Keller L H, Vander Heide R, Pebbles K A, L'Ecuyer T J
Children's Hospital of Michigan, Detroit, MI 48201-2196, USA.
Pediatr Cardiol. 2000 May-Jun;21(3):223-7. doi: 10.1007/s002460010044.
Gene transfer techniques are increasingly being used to study blood vessel biology and develop models for gene therapy. To date, there are no reports of pulmonary vascular gene transfer performed either without adjunctive agents or during angioplasty. We sought to demonstrate the feasibility of recombinant gene transfer to the pulmonary artery of juvenile pigs using naked plasmid DNA delivered via percutaneous angioplasty techniques. Plasmid DNA directing the expression of beta-galactosidase was used to transfect one pulmonary artery while the contralateral vessel served as an untreated control. One delivery technique used a standard angioplasty balloon coated with a DNA-heparin mixture. The second involved infusion of DNA between an angioplasty balloon and a surrounding, microporous balloon. Vessels were harvested 3 or 4 days following gene delivery. Protein expression was demonstrated by immunohistochemical staining in transfected but not control vessels in 9/9 pigs. Vascular wall expression was limited to endothelial cells. Pulmonary artery gene transfer using naked plasmid DNA delivered via percutaneous angioplasty techniques is feasible. Using naked plasmid DNA removes the potential for toxicity associated with adjunctive agents. The described techniques provide novel methods for studying pulmonary vascular biology in vivo and for developing future gene therapies.
基因转移技术越来越多地用于研究血管生物学并开发基因治疗模型。迄今为止，尚无关于在没有辅助剂的情况下或在血管成形术期间进行肺血管基因转移的报道。我们试图证明使用经皮血管成形术技术递送的裸质粒DNA将重组基因转移至幼年猪肺动脉的可行性。将指导β-半乳糖苷酶表达的质粒DNA用于转染一条肺动脉，而对侧血管作为未处理的对照。一种递送技术使用涂有DNA-肝素混合物的标准血管成形术球囊。第二种方法是在血管成形术球囊和周围的微孔球囊之间注入DNA。在基因递送后3或4天收获血管。通过免疫组织化学染色在9/9只猪的转染血管而非对照血管中证实了蛋白质表达。血管壁表达仅限于内皮细胞。使用经皮血管成形术技术递送裸质粒DNA进行肺动脉基因转移是可行的。使用裸质粒DNA消除了与辅助剂相关的毒性可能性。所描述的技术为体内研究肺血管生物学和开发未来的基因治疗提供了新方法。
