Successful in vivo and ex vivo transfection of pulmonary artery segments in lung isografts

Byline: Motoki Yano, Carlos H.R. Boasquevisque, Ronald K. Scheule, Mitchell D. Botney, Joel D. Cooper, G.Alexander Patterson Abstract: Objective: Gene transfer to lung grafts may be useful in ameliorating ischemia-reperfusion injury and rejection. Efficient gene transfection to the whole organ may prove problematic. Proximal pulmonary artery endothelial transfection might provide beneficial downstream effects on the whole graft. The aim of this study was to determine the feasibility of transfecting proximal pulmonary artery segments in lung isografts. Methods: Male Fischer rats were divided into six groups. In vivo transfection: In group I (n = 7), a proximal segment of the left pulmonary artery was isolated and injected with saline solution by means of a catheter inserted through the right ventricle. After an exposure period of 20 minutes, clamps were removed and blood flow was restored. In group II (n = 7), the isolated arterial segments were injected with adenovirus carrying the Escherichia coli LacZ gene encoding for [beta]-galactosidase. Ex vivo transfection: In group III (n = 5), arterial segments were injected ex vivo with saline solution and in group IV (n = 5) with the adenovirus construct. In group V (n = 6), arteries were injected with saline solution and in group VI (n = 11) with liposome chloramphenicol acetyl transferase cDNA. In groups I to IV, animals were killed on postoperative day 3 and transgene expression was assessed by Bluo-Gal staining. In groups V and VI, animals were killed on postoperative day 2 and transgene expression was assessed by chloramphenicol acetyl transferase activity assay. Results: Transgene expression was detected grossly and microscopically in endothelial and smooth muscle cells of pulmonary artery segments from all surviving animals of groups II and IV. In group VI, chloramphenicol acetyl transferase activity was significant in all assessed arterial segments. Conclusion: Significant transgene expression is observed in proximal pulmonary artery segments after both in vivo and ex vivo exposure. (J Thorac Cardiovasc Surg 1997;114:793-802) Article History: Received 7 May 1997; Accepted 9 June 1997 Article Note: (footnote) [star] From the Divisions of Cardiothoracic Surgerya and Respiratory and Critical Care Medicine,b Departments of Surgery and Medicine, Washington University School of Medicine, St. Louis, Mo., and Genzyme Corporation,c Framingham, Mass., [star][star] Supported by National Institutes of Health grant 1 R01 HL41281., a Read at the Seventy-seventh Annual Meeting of The American Association for Thoracic Surgery, Washington, D.C., May 4-7, 1997., aa Address for reprints: G. Alexander Patterson, MD, 3108 Queeny Tower, One Barnes Hospital Plaza, St. Louis, MO 63110., acents 0022-5223/97 $5.00 +0 12/6/83785