miR-126 Regulation of Tumor Progression

miR-126 对肿瘤进展的调节

基本信息

批准号：
8815103
负责人：
CALVIN J KUO
金额：
$ 39.11万
依托单位：
STANFORD UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2011
资助国家：
美国
起止时间：
2011-03-04 至 2016-02-28
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8815103
关键词：
3&apos Untranslated Regions Active Sites Adult Alleles Animal Model Benchmarking Biological Assay Blood Vessels Breast Cancer Model Cells Chemistry Clinical Practice Patterns Cornea Data Development Early Diagnosis Edema Embryo Embryonic Development Endothelial Cells Endothelium Evaluation Exhibits Future Gene Expression Generations Genes Genetic Grant Growth Factor Health Hemorrhage Homeostasis Human In Vitro Introns Investigation Knockout Mice Length Malignant Neoplasms Mammary Neoplasms Mammary Tumorigenesis Mass Spectrum Analysis Messenger RNA Methods MicroRNAs Modality Modeling Molecular Profiling Mouse Mammary Tumor Virus Mus Neoplasm Metastasis Neoplasms in Vascular Tissue Nucleotides PDGFB gene Phenotype Post-Transcriptional Regulation Prevention Preventive Intervention Proteomics RNA Reference Standards Regulation Repression Role Screening for cancer Stimulus Therapeutic Transgenic Model Transgenic Organisms Translations Tumor Angiogenesis Tumor Biology Untranslated RNA Validation Vascular Endothelial Growth Factors Work angiogenesis base breast tumorigenesis cancer therapy combinatorial design genetic analysis mRNA Transcript Degradation malignant breast neoplasm novel overexpression pre-clinical response targeted treatment tumor tumor progression tumorigenesis

项目摘要

DESCRIPTION (provided by applicant): MicroRNAs (miRNA) are single-stranded RNA molecules of 21-23 nucleotides length which down-regulate gene expression by annealing with the 3' UTR of target mRNAs, repressing translation and inducing mRNA degradation and de-adenylation. The imprecise miRNA recognition of their mRNA targets allows post-transcriptional regulation over hundreds of target mRNA during homeostasis or in response to stimuli. The microRNA miR-126 represents the most abundant endothelial miRNA upon expression profiling, and is expressed in a pan-endothelial fashion during embryogenesis. We have created knockout mice lacking miR-126 which exhibit 50% embryonic lethality associated with edema, hemorrhage, and angiogenic delay. In surviving miR-126 ko mice, adult angiogenesis is delayed for instance in corneal micropocket assays. Interestingly, miR-126 is present in intron 7 of a host gene, Egfl7. The miR-126 ko phenotype actually recapitulates previously described Egfl7 ko phenotypes, and previously described Egfl7 ko mice are now understood to have inadvertently disrupted miR-126 expression. An essential role for a miRNA during tumor angiogenesis has not been previously demonstrated. The current application explores the role of miR-126 in tumor angiogenesis based upon strong miR-126/Egfl7 expression in tumor endothelium and preliminary data in the MMTV- PyMT transgenic model of breast cancer. Aim 1 investigates whether constitutive genetic deletion of miR-126 inhibits tumor progression and angiogenesis and extends survival in the MMTV-PyMT model. Aim 2 evaluates the therapeutic potential of miR-126 inhibition through temporally conditional miR-126 ko with our floxed mouse allele in pre-established MMTV-PyMT tumors. Conditional miR-126 ko will serve as a reference standard for pharmacologic miR-126 inhibition in the MMTV-PyMT model via both antagomirs, as well as a novel 2'-F/methoxyester anti-miR chemistry. Further, miR-126 inhibition by either genetic deletion or antagomir/anti-mir treatment will be compared and combined with VEGF inhibition. Finally, Aim 3 investigates mechanisms of miR-126 action during breast tumorigenesis through compartment-specific deletion in endothelium, in vitro characterization of miR-126 ko endothelium, and endothelial tip- and stalk cell phenotypes. Overall, these investigations utilize complementary approaches of rigorously characterized miR-126 genetic knockout mice and novel antagomir and 2'-F/methoxyester anti-miR therapeutic strategies to explore the first functional linkages between an endothelial microRNA and tumor angiogenesis. The demonstration of a functional requirement for miR-126 during tumor angiogenesis and progression would have significant implications for design of future anti-angiogenic therapies.

描述（申请人提供）：MicroRNA（miRNA）是长度为 21-23 个核苷酸的单链 RNA 分子，通过与目标 mRNA 的 3' UTR 退火、抑制翻译并诱导 mRNA 降解和去腺苷酸化来下调基因表达。 miRNA 对 mRNA 靶标的不精确识别允许在稳态或响应刺激期间对数百个靶标 mRNA 进行转录后调节。 microRNA miR-126 代表表达谱中最丰富的内皮 miRNA，并在胚胎发生过程中以泛内皮方式表达。我们创建了缺乏 miR-126 的基因敲除小鼠，其表现出与水肿、出血和血管生成延迟相关的 50% 胚胎致死率。在存活的 miR-126 ko 小鼠中，成年血管生成被延迟，例如在角膜微袋测定中。有趣的是，miR-126 存在于宿主基因 Egfl7 的内含子 7 中。 miR-126 ko 表型实际上概括了先前描述的 Egfl7 ko 表型，并且先前描述的 Egfl7 ko 小鼠现在被认为无意中破坏了 miR-126 表达。 miRNA 在肿瘤血管生成过程中的重要作用尚未得到证实。本申请基于肿瘤内皮中 miR-126/Egfl7 的强表达和乳腺癌 MMTV-PyMT 转基因模型的初步数据，探讨了 miR-126 在肿瘤血管生成中的作用。目标 1 研究 miR-126 的组成型遗传缺失是否会抑制 MMTV-PyMT 模型中的肿瘤进展和血管生成并延长生存期。目标 2 评估通过暂时条件 miR-126 ko 与我们的 floxed 小鼠等位基因在预先建立的 MMTV-PyMT 肿瘤中抑制 miR-126 的治疗潜力。有条件的 miR-126 ko 将作为 MMTV-PyMT 模型中通过两种 antagomir 以及新型 2'-F/甲氧基酯抗 miR 化学物质抑制 miR-126 药理学的参考标准。此外，将比较通过基因删除或 antagomir/anti-mir 治疗对 miR-126 的抑制，并将其与 VEGF 抑制相结合。最后，目标 3 通过内皮区室特异性缺失、miR-126 ko 内皮的体外表征以及内皮尖端细胞和茎细胞表型来研究 miR-126 在乳腺肿瘤发生过程中的作用机制。总体而言，这些研究利用严格表征的 miR-126 基因敲除小鼠和新型 antagomir 和 2'-F/甲氧基酯抗 miR 治疗策略的互补方法来探索内皮 microRNA 与肿瘤血管生成之间的第一个功能联系。证明肿瘤血管生成和进展过程中 miR-126 的功能需求将对未来抗血管生成疗法的设计产生重大影响。