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

• 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
• 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' 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

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(申请人提供)：结直肠癌是全球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疾病患者唯一的治愈希望。然而，85%的患者不符合手术切除的条件，非手术患者的5年生存率从0-5%不等。MicroRNAs(MiRNAs)是一种长度约为20个核苷酸的非编码RNA，通过与位于其mRNA靶标的3‘非编码区内的不完全互补位点结合而发挥其调节作用。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曝光。几乎所有的癌症类型都表现出miRNAs的异常表达，在肿瘤细胞中有过度表达或更常见的低表达模式。我们假设这些miRNAs是开发抗转移疾病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基因miRNAs和针对癌基因miRNAs的抗癌基因miRNAs在肝脏定向扩展和高水平表达，用于治疗转移到肝脏的人类结直肠癌。虽然众所周知，miRNAs可以通过许多不同的机制进行水平转移，但在癌症动物模型中，缺乏证明外源性miRNA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从正常肝组织到人肿瘤移植瘤的miRNA细胞间转移的方法，通过检测对肿瘤中荧光素酶报告基因表达的影响来检测miRNA向mRNA靶点的传递。在目标2中，我们将测试肿瘤抑制miRNAs(miR-34a，miR-9)以及靶向致癌miRNA(miR-21)对人结直肠癌裸鼠移植瘤的有效性。这些研究的结果将为体内转移疾病模型中从正常组织到肿瘤细胞的治疗性microRNA转移提供基本的实验支持。他们还将为基于miRNA的抗肿瘤策略提供临床前支持，以对抗转移性结直肠癌，该策略可能适用于肝转移癌的治疗。