TY - JOUR
T1 - Transtracheal gene transfection of donor lungs prior to organ procurement increases transgene levels at reperfusion and following transplantation
AU - Cassivi, Stephen D.
AU - Cardella, Jonathan A.
AU - Fischer, Stefan
AU - Liu, Mingyao
AU - Slutsky, Arthur S.
AU - Keshavjee, Shaf
N1 - Funding Information:
This work is supported by a grant from the National Sanitarium Association of Canada. Dr. Liu is a scholar of the Medical Research Council of Canada. Dr. Boehler is a recipient of grants from the Swiss National Scientific Foundation and the Swiss Respiratory Society. We also acknowledge the technical support of John Mates, DVM, as well as Johnson & Johnson for providing suture material.
Copyright:
Copyright 2007 Elsevier B.V., All rights reserved.
PY - 1999/12
Y1 - 1999/12
N2 - Background: Gene therapy's potential to modify donor organs to better withstand the process of transplantation has yet to be realized. To determine whether gene transfection is feasible to treat the early post-transplant injury of ischemia-reperfusion, we compared transfection of lungs in the donor prior to organ procurement with transfection of harvested ex vivo lungs in a rat single lung transplant model.MethodsLewis rats (donor transfection [DT]; n = 4) underwent transtracheal adenoviral-mediated transfection with 109 plaque forming unit of the β-galactosidase reporter gene. Donor lungs were harvested following 6 hours of in vivo post-transfection ventilation, and then preserved for 6 hours at 4°C prior to left single-lung transplantation. Ex vivo transfection was performed following organ retrieval; lungs were then preserved at 4°C for 6 hours (EVT6h; n = 6) and 12 hours (EVT12h; n = 6) prior to transplantation. Lung transgene expression was measured by chemiluminescence at reperfusion, and at 2 hours following lung transplantation.ResultsDonor transfection lungs showed significantly higher levels of transgene expression as compared with EVT lungs at the time of reperfusion (DT = 3,408 ± 1,301 relative light units/mg protein; EVT6h = 218 ± 7; EVT12h = 213 ± 26; p < 0.02) and at 2 hours after lung transplantation (DT = 2900 ± 870; EVT6h = 62 ± 27; EVT12h = 123 ± 21; p < 0.005). Transgene expression measured in the heart, liver, kidney, and serum from DT rats demonstrated virtually no evidence of collateral transfection at 12 hours post-transfection (all <5.0).ConclusionsGene transfection of donor lungs produces significantly higher levels of transgene expression in lungs at the critical time of reperfusion and in the early period following lung transplantation as compared to ex vivo transfection of cold preserved lungs. Transtracheal donor-lung transfection does not appear to result in collateral transfection of other transplantable organs. Local adenoviral-mediated transfection of the lungs is possible in the multiorgan donor prior to organ procurement and may provide the optimal strategy for gene therapeutic manipulations to address post-transplant ischemia-reperfusion injury. Copyright (C) 1999 International Society for Heart and Lung Transplantation.
AB - Background: Gene therapy's potential to modify donor organs to better withstand the process of transplantation has yet to be realized. To determine whether gene transfection is feasible to treat the early post-transplant injury of ischemia-reperfusion, we compared transfection of lungs in the donor prior to organ procurement with transfection of harvested ex vivo lungs in a rat single lung transplant model.MethodsLewis rats (donor transfection [DT]; n = 4) underwent transtracheal adenoviral-mediated transfection with 109 plaque forming unit of the β-galactosidase reporter gene. Donor lungs were harvested following 6 hours of in vivo post-transfection ventilation, and then preserved for 6 hours at 4°C prior to left single-lung transplantation. Ex vivo transfection was performed following organ retrieval; lungs were then preserved at 4°C for 6 hours (EVT6h; n = 6) and 12 hours (EVT12h; n = 6) prior to transplantation. Lung transgene expression was measured by chemiluminescence at reperfusion, and at 2 hours following lung transplantation.ResultsDonor transfection lungs showed significantly higher levels of transgene expression as compared with EVT lungs at the time of reperfusion (DT = 3,408 ± 1,301 relative light units/mg protein; EVT6h = 218 ± 7; EVT12h = 213 ± 26; p < 0.02) and at 2 hours after lung transplantation (DT = 2900 ± 870; EVT6h = 62 ± 27; EVT12h = 123 ± 21; p < 0.005). Transgene expression measured in the heart, liver, kidney, and serum from DT rats demonstrated virtually no evidence of collateral transfection at 12 hours post-transfection (all <5.0).ConclusionsGene transfection of donor lungs produces significantly higher levels of transgene expression in lungs at the critical time of reperfusion and in the early period following lung transplantation as compared to ex vivo transfection of cold preserved lungs. Transtracheal donor-lung transfection does not appear to result in collateral transfection of other transplantable organs. Local adenoviral-mediated transfection of the lungs is possible in the multiorgan donor prior to organ procurement and may provide the optimal strategy for gene therapeutic manipulations to address post-transplant ischemia-reperfusion injury. Copyright (C) 1999 International Society for Heart and Lung Transplantation.
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U2 - 10.1016/S1053-2498(99)00095-9
DO - 10.1016/S1053-2498(99)00095-9
M3 - Article
C2 - 10612376
AN - SCOPUS:0032715632
SN - 1053-2498
VL - 18
SP - 1181
EP - 1188
JO - Journal of Heart and Lung Transplantation
JF - Journal of Heart and Lung Transplantation
IS - 12
ER -