Secreted RNA during CRC progression biogenesis function and clinical markers

CRC进展过程中分泌的RNA生物发生功能和临床标志物

• 批准号: 8927107
• 负责人: Robert J. Coffey
• 金额: $ 7.85万
• 依托单位: VANDERBILT UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-09-01 至 2018-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8927107
• 关键词: Address; Affect; Behavior; Biogenesis; Biological; Biological Assay; Biological Markers; Biological Models; Biology; Blood; Blood Circulation; Cardiovascular system; Cell physiology; Cells; Clinical Markers; Colon Carcinoma; Colorectal Cancer; Conditioned Culture Media; Data; Development; Disease; Disease Progression; Distant; Endocytosis; Environment; Extracellular Fluid; Flow Cytometry; Fluorescence; Genes; Goals; Health; Human; Hybrids; Immune; Individual; KRAS2 gene; Large Intestine Carcinoma; Lead; Liquid substance; Malignant Epithelial Cell; Malignant Neoplasms; Messenger RNA; MicroRNAs; Molecular; Mus; Mutation; Oncogenic; Pathway interactions; Physiology; Plasma; Population; Process; Proteins; RNA; RNA Processing; RNA Sequences; Research; Role; Sequence Analysis; Signal Pathway; Signal Transduction; Sorting - Cell Movement; Specificity; Stem cells; System; Technology; Testing; Tissues; Travel; Vesicle; Whole Organism; base; cancer cell; deep sequencing; extracellular; in vivo Model; insight; interest; late endosome; mutant; neoplastic cell; new technology; new therapeutic target; protein transport; research study; response; scaffold; tool; trafficking; tumor; tumor progression; tumor xenograft; uptake

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.

描述（由申请人提供）：该提议的一个主要前提是，生物流体中存在的RNA代表了进入多种组织中存在的各种细胞过程的窗口， 释放这些RNA。如果我们能够理解这些RNA是如何在细胞外结束的，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的反应，比如在免疫信号传导过程中或在发育或干细胞过程中存在的信号等）。然后我们就可以对这些可获得的物质进行分析，从而提供整个生物体的生理信息。我们已经发现，发生在结直肠癌中的致癌KRAS突变可以调节外泌体中肿瘤细胞分泌的miRNA、mRNA和长RNA。在这些建议中，我们将确定肿瘤细胞分泌的细胞外泌体RNA（eRNAs-project 1）和那些进入循环系统的包装RNA（cRNAs-project 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RNA测序分析来完成。基于这些信息，我们将找到与这些RNA相关的序列，这些RNA将它们运输到外泌体，并测试特定RNA加工和运输蛋白在将这些RNA递送到外泌体中的作用，从而导致它们分泌出细胞（项目2）。我们将挑选10个RNA-外泌体靶点，将其贩运到体内模拟的血液系统中;测试RNA靶向序列和RNA相关蛋白在将这些RNA递送到血液中的作用（项目1）。最后，将在原发性人类肿瘤异种移植物中测试这些RNA进入循环系统的运输以及致癌性结直肠癌突变如何影响这些RNA及其相关的运输机制（项目1）。这些实验对生物标志物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RNA分泌的其他生物学范式中的功能创造了工具。我们相信，这些特定的RNA运输机制将在体内其他组织中发挥作用，并与多个领域具有普遍相关性。