Regulated expression of a foreign gene targeted to the ischaemic myocardium.
Regulated expression of a foreign gene targeted to the ischaemic myocardium.
复制标题
针对缺血心肌的外源基因的调节表达。
DOI：
10.1016/s0008-6363(97)00158-2
复制
发表时间：
1997
影响因子：
10.8
通讯作者：
Webster,KA
中科院分区：
文献类型：
--
作者：
Prentice,H;Bishopric,NH;Hicks,MN;Discher,DJ;Wu,X;Wylie,AA;Webster,KA
目的：转移外源基因的调节表达可能是基因治疗的一个重要特征。因为冠状动脉疾病通常涉及间歇性心肌缺血，然后是正常心脏功能的时期，所以可能有必要直接响应缺血相关信号来调节推定的治疗/心脏保护基因的表达。当前研究的目的是开发基因调控组件的组合，可用于将产物靶向心肌并将基因的表达限制在缺血活动期间。方法：构建表达质粒，其中含有肌肉特异性启动子和与报告基因连接的缺氧反应增强子元件。在培养细胞中瞬时表达或将 DNA 直接注射到兔心肌后，测量这些构建体对缺氧或实验性缺血的调节。结果：紧邻最小肌肉特异性 α-肌球蛋白重链启动子上游的一组缺氧反应元件通过体外缺氧和体内缺血赋予该启动子有效的正向调节。缺血诱导持续至少4小时，并在8小时内恢复到基线水平。结论：缺氧响应调节元件与掺入适当载体系统的弱组织限制性启动子相结合，可以允许治疗基因在缺血心肌中受控表达。
Objectives:Regulated expression of transferred foreign genes may be an important feature of gene therapy. Because coronary artery disease often involves intermittent myocardial ischaemia followed by periods of normal cardiac function it will probably be necessary to regulate the expression of putative therapeutic/cardioprotective genes directly in response to ischaemia-associated signals. The objectives of the current study were to develop a combination of gene regulatory components that can be used to target a product to the myocardium and limit the expression of the gene to periods of ischaemic activity.Methods:Expression plasmids were constructed containing muscle-specific promoters and hypoxia-responsive enhancer elements linked to a reporter gene. The regulation of these constructs by hypoxia or experimental ischaemia was measured following transient expression in cultured cells or after direct injection of DNA into the rabbit myocardium.Results:A single set of hypoxia response elements placed immediately upstream of the minimal muscle-specific α-myosin heavy chain promoter conferred potent positive regulation of this promoter by hypoxia in vitro and by ischaemia in vivo. Induction by ischaemia persisted for at least 4 h and returned to the baseline level within 8 h.Conclusions:Hypoxia responsive regulatory elements, in combination with weak tissue-restricted promoters incorporated into an appropriate vector system may allow controlled expression of a therapeutic gene in ischaemic myocardium.