title: Efficient myocyte gene delivery with complete cardiac surgical isolation in situ.
abstract: BACKGROUND
  Previously, we used cardiopulmonary bypass with incomplete cardiac isolation and antegrade administration of vector for global cardiac gene delivery. Here we present a translatable cardiac surgical procedure that allows for complete surgical isolation of the heart in situ with retrograde (through the coronary venous circulation) administration of both vector and endothelial permeabilizing agents to increase myocyte transduction efficiency.
  
  
  METHODS
  In 6 adult dogs the heart was completely isolated with tourniquets placed around both vena cavae and cannulas and all pulmonary veins. On cardiopulmonary bypass, the aorta and pulmonary artery were crossclamped, and the heart was isolated. Crystalloid cardioplegia at 4 degrees C containing 10(13) particles of adenovirus encoding LacZ and 15 microg of vascular endothelial growth factor was infused retrograde into the coronary sinus and recirculated for a total of 30 minutes. The dogs were then weaned from cardiopulmonary bypass and allowed to recover. With a catheter, 3 control dogs underwent retrograde infusion of the same cocktail without cardiac isolation or cardiopulmonary bypass.
  
  
  RESULTS
  Beta-galactosidase activities in the cardiopulmonary bypass group were several orders of magnitude higher in both the right and left ventricles when compared with those in the control group (P < .05). X-gal staining from the cardiopulmonary bypass group showed unequivocal evidence of myocyte gene expression globally in a significant proportion of cardiac myocytes. No myocyte gene expression was observed in the control group.
  
  
  CONCLUSIONS
  A novel cardiac surgical technique has been developed. This approach with cardiac isolation and retrograde delivery of vector through the coronary sinus results in efficient myocyte transduction in an adult large animal in vivo.
authors.affiliation: Department of Surgery of the University of Pennsylvania Health System, Philadelphia, Pa 19104, USA. cbridges@pahosp.com
authors.forename: Charles R
authors.initials: CR
authors.lastname: Bridges
authors.forename: Kapil
authors.initials: K
authors.lastname: Gopal
authors.forename: David E
authors.initials: DE
authors.lastname: Holt
authors.forename: Charles
authors.initials: C
authors.lastname: Yarnall
authors.forename: Steven
authors.initials: S
authors.lastname: Cole
authors.forename: Rochelle B
authors.initials: RB
authors.lastname: Anderson
authors.forename: Xiaoqing
authors.initials: X
authors.lastname: Yin
authors.forename: Anthony
authors.initials: A
authors.lastname: Nelson
authors.forename: Benjamin W
authors.initials: BW
authors.lastname: Kozyak
authors.forename: Zhonglin
authors.initials: Z
authors.lastname: Wang
authors.forename: James
authors.initials: J
authors.lastname: Lesniewski
authors.forename: Leonard T
authors.initials: LT
authors.lastname: Su
authors.forename: Danielle M
authors.initials: DM
authors.lastname: Thesier
authors.forename: Hari
authors.initials: H
authors.lastname: Sundar
authors.forename: Hansell H
authors.initials: HH
authors.lastname: Stedman
mesh_terms: D000818:Animals; D006348:Cardiac Surgical Procedures / Q000379:methods*; D004285:Dogs; D015316:Genetic Therapy / Q000379:methods*; D032383:Myocytes, Cardiac