Rolling F, Nong Z, Pisvin S, Collen D
Center for Transgene Technology and Gene Therapy, Flanders Interuniversity Institute for Biotechnology, Leuven, Belgium.
Gene Ther. 1997 Aug;4(8):757-61. doi: 10.1038/sj.gt.3300465.
Gene transfer into the arterial wall may allow study of the role of specific genes in vascular pathophysiology and development of local gene therapies for vascular disorders. The feasibility of adeno-associated virus (AAV)-mediated gene transfer into isolated segments of normal and balloon-injured rat carotid arteries was studied using a recombinant AAV carrying CMVlacZ as a reporter gene. Approximately 10(6) and 10(7) infectious units (IU) of AAV were infused into 1 cm isolated segments of the carotid artery of 14 animals with the aid of a Silastic catheter and allowed to remain for 20 min. Animals were killed at different time-points after infection and arteries stained for beta-gal activity. Microscopic examination demonstrated comparable gene transfer into medial and adventitial cells, with significantly higher efficiency of transduction in injured as compared with normal vessels. High levels of in vivo beta-gal expression persisted for at least 30 days after gene transfer. Thus, AAV is capable of transducing media and adventitia of rat carotid arteries, suggesting that it may constitute a useful vector for arterial gene transfer and gene therapy protocols.
将基因导入动脉壁可能有助于研究特定基因在血管病理生理学中的作用，并开发针对血管疾病的局部基因疗法。使用携带CMV-lacZ作为报告基因的重组腺相关病毒（AAV），研究了AAV介导的基因转移到正常和球囊损伤的大鼠颈动脉分离段中的可行性。借助硅橡胶导管，将约10^6和10^7感染单位（IU）的AAV注入14只动物的1 cm颈动脉分离段中，并使其保留20分钟。在感染后的不同时间点处死动物，并对动脉进行β-半乳糖苷酶活性染色。显微镜检查表明，基因向中膜和外膜细胞的转移情况相当，与正常血管相比，损伤血管中的转导效率明显更高。基因转移后，体内β-半乳糖苷酶的高水平表达持续至少30天。因此，AAV能够转导大鼠颈动脉的中膜和外膜，这表明它可能构成动脉基因转移和基因治疗方案的有用载体。
