分析将包括 阐明IGF2BP1在EV介导的转移前小生境形成中的作用;其在受体EV摄取中的作用 体外和体内细胞; RNA转移到受体细胞的效率; EV数量、大小、组成分析 和货物;和验证我们的研究结果在EV分离的黑色素瘤患者的血清。影响： 成功完成这项研究，我们将：（一）了解IGF2BP 1的贡献和机制 （B）更好地理解IGF2BP 1及其靶基因如何参与黑色素瘤进展; 黑色素瘤转移;和（c）建立IGF2BP 1作为黑色素瘤患者的新治疗靶点。的 提出的研究，结合联合收割机细胞和分子生物学，生物信息学，技术开发， 尖端的体内实验将提供深入了解IGF2BP 1和其靶点之间的相互作用 分子以及这些分子如何促进黑色素瘤进展。最终，本研究中获得的数据 为黑色素瘤的新疗法铺平道路的潜力。

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.