Secreted RNA during CRC progression biogenesis function and clinical markers

描述(申请人提供)：这项提议的一个主要前提是，存在于生物液中的RNA代表了了解存在于多种组织中的各种细胞过程的窗口，这些组织 释放这样的RNA。如果我们能够理解这些RNA是如何最终出现在细胞外的，哪些RNA在不同的条件下存在(比如这些RNA在某些疾病的存在下如何变化，或者对某些细胞信号做出反应，比如在免疫信号或者发育或干细胞过程中出现的信号等)。然后，我们可以对这些可获得的物质进行分析，提供有关整个有机体的生理信息。我们发现，发生在结直肠癌中的致癌KRAS突变可以调节肿瘤细胞分泌的miRNAs、mRNAs和长RNAs。在这些提案中，我们将确定肿瘤细胞分泌的胞外胞外RNA(eRNAs-项目1)和那些进入循环系统的包装RNA(CRNAs-项目2)的序列，以及这些RNA在癌症进展过程中如何变化。这将通过全面的RNAseq分析来完成。基于这些信息，我们将找到与这些RNA相关的序列，将它们运输到外体，并测试特定的RNA加工和运输蛋白在将这些RNA输送到外体，导致它们分泌到细胞外的作用(项目2)。我们将挑选10个RNA-外体靶标，它们将在体内模拟它们进入血液系统的运输；测试RNA靶向序列和RNA相关蛋白在将这些RNA输送到血液中的作用(项目1)。最后，这些RNA进入循环系统以及致癌的结直肠癌突变如何影响这些RNA及其相关的转运机制将在原发的人类肿瘤异种移植瘤中进行测试(项目1)。这些实验对生物标记物的研究具有深远的影响，为分析和理解这些分泌的RNA在癌症和其他RNA分泌的生物范例中的功能创造了工具。我们相信，这些特定的RNA转运机制将在体内其他组织中发挥作用，并与多个领域普遍相关。

DESCRIPTION (provided by applicant): A major premise of this proposal is that RNAs present in biofluids represent a window into various cellular processes that exist in multiple tissues that release such RNAs. If we can understand how these RNAs end up outside the cell, which RNAs are present under different conditions (like how these RNAs change in the presence of certain diseases or respond to certain cellular signals like those present during immune signaling or during developmental or stem cell processes etc.) then we can provide assays for these accessible substances that give information about the whole organism's physiology. We have found that oncogenic KRAS mutations that occur in colorectal cancer can regulate miRNAs, mRNAs and long RNAs secreted from tumor cells in exosomes. In these proposals we will determine the sequence of tumor cell secreted extracellular exosomal RNAs (eRNAs-project 1) and those packaged RNAs that travel into the circulatory system (cRNAs-project 2) and how these RNAs change during cancer progression. This will be done by comprehensive RNAseq analysis. Based on this information we will find sequences associated with these RNAs that traffic them to exosomes and test the role of specific RNA processing and trafficking proteins in delivering these RNAs to exosomes, leading to their secretion out of the cell (project 2). We will pick 10 RNA-exosomal targets that will have their trafficking into the blood system modeled in vivo; testing the roles of RNA targeting sequences and RNA-associated proteins in delivering these RNAs into the blood (project 1). Finally the trafficking of these RNAs into the circulatory system and how oncogenic colorectal cancer mutations affect these RNAs and their associated trafficking mechanisms will be tested in primary human tumor xenografts (project 1). These experiments have far reaching implications for biomarker research, creates tools for analyzing and understanding the function of these secreted RNAs in cancer and in other biological paradigms for RNA secretion. We are confident that these specific RNA trafficking mechanisms will be acting in other tissues in the body and are of general relevance to multiple fields.