Sleeping Beauty-Mediated microRNA Therapeutics for Metastatic Colorectal Cancer

睡美人介导的 microRNA 治疗转移性结直肠癌

基本信息

批准号：
8689231
负责人：
R. Scott McIvor
金额：
$ 16.53万
依托单位：
UNIVERSITY OF MINNESOTA
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-04-01 至 2016-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8689231
关键词：
3&apos Untranslated Regions Address Affect Angiogenesis Inhibitors Animal Cancer Model Animal Model Binding Caenorhabditis elegans Cancer Etiology Cancer cell line Canis familiaris Catheters Cells Cessation of life Clinical Trials Colorectal Cancer Colorectal Neoplasms Cultured Cells Cyclophosphamide DNA delivery Data Development Diagnosis Diffusion Disease Effectiveness Europe Excision Firefly Luciferases Fluorouracil Functional RNA Gap Junctions Gene Delivery Gene Expression Gene Targeting Gene Transfer Generations Growth HCT116 Cells Hepatic Hepatocyte High Density Lipoproteins Human Indium Individual Length Link Liver Luciferases Malignant Neoplasms Mediating Mediator of activation protein Messenger RNA Metastatic Neoplasm to the Liver Metastatic to MicroRNAs Molecular Mus Neoplasm Metastasis Normal Cell Normal tissue morphology Nucleotides Nude Mice Oncogenic Operative Surgical Procedures Pathway interactions Patients Pattern Penetration Play Positioning Attribute Proteins Reporter Reporter Genes Research Project Grants Role Site Sleeping Beauty Stromal Neoplasm System Techniques Testing Therapeutic Time Tissues Translations Treatment Protocols Tumor Suppression Tumor Suppressor Proteins United States Xenograft Model Antitumor Assays advanced disease base bioluminescence imaging cancer therapy cancer type chemotherapy gain of function in vivo in vivo Model intercellular communication metastatic colorectal mouse model neoplastic cell novel therapeutic intervention outcome forecast overexpression pre-clinical public health relevance success therapeutic gene tumor tumor xenograft tumorigenesis vector

项目摘要

DESCRIPTION (provided by applicant): Colorectal cancer is one of the leading causes of cancer death worldwide, with the liver being the most common and critical site for development of colorectal cancer metastases. Surgical resection remains the only curative hope for patients with disease that is metastatic to the liver. However, 85% of these patients are ineligible for resection, with the 5-year survival for inoperative patients ranging from 0-5%. MicroRNAs (miRNAs) are non-coding RNAs approximately 20 nucleotides in length that exert their regulatory effects by binding to imperfect complementary sites predominantly located within the 3'UTR of their mRNA targets. Their causal role in promoting tumorigenesis, invasion and metastases has only recently come to light. Almost all cancer types show aberrant expression of miRNAs, with patterns of overexpression or, more commonly, underexpression in tumor cells. We hypothesize that these miRNAs constitute valid targets for the development of molecular therapies against metastatic disease. Here we propose to use the Sleeping Beauty transposon system to achieve extended and high level liver-directed expression of both tumor suppressor miRNAs as well as antimiRs against oncogenic miRNAs in the treatment of human colorectal cancer metastatic to the liver. Although it is well established that miRNAs can be transferred horizontally by a number of different mechanisms, in vivo data demonstrating exogenous miRNA transfer from normal cells to tumor cells in an animal model of cancer are lacking. In Aim 1, we have devised an approach to quantitatively evaluate intercellular transfer of miRNA from normal liver tissue into human tumor xenografts, assaying for miRNA delivery to an mRNA target by the effect on luciferase reporter gene expression in tumors. In Aim 2 we will test the effectiveness of tumor suppressive miRNAs (miR-34a, miR-9) as well as targeting an oncogenic miRNA (miR-21) both singly and in combination against human colorectal tumor xenografts in nude mice. Results from these studies will provide fundamental experimental support for therapeutic microRNA transfer from normal tissues into tumor cells in an in vivo model of metastatic disease. They will also provide preclinical support for an miRNA-based antitumor strategy against metastatic colorectal cancer that is potentially applicable to the treatment of an cancer that is metastatic to the liver.

描述（由申请人提供）：结直肠癌是全世界癌症死亡的主要原因之一，肝脏是结直肠癌转移最常见和最关键的部位。手术切除仍然是肝脏转移性疾病患者唯一的治愈希望。然而，其中 85% 的患者不适合切除，不手术患者的 5 年生存率为 0-5%。 MicroRNA (miRNA) 是长度约为 20 个核苷酸的非编码 RNA，通过结合主要位于其 mRNA 靶标 3'UTR 内的不完美互补位点来发挥调节作用。它们在促进肿瘤发生、侵袭和转移中的因果作用直到最近才被发现。几乎所有癌症类型都表现出 miRNA 的异常表达，在肿瘤细胞中存在过度表达或更常见的表达不足的模式。我们假设这些 miRNA 构成了开发针对转移性疾病的分子疗法的有效靶点。在这里，我们建议使用睡美人转座子系统来实现肿瘤抑制 miRNA 以及针对致癌 miRNA 的 antimiR 的延长和高水平肝脏定向表达，以治疗转移至肝脏的人类结直肠癌。尽管已经明确 miRNA 可以通过多种不同的机制水平转移，但缺乏证明外源 miRNA 在癌症动物模型中从正常细胞转移到肿瘤细胞的体内数据。在目标 1 中，我们设计了一种定量评估 miRNA 从正常肝组织到人类肿瘤异种移植物的细胞间转移的方法，通过对肿瘤中荧光素酶报告基因表达的影响来分析 miRNA 向 mRNA 靶标的传递。在目标 2 中，我们将测试肿瘤抑制 miRNA（miR-34a、miR-9）以及靶向致癌 miRNA（miR-21）的有效性，无论是单独还是联合使用，在裸鼠中对抗人类结直肠肿瘤异种移植物。这些研究的结果将为转移性疾病体内模型中的治疗性 microRNA 从正常组织转移到肿瘤细胞中提供基础实验支持。他们还将为基于 miRNA 的转移性结直肠癌抗肿瘤策略提供临床前支持，该策略可能适用于治疗肝转移性癌症。