Role and mechanisms of IGF2BP1 in melanoma pathogenesis

黑色素瘤是最致命的癌症之一。管理不善的主要原因 黑色素瘤是我们对黑色素瘤生物学发病机制的认识不足。最近，对mRNA的调控 细胞质中的命运是控制基因表达的主要机制。RNA结合蛋白可提供 参与肿瘤进展的通路调控的另一个维度。IGF2BP1是一种RNA结合蛋白 调节其信使核糖核酸靶标的稳定性、定位和翻译的蛋白质。我们之前已经表明， RNA结合蛋白IGF2BP1在黑色素瘤中高表达，但IGF2BP1在发病机制中的作用 黑色素瘤的发病机制尚未阐明。我们在不同小鼠模型上的新的初步数据表明IGF2BP1 影响黑色素瘤的转移，其在人类黑色素瘤中的高表达与不良的临床预后相关。 根据我们之前的研究和新的令人兴奋的观察结果，我们假设IGF2BP1在 黑色素瘤的进展和转移。为了验证这一假设，我们提出了以下独立但高度独立的假设 相互关联的特定靶点：特定靶点1.分析IGF2BP1在黑色素瘤中的作用机制 转移。该目标旨在确定负责IGF2BP1功能的IGF2BP1靶 黑色素瘤转移。为此，我们提出了几种互补的方法来执行不偏不倚的 IGF2BP1参与黑色素瘤转移的研究我们的方法将包括：i)分析或RNA 体内抑制黑色素瘤IGF2BP1功能下调；ii)体内分析能够 克服IGF2BP1下调导致的黑色素瘤转移抑制；iii)变化分析 IGF2BP1抑制黑色素瘤的半衰期和mRNA的表达；以及iv)mRNAs的鉴定 在转移性黑素瘤细胞中直接与IGF2BP1相互作用。具体目的2.分析IGF2BP1的作用 在EV介导的黑色素瘤进展中。这一目的是为了全面评估 IGF2BP1在黑色素瘤EV中的作用，特别是当它与它们在肿瘤进展中的作用有关时。我们的分析将包括 IGF2BP1在EV介导的转移前生态位形成中的阐明及其在EV受体摄取EV中的作用 体外和体内细胞；核糖核酸转移到受体细胞的效率；EV数量、大小、成分的分析 以及对我们从黑色素瘤患者血清中分离的EV的研究结果的验证。影响：打开 成功完成这项研究后，我们将：(A)了解IGF2BP1的贡献和机制 参与黑色素瘤的进展；(B)更好地了解IGF2BP1及其靶基因如何参与 (C)将IGF2BP1确立为黑色素瘤患者的新治疗靶点。这个 建议的研究结合了细胞和分子生物学、生物信息学、技术开发和 尖端的体内实验将为IGF2BP1与其靶点之间的相互作用提供深入的见解 以及这些分子如何促进黑色素瘤的进展。最终，这项研究中获得的数据 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.