title: Safety and Efficacy of Ex Vivo Donor Lung Adenoviral IL-10 Gene Therapy in a Large Animal Lung Transplant Survival Model.
abstract: Ex vivo normothermic lung perfusion (EVLP) is a novel platform and method developed to facilitate functional assessment and implementation of advanced therapies for donor lungs prior to transplantation. This study aimed to determine the safety and immunological and functional benefits of ex vivo adenoviral human interleukin-10 (AdhIL-10) gene delivery to prevent the development of primary graft dysfunction in a large animal survival model. Pig donor lungs were retrieved, preserved for 6 h at 4°C, and then randomly assigned to four groups: (1) AdhIL-10 gene therapy: 12 h EVLP + AdhIL-10 intra-bronchial delivery; (2) EVLP-control: 12 h EVLP; (3) Vector-control: 12 h EVLP + adenoviral vector intra-bronchial delivery; and (4) prolonged hypothermic preservation: additional 12 h of cold ischemia. The left lung was then transplanted and evaluated. The recipients were recovered and kept alive until day 7 post-transplant under standard triple immunosuppression. Plasma levels of hIL-10 were detected in the treatment group throughout the 7 days. Analysis of peripheral blood obtained after transplant showed no signs of hematological, renal, or hepatic toxicity in the AdhIL-10 group. The immediate post-transplant lung function was significantly better in the EVLP-control and AdhIL-10 groups. Gas exchange at day 7 was superior in allografts from the AdhIL-10 group, and the histologic inflammation score was significantly lower. Lymphocytes from AdhIL-10 group harvested from mediastinal lymph nodes at day 7 post-transplantation and co-cultured with donor lymphocytes showed significantly less interferon gamma production in an Enzyme-Linked ImmunoSpot assay when compared with non-treatment groups. It has been demonstrated in this preclinical large animal survival study that ex vivo treatment with AdhIL-10 is safe and improves post-transplant lung function over EVLP alone. Improved function and an immunological advantage in both the innate and adaptive immune responses have been demonstrated.
authors.affiliation: 1 Latner Thoracic Surgery Research Laboratories, Toronto General Research Institute, University Health Network , Toronto, Ontario, Canada.|2 Toronto Lung Transplant Program, University Health Network , Toronto, Ontario, Canada.
authors.forename: Tiago N
authors.initials: TN
authors.lastname: Machuca
authors.affiliation: 1 Latner Thoracic Surgery Research Laboratories, Toronto General Research Institute, University Health Network , Toronto, Ontario, Canada.|2 Toronto Lung Transplant Program, University Health Network , Toronto, Ontario, Canada.
authors.forename: Marcelo
authors.initials: M
authors.lastname: Cypel
authors.affiliation: 1 Latner Thoracic Surgery Research Laboratories, Toronto General Research Institute, University Health Network , Toronto, Ontario, Canada.
authors.forename: Riccardo
authors.initials: R
authors.lastname: Bonato
authors.affiliation: 1 Latner Thoracic Surgery Research Laboratories, Toronto General Research Institute, University Health Network , Toronto, Ontario, Canada.|2 Toronto Lung Transplant Program, University Health Network , Toronto, Ontario, Canada.
authors.forename: Jonathan C
authors.initials: JC
authors.lastname: Yeung
authors.affiliation: 1 Latner Thoracic Surgery Research Laboratories, Toronto General Research Institute, University Health Network , Toronto, Ontario, Canada.
authors.forename: Yi-Min
authors.initials: YM
authors.lastname: Chun
authors.affiliation: 1 Latner Thoracic Surgery Research Laboratories, Toronto General Research Institute, University Health Network , Toronto, Ontario, Canada.|2 Toronto Lung Transplant Program, University Health Network , Toronto, Ontario, Canada.
authors.forename: Stephen
authors.initials: S
authors.lastname: Juvet
authors.affiliation: 1 Latner Thoracic Surgery Research Laboratories, Toronto General Research Institute, University Health Network , Toronto, Ontario, Canada.
authors.forename: Zehong
authors.initials: Z
authors.lastname: Guan
authors.affiliation: 1 Latner Thoracic Surgery Research Laboratories, Toronto General Research Institute, University Health Network , Toronto, Ontario, Canada.|2 Toronto Lung Transplant Program, University Health Network , Toronto, Ontario, Canada.
authors.forename: David M
authors.initials: DM
authors.lastname: Hwang
authors.affiliation: 1 Latner Thoracic Surgery Research Laboratories, Toronto General Research Institute, University Health Network , Toronto, Ontario, Canada.
authors.forename: Manyin
authors.initials: M
authors.lastname: Chen
authors.affiliation: 1 Latner Thoracic Surgery Research Laboratories, Toronto General Research Institute, University Health Network , Toronto, Ontario, Canada.|2 Toronto Lung Transplant Program, University Health Network , Toronto, Ontario, Canada.
authors.forename: Tomohito
authors.initials: T
authors.lastname: Saito
authors.affiliation: 1 Latner Thoracic Surgery Research Laboratories, Toronto General Research Institute, University Health Network , Toronto, Ontario, Canada.
authors.forename: Constantine
authors.initials: C
authors.lastname: Harmantas
authors.affiliation: 3 University of Pennsylvania , Philadelphia, Pennsylvania.
authors.forename: Beverly L
authors.initials: BL
authors.lastname: Davidson
authors.affiliation: 1 Latner Thoracic Surgery Research Laboratories, Toronto General Research Institute, University Health Network , Toronto, Ontario, Canada.|2 Toronto Lung Transplant Program, University Health Network , Toronto, Ontario, Canada.
authors.forename: Thomas K
authors.initials: TK
authors.lastname: Waddell
authors.affiliation: 1 Latner Thoracic Surgery Research Laboratories, Toronto General Research Institute, University Health Network , Toronto, Ontario, Canada.
authors.forename: Mingyao
authors.initials: M
authors.lastname: Liu
authors.affiliation: 1 Latner Thoracic Surgery Research Laboratories, Toronto General Research Institute, University Health Network , Toronto, Ontario, Canada.|2 Toronto Lung Transplant Program, University Health Network , Toronto, Ontario, Canada.
authors.forename: Shaf
authors.initials: S
authors.lastname: Keshavjee
mesh_terms: D000256:Adenoviridae / Q000235:genetics*; D000818:Animals; D004195:Disease Models, Animal; D018014:Gene Transfer Techniques; D015316:Genetic Therapy / Q000379:methods; D005822:Genetic Vectors / Q000235:genetics*; D006085:Graft Survival / Q000235:genetics*; D006801:Humans; D016753:Interleukin-10 / Q000235:genetics*; D008168:Lung / Q000378:metabolism*; D016040:Lung Transplantation / Q000379:methods; D008297:Male; D010477:Perfusion / Q000379:methods; D015427:Reperfusion Injury / Q000235:genetics*; D013552:Swine; D014019:Tissue Donors
chemical_list: D016753:Interleukin-10
keywords: cell-based therapies; disease models—lung; gene regulation; vectors—adenovirus and other DNA viruses; vectors—delivery methods