The Role of Endothelial Derived miRNA17-92 in angiogenesis, an in vivo approach

内皮衍生的 miRNA17-92 在血管生成中的作用，一种体内方法

• 批准号: 8325242
• 负责人: Shira Landskroner-Eiger
• 金额: $ 5.39万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-12-01 至 2013-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8325242
• 关键词: 3' Untranslated Regions; Address; Amino Acids; Area; B-Cell Lymphomas; Back; Base Pairing; Biochemical; Biological Assay; Blood Vessels; Cardiovascular Diseases; Cardiovascular system; Cell Culture Techniques; Cell Proliferation; Cells; Cellular biology; Clinical Treatment; Data; Development; Disease; Employee Strikes; Endothelial Cells; Endothelium; Enzymes; Exhibits; Family; Gene Expression; Generations; Genome; Goals; Gold; Growth; Human; Immunoprecipitation; In Vitro; Individual; Ischemia; Label; Limb structure; Link; Luciferases; Lymphocyte Biology; Lysine; Malignant Neoplasms; Mass Spectrum Analysis; Measurement; Messenger RNA; MicroRNAs; Mind; Modeling; Morphogenesis; Mus; Oncogenic; Pathway interactions; Pattern; Phenotype; Physiological; Process; Proteomics; Regulator Genes; Research; Resistance; Retinal; Role; Seeds; System; Testing; Time; Tissues; Translations; Tube; Tumor Angiogenesis; Tumor Biology; Tumor-Derived; Untranslated RNA; Vascular Endothelial Growth Factors; Work; angiogenesis; base; bevacizumab; cell growth; cell type; frontier; human DICER1 protein; implantation; in vivo; in vivo Model; insight; migration; novel; novel therapeutic intervention; paracrine; protein expression; reconstitution; research study; response; restoration; stem; therapeutic target; tool; tumor

DESCRIPTION (provided by applicant): MicroRNAs (miRNAs) are short (;22nt) noncoding RNAs, that have emerged as important regulators of genes involved in the control of multiple physiological pathways, and often function to modulate or fine-tune cellular phenotypes such as development and differentiation. The role of miRNAs in specific processes involving vasculature patterning and defining miRNA targets in the vasculature system is an exciting, emerging new area with potential promise for novel therapeutic approaches in vascular related diseases. The miRNA17- 92 cluster (miR17-92) has been initially linked to oncogenic activity as demonstrated by its amplification in human B-cell lymphomas. A link between this miRNA cluster and angiogenesis was first indicated by a paracrine effect of tumor-derived miR17-92 on promoting angiogenesis. In this case, specific components of the cluster miR19 and miR18 were demonstrated to cause reduction of anti-angiogenic molecules suggesting a pro-angiogenic role for this cluster. In contrast, it has recently been proposed that this cluster has a cell intrinsic anti-angiogenic activity in endothelial cells in vitro. Importantly, we have shown that components of the miR17-92 can regulate positively aspects of VEGF induced cell growth and morphogenesis of endothelial cells (EC) in vitro. Herein, we propose to present the first in vivo platform to study the function of endothelial-derived miR17-92. The work proposed in this application will contribute to identifying the role of EC derived miRNA17-92 cluster in both physiological and pathophysiological angiogenesis in vivo. In addition we will characterize the mechanisms by which miR17-92 regulates angiogenesis by identifying novel targets for this cluster in the context of EC biology. We hypothesize that the miR17-92 cluster is a key regulator in vascular patterning and remodeling and that mice conditionally lacking miR17-92 in endothelial cells will exhibit abnormal angiogenic responses. Recent work by Tyler Jack's group has generated mice conditionally lacking the miR17-92 cluster. A major tool in this proposal is the generation of mice conditionally lacking the miR17-92 cluster in ECs. We have generated these mice and have exciting preliminary data indicating a potential developmental role for this cluster in regulating the timing and organization of the retinal vasculature. We will specifically examine the following: 1) The in vivo role of endothelial derived miR17-92 in models of angiogenesis 2) Identify and characterize the role of the EC miR17- 92 cluster as well as its individual "seed family" derivatives in in vitro aspects of angiogenesis. 3) Identify putative targets of the miR17-92 cluster in EC using SILAC analysis.

描述（由申请人提供）：microRNA（miRNA）是简短的（; 22nt）非编码RNA，它们已成为参与多个生理途径控制的基因的重要调节因子，并且通常在调节或微调细胞型（例如发育和差异化和差异化）中起作用。 miRNA在涉及脉管系统模式和脉管系统中定义miRNA靶标的特定过程中的作用是一个令人兴奋的，新兴的新领域，具有潜在的潜在的血管相关疾病治疗方法的希望。 miRNA17-92簇（miR17-92）最初与致癌活性有关，如它在人B细胞淋巴瘤中的扩增所证明的那样。该miRNA簇和血管生成之间的联系首先是由肿瘤衍生的miR17-92对促进血管生成的旁分泌作用表示的。在这种情况下，簇miR19和miR18的特定成分被证明会导致抗血管生成分子的降低，这表明该簇的促血管生成作用。相比之下，最近有人提出该簇在体外内皮细胞中具有细胞的固有性抗血管生成活性。重要的是，我们已经表明，miR17-92的成分可以在体外调节VEGF诱导的细胞生长和内皮细胞的形态发生（EC）的形态发生。本文中，我们建议介绍第一个研究内皮衍生的miR17-92功能的体内平台。本应用中提出的工作将有助于确定EC衍生的miRNA17-92簇在体内生理和病理生理血管生成中的作用。此外，我们将表征miR17-92通过在EC生物学背景下鉴定该簇的新靶标的MiR17-92调节血管生成的机制。我们假设MiR17-92簇是血管模式和重塑中的关键调节剂，并且在内皮细胞中有条件缺乏miR17-92的小鼠将表现出异常的血管生成反应。泰勒·杰克（Tyler Jack）小组的最新工作使小鼠有条件地缺乏Mir17-92簇。该提案中的一个主要工具是有条件地缺乏EC中的miR17-92簇的小鼠。我们已经产生了这些小鼠，并拥有令人兴奋的初步数据，这表明该集群在调节视网膜脉管系统的时机和组织中具有潜在的发展作用。我们将专门检查以下内容：1）内皮衍生的miR17-92在血管生成模型中的体内作用2）识别和表征EC miR17- 92簇的作用，以及其在血管生成的体外方面的单个“种子家族”衍生物。 3）使用SILAC分析确定EC中MIR17-92簇的假定靶标。