Transgenic rat overexpressing miR-21 in vascular smooth muscle cells:functional i

转基因大鼠在血管平滑肌细胞中过度表达 miR-21：功能性 i

• 批准号: 7789816
• 负责人: Junming Yue
• 金额: $ 23.24万
• 依托单位: UNIVERSITY OF TENNESSEE HEALTH SCI CTR
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-01-12 至 2011-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7789816
• 关键词: 1-Phosphatidylinositol 3-Kinase; 3' Untranslated Regions; Address; Age; Animal Model; Apoptosis; BCL2 gene; Biological; Biological Process; Blood Vessels; Body Size; Cardiac; Cardiovascular Diseases; Carotid Arteries; Carotid Artery Injuries; Defect; Development; Diagnosis; Disease; Exhibits; Functional RNA; Gene Expression; Genes; Grant; Growth; Heart Hypertrophy; Hereditary Disease; Human; Hyperplasia; Hypertrophy; Injury; Knockout Mice; Lentivirus Vector; Limb structure; Malignant Neoplasms; Messenger RNA; Methods; MicroRNAs; Microscopic; Mitogen-Activated Protein Kinases; Modeling; Molecular; Mus; Oncogenic; PTEN gene; Paralysed; Pathway interactions; Phenotype; Physiological; Play; Proteins; Proto-Oncogene Proteins c-akt; Pulmonary Hypertension; Rattus; Regulation; Reporting; Role; Signal Pathway; Signal Transduction; Small RNA; Smooth Muscle; Smooth Muscle Myocytes; Testing; Transgenic Organisms; Translations; Vascular Diseases; Virus Diseases; base; cardiovascular disorder therapy; injured; insight; molecular phenotype; mouse model; neointima formation; overexpression; promoter; public health relevance; transgene expression

DESCRIPTION (provided by applicant): MicroRNAs (miRNAs) are a new class of non-coding small RNAs that negatively regulate gene expression by either degrading mRNA or inhibiting protein translation. miRNAs have been shown to play very important roles in a variety of diseases, such as cancers, viral infection, genetic disorders, and cardiovascular diseases. Recently we found that miR-21, an oncogenic miRNA gene was aberrantly overexpressed in a variety of cancers, was also upregulated in the mouse cardiac hypertrophy model and rat carotid artery balloon injury model. Knockdown of miR-21 induced apoptosis in vascular smooth muscle cells and in the rat carotid artery balloon-injured model by targeting the 3'untranslated region (UTR) of the phosphatase and tensin homolog (PTEN) gene and indirectly regulated Bcl-2 gene expression in vascular smooth muscle cells (VSMCs). To further understand the molecular mechanism by which miR-21 regulates gene expression on the important signaling pathways, which play pivotal roles in vascular diseases, we constructed a lentiviral vector in which the miR-21 gene was driven by the rat smooth muscle cell specific promoter (rSM22) and generated a transgenic rat model in which miR-21 is overexpressed in VSMCs. We found that several transgenic rat founders display consistent phenotypes: growth retardation and slightly paralyzed hind limbs. This transgenic rat model will certainly provide insight to understanding miRNA functions in cardiovascular diseases and will help us further evaluate potential applications of miRNA in diagnosing and test miRNA based therapy for cardiovascular diseases. PUBLIC HEALTH RELEVANCE: This proposal is to characterize transgenic rats expressing miR-21 using lentiviral vector to address the biological functions of miRNAs by targeting miR-21 gene into vascular smooth muscle cells. The phenotype and molecular mechanisms underlying the phenotypes will also be examined using this transgenic rat models by investigating how miR-21 is involved in the posttranscriptional regulation.

描述(申请人提供)：microRNAs(MiRNAs)是一类新的非编码小RNA，通过降解mRNA或抑制蛋白质翻译来负向调节基因的表达。MiRNAs已被证明在多种疾病中发挥着非常重要的作用，如癌症、病毒感染、遗传疾病和心血管疾病。最近我们发现miR-21是一种致癌的miRNA基因，在多种癌症中异常过表达，在小鼠心肌肥厚模型和大鼠颈动脉球囊损伤模型中也上调。在大鼠颈动脉球囊损伤模型中，miR-21通过靶向PTEN基因的3‘非翻译区(UTR)，间接调节血管平滑肌细胞(VSMCs)中Bcl2基因的表达，从而诱导血管平滑肌细胞和大鼠颈动脉球囊损伤模型中的细胞凋亡。为了进一步了解miR-21调控在血管疾病中起关键作用的重要信号通路上基因表达的分子机制，我们构建了由大鼠血管平滑肌细胞特异性启动子(RSM22)驱动miR-21基因的慢病毒载体，并建立了在VSMCs中高表达miR-21的转基因大鼠模型。我们发现，几个转基因大鼠创始人表现出一致的表型：生长迟缓和后肢轻微瘫痪。这一转基因大鼠模型将为了解miRNA在心血管疾病中的功能提供一定的见解，并将有助于我们进一步评估miRNA在心血管疾病诊断和测试基于miRNA的治疗中的潜在应用。 公共卫生相关性：这项建议是利用慢病毒载体鉴定表达miR-21的转基因大鼠，通过将miR-21基因靶向血管平滑肌细胞来研究miRNAs的生物学功能。通过研究miR-21如何参与转录后调控，也将使用这种转基因大鼠模型来研究表型和潜在的分子机制。