Aoki M, Morishita R, Higaki J, Moriguchi A, Kida I, Hayashi S, Matsushita H, Kaneda Y, Ogihara T
Department of Geriatric Medicine, Osaka University Medical School, Japan.
Biochem Biophys Res Commun. 1997 Feb 24;231(3):540-5. doi: 10.1006/bbrc.1996.5762.
Although antisense strategy has been at the center of interest in gene therapy, little is known about application of this strategy to cardiac diseases because of the lack of a suitable delivery method into the heart. Therefore, we compared the transfection efficiency of antisense oligodeoxynucleotides (ODN) using HVJ-liposome method and direct transfer in in vivo transfer into heart. To investigate the cellular fate and localization of ODN, transfer of "naked" FITC-labeled antisense ODN or ODN enwrapped in HVJ-liposome complex were examined. Following in vivo transfer using direct injection as well as in vitro transfer, fluorescence rapidly disappeared within 1 day, whereas transfer by HVJ-liposome method resulted in sustained fluorescence localized in the nucleus for at least 1 week. Measurement of fluorescence also demonstrated a significantly higher level in myocardium transfected by HVJ-liposome method than direct transfer. The present study demonstrated that HVJ-liposome method is more efficacious for ODN delivery by prolongation of half-life of ODN, suggesting its usefulness for gene therapy in cardiac diseases.
尽管反义策略一直是基因治疗关注的核心，但由于缺乏将其导入心脏的合适递送方法，对于该策略在心脏病治疗中的应用了解甚少。因此，我们比较了使用HVJ-脂质体法和直接注射法将反义寡脱氧核苷酸（ODN）体内导入心脏的转染效率。为了研究ODN的细胞命运和定位，我们检测了“裸露的”异硫氰酸荧光素（FITC）标记的反义ODN或包裹在HVJ-脂质体复合物中的ODN的转移情况。通过直接注射进行体内转移以及体外转移后，荧光在1天内迅速消失，而通过HVJ-脂质体法转移则导致荧光持续存在于细胞核中至少1周。荧光测量还显示，通过HVJ-脂质体法转染的心肌中的荧光水平明显高于直接注射法。本研究表明，HVJ-脂质体法通过延长ODN的半衰期更有效地递送ODN，提示其在心脏病基因治疗中的应用价值。
