Functional changes in secreted RNA biogenesis using in vivo colon tumor models

使用体内结肠肿瘤模型的分泌性 RNA 生物发生的功能变化

• 批准号: 9331322
• 负责人: Robert J. Coffey
• 金额: $ 43.49万
• 依托单位: VANDERBILT UNIVERSITY MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-09-01 至 2019-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9331322
• 关键词: Address; Affect; Biogenesis; Bioinformatics; Biological Assay; Biological Markers; Blood; Blood Circulation; Blood specimen; Carcinoma; Cell Line; Cell physiology; Cells; Clinical Markers; Codon Nucleotides; Colon Carcinoma; Colonic Neoplasms; Colorectal Cancer; Colorectal Neoplasms; Data; Disease; Distant; Environment; Event; Extracellular Fluid; Fluorescence; Gene Expression; Genetic; Genotype; Goals; Human; Image; In Vitro; Individual; Interest Group; KRAS2 gene; Label; Large Intestine Carcinoma; Mediating; Messenger RNA; MicroRNAs; Modeling; Mus; Mutation; Neoplasms; Normal Cell; Oncogenic; Organism; Paracrine Communication; Pathway interactions; Patients; Pattern; Physiological; Plasma; Pongidae; Primary Neoplasm; Production; Proteins; RNA; RNA Binding; Research; Resources; Route; Signal Transduction; Sorting - Cell Movement; System; Testing; Therapeutic; Time; Tissues; Travel; Tumor-Derived; Variant; Vesicle; Xenograft procedure; adenoma; base; behavior influence; cancer cell; exosome; experimental study; extracellular vesicles; in vivo; in vivo Model; late endosome; mouse model; mutant; mutational status; neoplastic cell; novel; novel strategies; protein profiling; protein transport; tool; trafficking; tumor; tumor progression; tumor xenograft

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. 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. This project (project 1) will determine what the mechanisms are by which these RNAs (cRNAs) enter the bloodstream using in vivo models and how these RNAs change with oncogenic mutations. This project will result in the creation of new tools to visualize and analyze such RNAs in real time and from blood samples in mouse models. We propose the following aims to address these questions. Aim 1. Determine circulating RNAs in human and mouse plasma that differ in normal individuals and those bearing colorectal tumors. We hypothesize that colorectal tumor cells secrete an altered composition of circulating RNAs (cRNAs) compared to normal cells. Further, many of these changes will be conserved from mouse to human CRC. The goal of this aim is to identify plasma cRNAs that are altered with tumor progression (advanced adenomas and carcinomas). Aim 2. Elucidate the pathway(s) by which tumor RNAs traffic into the circulation. Various mechanisms may contribute to CRC-induced cRNA alterations. In this aim, we will determine the route by which the exosome-associated miRNAs and long RNAs identified in Aim 1 enter the circulation. Aim 3. Determine the influence of tumor genotype on cRNAs using human primary tumor xenografts and determine the function of candidate trafficking proteins and interacting motifs in altering tumor RNA secretion. In conjunction with studies performed in Project 2, we will determine the mechanism by which RNAs are trafficked into exosomes and secreted into the blood. In this aim, we will determine how the genotype of human CRC xenografts affects exosome-associated cRNAs.

该提议的一个主要前提是，生物流体中存在的RNA代表了了解各种细胞的窗口。 这些过程存在于多种组织中，释放这些RNA。我们发现致癌KRAS 在结直肠癌中发生的突变可以调节肿瘤分泌的miRNAs、mRNAs和长RNA， 外泌体中的细胞。这个项目（项目1）将确定这些RNA的机制是什么， 使用体内模型研究cRNA进入血流的机制，以及这些RNA如何随着致癌突变而变化。 该项目将导致创建新的工具来可视化和分析这样的RNA在真实的时间和从 小鼠模型中的血液样本。我们提出以下目标来解决这些问题。 目标1.确定人类和小鼠血浆中的循环RNA，这些RNA在正常个体和结直肠肿瘤患者中存在差异。 我们假设，与正常细胞相比，结直肠肿瘤细胞分泌的循环RNA（cRNA）的组成发生了变化。此外，这些变化中的许多将从小鼠到人CRC是保守的。该目的的目标是鉴定随着肿瘤进展（晚期腺瘤和癌）而改变的血浆cRNA。 目标2.阐明肿瘤RNA进入循环的途径。 多种机制可能有助于CRC诱导的cRNA改变。为此，我们将确定Aim 1中鉴定的外泌体相关miRNA和长RNA进入循环的途径。 目标3.使用人原发性肿瘤异种移植物确定肿瘤基因型对cRNA的影响，并确定候选运输蛋白和相互作用基序在改变肿瘤RNA分泌中的功能。 结合项目2中进行的研究，我们将确定RNA被贩运到外泌体并分泌到血液中的机制。在这个目标中，我们将确定人类CRC异种移植物的基因型如何影响外泌体相关的cRNA。