Selective tropism of the recombinant adeno-associated virus 9 serotype for rat cardiac tissue
Selective tropism of the recombinant adeno-associated virus 9 serotype for rat cardiac tissue
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DOI：
10.1002/jgm.1404
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发表时间：
2010-01-01
影响因子：
3.5
通讯作者：
Katovich, Michael J.
中科院分区：
文献类型：
--
作者：
Qi, Yanfei;Liu, Xuan;Katovich, Michael J.
背景心脏基因转移可能成为治疗心脏病的一种新方法。重组腺相关病毒（recombinant adeno-associated virus，rAAV）血清型众多，但对心肌组织的嗜性不同。方法通过体外和体内实验比较rAAV-2、5、7、8和9的心肌嗜性。对于体外研究，使用rAAV-2、5、7、8或9的10（7）个载体基因组（vg）来扩增大鼠新生心肌细胞（RNCM）和成纤维细胞（RNCF）。对于体内研究，通过相对非侵入性的心内注射在5日龄大鼠中施用4 X 10（10）vg的rAAV-2、5、7、8或9和4 X 10（11）vg的rAAV 8或9。结果rAAV 9和rAAV 2在RNCM中的转导效率在病毒转导后第3天最高。只有rAAV 2在RNCF中引起任何转导。体内实验结果表明，rAAV 9> rAAV 8> rAAV 7> rAAV 2 = rAAV 5。肝脏中的转导效率顺序为：rAAV 2> rAAV 5> rAAV 7> rAAV 8> rAAV 9。注射更高剂量（4 × 10（11）vg）的rAAV 9在心脏中提供比rAAV 8更广泛和高度心脏选择性的GFP表达。零到最小的表达绿色荧光蛋白被发现在肺和肾脏的两个剂量的所有rAAV血清型utilized.Conclusions，在本研究中获得的结果表明，rAAV 9提供了最具选择性和稳定的转导效率在心脏组织中，这种表达主要表现在心肌细胞。版权所有（C）2009约翰威利父子有限公司
Background Cardiac gene transfer may serve as a novel therapeutic approach for heart disease. Numerous serotypes of recombinant adeno-associated virus (rAAV) have been identified with variable tropisms to cardiac tissue.Methods Both in vitro and in vivo experiments were undertaken to compare cardiac tropisms of rAAV-2, 5, 7, 8 and 9. For the in vitro studies, 10(7) vector genome (vg) of rAAV-2, 5, 7, 8 or 9 were used to transduce both rat neonatal cardiac myocytes (RNCM) and fibroblasts (RNCF). For the in vivo studies, 4 x 10(10) vg of rAAV-2, 5, 7, 8 or 9, and 4 x 10(11) vg of rAAV8 or 9 were administered in 5-day-old rats via a relatively non-invasive intracardiac injection. One and two months post-administration, green fluorescent protein (GFP) expression in tissues was visualized and GFP mRNA was quantified by the real-time polymerase chain reaction.Results At 3 days post-viral transduction, rAAV9 and rAAV2 produced the highest transducing efficiency in RNCM. Only rAAV2 elicited any transduction in the RNCF. The results obtained in vivo indicated that the order for transduction efficiency in the heart was: rAAV9 > rAAV8 > rAAV7 > rAAV2 = rAAV5. The transduction efficiency order in the liver was: rAAV2 > rAAV5 > rAAV7 > rAAV8 > rAAV9. Injection of a higher dose (4 x 10(11) vg) of rAAV9 provided more widespread and highly cardiac-selective GFP expression in the heart than rAAV8. Zero to minimal expression of GFP was found in the lung and kidney for both doses of all rAAV serotypes utilized.Conclusions Collectively, the results obtained in the present study suggest that rAAV9 provides the most selective and stable transduction efficiency in cardiac tissue, and this expression was primarily exhibited in cardiac myocytes. Copyright (C) 2009 John Wiley & Sons, Ltd.