Efficient and stable transduction of cardiomyocytes after intramyocardial injection or intracoronary perfusion with recombinant adeno-associated virus vectors
Efficient and stable transduction of cardiomyocytes after intramyocardial injection or intracoronary perfusion with recombinant adeno-associated virus vectors
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DOI：
10.1161/01.cir.99.2.201
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发表时间：
1999-01-19
期刊：
影响因子：
37.8
通讯作者：
Leiden, JM
中科院分区：
文献类型：
--
作者：
Svensson, EC;Marshall, DJ;Leiden, JM
背景-将重组基因导入心肌细胞有望治疗多种心血管疾病。先前使用直接注射质粒DNA或复制缺陷型腺病毒载体的基因转移方法分别受到低转导频率和由于免疫应答引起的瞬时转基因表达的限制。在这份报告中，我们测试了使用心肌内注射或冠状动脉内输注重组腺相关病毒（rAV）载体来编程小鼠体内心肌细胞中转基因表达的可行性。方法和结果-我们构建了包含LacZ基因的rAV巨细胞病毒（CMV）启动子的转录控制下（AAC（CMV-LacZ））。然后，我们将1 × 10（8）感染单位（IU）的该病毒注射到成年CD-1小鼠的左心室心肌中。对照心脏注射Ad（CMV-LacZ）腺病毒载体。在AAV（CMV-LacZ）注射后2、4和8周收获的心脏在大量心肌细胞中表现出稳定的β-半乳糖苷酶（β-gal）表达，而没有心肌炎症或心肌细胞坏死的证据。相比之下，Ad（CMV-LacZ）注射的心脏显示短暂的β-gal表达，这是不可检测的注射后4周。还通过冠状动脉用1.5 × 10（9）IU的AAV（CMV-LacZ）灌注移植的C57 BL/6小鼠心脏，并在2、4和8周后测定β-gal表达。β-Gal的表达在
Background-The delivery of recombinant genes to cardiomyocytes holds promise for the treatment of a variety of cardiovascular diseases. Previous gene transfer approaches that used direct injection of plasmid DNA or replication-defective adenovirus vectors have been limited by low transduction frequencies and transient transgene expression due to immune responses, respectively. In this report, we have tested the feasibility of using intramyocardial injection or intracoronary infusions of recombinant adeno-associated virus (rAAV) vectors to program transgene expression in murine cardiomyocytes in vivo.Methods and Results-We constructed an rAAV containing the LacZ gene under the transcriptional control of the cytomegalovirus (CMV) promoter (AAV(CMV-LacZ)). We then injected 1X10(8) infectious units (IU) of this virus into the left ventricular myocardium of adult CD-1 mice. Control hearts were injected with the Ad(CMV-LacZ) adenovirus vector. Hearts harvested 2, 4, and 8 weeks after AAV(CMV-LacZ) injection demonstrated stable beta-galactosidase (beta-gal) expression in large numbers of cardiomyocytes without evidence of myocardial inflammation or myocyte necrosis. In contrast, the Ad(CMV-LacZ)-injected hearts displayed transient beta-gal expression, which was undetectable by 4 weeks after injection. Explanted C57BL/6 mouse hearts were also perfused via the coronary arteries with 1.5X10(9) IU of AAV(CMV-LacZ) and assayed 2, 4, and 8 weeks later for beta-gal expression. beta-Gal expression was detected in