DEVELOPMENTAL AND HORMONAL REGULATION OF APOLIPOPROTEIN B GENE EXPRESSION

载脂蛋白 B 基因表达的发育和激素调节

• 批准号: 3748219
• 负责人: A P PATTERSON
• 金额: --
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/3748219
• 关键词: HeLa cells; RNA binding protein; apolipoproteins; atherosclerosis; atherosclerotic plaque; cytokine receptors; enzyme deficiency; gene expression; genetic promoter element; genetically modified animals; hormone regulation /control mechanism; human genetic material tag; human subject; interleukin 4; laboratory mouse; laboratory rat; lipoprotein lipase; macrophage; messenger RNA; molecular cloning; posttranscriptional RNA processing; protein sequence; receptor expression; tissue /cell culture; triiodothyronine

We characterized apolipoprotein B (apoB) expression during rat development, and reported the first in vivo correlation between apoB mRNA editing activity and the expression of an RNA editing protein (REPR). We demonstrated that the normal development of apoB mRNA editing requires thyroid hormone (T3), and that the T3 modulates REPR mRNA levels. We cloned and characterized the tissue specific expression of a human homologue to rat REPR; this putative human form of REPR gene contains a zinc-finger domain with 100% homology to the transcription factor YY-1. Current studies include: (1) defining the regulatory elements of the rat REPR gene promoter; (2) sequencing the entirety of the human REPR homologue; (3) examining the in vitro expression of REPR in liver , intestinal, and HeLa cell lines. In collaboration with the Molecular Disease Branch of NHLBI, we are developing transgenic and knockout REPR mouse models to examine the physiologic consequences of REPR over and under expression. In our clinical studies (1) we examined intestinal apoB mRNA editing in a cohort of 8 patients with premature coronary artery disease and 15 normal volunteers, and reported the first example of a defective mRNA intestinal editing associated with premature artherosclerosis; (2) we isolated mRNA from the adipose tissue of a child with lipoprotein lipase deficiency (LPL) and several normal patients, and collaborated in the quantitation of LPL transcription; we have observed that this case of LPL deficiency is secondary to a post-transcriptional defect in LPL expression. Finally we have initiated a collaboration with another section of LMTB in the characterization of IL-4 receptor expression in normal vs. activated macrophages (foam cells) in the artherosclerotic plaque.

本研究观察了大鼠肝移植过程中载脂蛋白B（apoB）的表达， 发展，并报道了载脂蛋白B mRNA之间的第一个体内相关性 编辑活性和RNA编辑蛋白（REPR）的表达。 我们证明apoB mRNA编辑的正常发展需要 甲状腺激素（T3），T3调节REPR mRNA水平。 我们 克隆并表征了人的组织特异性表达 与大鼠REPR同源;这种假定的人类形式的REPR基因含有 与转录因子YY-1具有100%同源性的锌指结构域。 目前的研究包括：（1）确定大鼠的调控元件 REPR基因启动子;（2）对人REPR基因的全部序列进行测序， 同源物;（3）检测REPR在肝脏中的体外表达， 肠和HeLa细胞系。 与Molecular合作 NHLBI疾病分支，我们正在开发转基因和敲除REPR 小鼠模型来检查REPR的生理后果， 在表达下。 在我们的临床研究中（1），我们检查了肠道 apoB mRNA编辑在8例早发冠状动脉粥样硬化性心脏病患者中的应用 动脉疾病和15名正常志愿者，并报告了第一例 与早产相关的缺陷mRNA肠编辑 （2）我们从一名儿童的脂肪组织中分离出mRNA， 脂蛋白脂酶缺乏症（LPL）患者和几名正常患者， 合作定量LPL转录;我们观察到 这种LPL缺陷是继发于转录后 LPL表达缺陷。 最后，我们开始与 LMTB在IL-4受体表征中的另一部分 正常与活化巨噬细胞（泡沫细胞）中的表达 动脉粥样硬化斑块