miR-126 Regulation of Tumor Progression

miR-126 对肿瘤进展的调节

基本信息

批准号：
8444708
负责人：
CALVIN J KUO
金额：
$ 36.63万
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/8444708
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 Functional RNA Future Gene Expression Generations Genes Genetic Grant Growth Factor 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 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 public health relevance response 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（由申请人提供）：microRNA（miRNA）是21-23个核苷酸长度的单链RNA分子，通过用靶mRNA的3'UTR退火，抑制翻译并诱导mRNA降解和脱甲基化来下调基因表达。对其mRNA靶标的miRNA识别不精确，可以在稳态期间或对刺激的响应期间对数百个靶标mRNA进行转录后调节。 MicroRNA miR-126代表表达分析时最丰富的内皮miRNA，并且在胚胎发生过程中以泛皮的方式表达。我们创建了缺乏miR-126的敲除小鼠，这些小鼠表现出与水肿，出血和血管生成延迟相关的50％胚胎致死性。在存活的miR-126 KO小鼠中，例如在角膜微量源分析中延迟成年血管生成。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的表达。先前尚未证明miRNA在肿瘤血管生成过程中的重要作用。当前的应用探讨了miR-126在肿瘤内皮中强的miR-126/eGFL7表达和乳腺癌MMTV-PYMT转基因模型中的初步数据中miR-126在肿瘤血管生成中的作用。 AIM 1研究了miR-126的组成型遗传缺失是否抑制肿瘤的进展和血管生成，并扩展了MMTV-PYMT模型中的存活率。 AIM 2在预先建立的MMTV-PYMT肿瘤中，通过临时有条件的miR-126 KO评估miR-126抑制的治疗潜力。有条件的miR-126 KO将通过两种Antagomirs以及新型的2'-F/甲氧酯抗MIR化学作为MMTV-PYMT模型中药物miR-126抑制的参考标准。此外，将比较通过遗传缺失或Antagomir/anti-MIR治疗的miR-126抑制作用，并与VEGF抑制作用进行比较。最后，AIM 3研究了通过内皮中室特异性缺失在乳腺肿瘤发生过程中miR-126作用的机制，在体外表征miR-126 KO内皮以及内皮尖端和茎细胞表型。总体而言，这些研究采用了严格特征的miR-126遗传基因敲除小鼠和新型Antagomir和2'-F/甲氧酯抗MIR治疗策略的互补方法，以探索内皮微生虫和肿瘤血管生成之间的第一个功能联系。在肿瘤血管生成和进展过程中，miR-126功能需求的证明将对未来的抗血管生成疗法的设计产生重大影响。