Transtracheal gene transfection of donor lungs prior to organ procurement increases transgene levels at reperfusion and following transplantation

Stephen D. Cassivi, Jonathan A. Cardella, Stefan Fischer, Mingyao Liu, Arthur S. Slutsky, Shaf Keshavjee

Research output: Contribution to journalArticle

29 Citations (Scopus)

Abstract

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.

Original languageEnglish (US)
Pages (from-to)1181-1188
Number of pages8
JournalJournal of Heart and Lung Transplantation
Volume18
Issue number12
DOIs
StatePublished - Dec 1999
Externally publishedYes

Fingerprint

Tissue and Organ Procurement
Transgenes
Reperfusion
Transfection
Transplantation
Tissue Donors
Lung
Genes
Lung Transplantation
Reperfusion Injury
Transplants
Tissue and Organ Harvesting
Galactosidases

ASJC Scopus subject areas

  • Cardiology and Cardiovascular Medicine
  • Surgery
  • Transplantation

Cite this

Transtracheal gene transfection of donor lungs prior to organ procurement increases transgene levels at reperfusion and following transplantation. / Cassivi, Stephen D.; Cardella, Jonathan A.; Fischer, Stefan; Liu, Mingyao; Slutsky, Arthur S.; Keshavjee, Shaf.

In: Journal of Heart and Lung Transplantation, Vol. 18, No. 12, 12.1999, p. 1181-1188.

Research output: Contribution to journalArticle

Cassivi, Stephen D. ; Cardella, Jonathan A. ; Fischer, Stefan ; Liu, Mingyao ; Slutsky, Arthur S. ; Keshavjee, Shaf. / Transtracheal gene transfection of donor lungs prior to organ procurement increases transgene levels at reperfusion and following transplantation. In: Journal of Heart and Lung Transplantation. 1999 ; Vol. 18, No. 12. pp. 1181-1188.
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title = "Transtracheal gene transfection of donor lungs prior to organ procurement increases transgene levels at reperfusion and following transplantation",
abstract = "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.",
author = "Cassivi, {Stephen D.} and Cardella, {Jonathan A.} and Stefan Fischer and Mingyao Liu and Slutsky, {Arthur S.} and Shaf Keshavjee",
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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

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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