title: BacMam Delivery of a Protective Gene to Reduce Renal Ischemia-Reperfusion Injury.
abstract: Ischemia-reperfusion (I/R) injury remains the primary contributor to delayed graft function in kidney transplantation. The beneficial application of manganese superoxide dismutase (sod), delivered by a BacMam vector, against renal I/R injury has not been evaluated previously. Therefore, this study overexpressed sod-2 in proximal tubular epithelial (HK-2) cells and porcine kidney organs during simulated renal I/R injury. Incubation of HK-2 cells with antimycin A and 2-deoxyglucose resulted in a significant decrease in intracellular adenosine triphosphate (ATP) levels; following reperfusion, ATP levels significantly increased over time in cells overexpressing sod-2. In addition, lactate dehydrogenase (LDH) release declined over 72 h in BacMam-transduced injured cells. Ex vivo delivery of sod-2 significantly increased ATP levels in organs after 24 h of cold perfusion. In vitro and ex vivo results suggested that BacMam transduction successfully delivered sod-2, which reduced injury associated with I/R, by improving ATP cell content and decreasing LDH release with a subsequent increase in kidney tissue viability. These data provide further evidence for the potential application of BacMam as a gene delivery system for attenuating injury after cold preservation.
authors.affiliation: 1 Department of Biological and Medical Sciences, Faculty of Health and Life Sciences, Oxford Brookes University , Oxford, United Kingdom.
authors.forename: Elisabetta
authors.initials: E
authors.lastname: Hitchman
authors.affiliation: 2 Oxford Expression Technologies Ltd. , BioInnovation Hub, Oxford, United Kingdom .
authors.forename: Richard B
authors.initials: RB
authors.lastname: Hitchman
authors.affiliation: 1 Department of Biological and Medical Sciences, Faculty of Health and Life Sciences, Oxford Brookes University , Oxford, United Kingdom.
authors.forename: Linda A
authors.initials: LA
authors.lastname: King
mesh_terms: D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D002460:Cell Line; D005822:Genetic Vectors / Q000235:genetics*; D007511:Ischemia / Q000235:genetics / Q000378:metabolism / Q000628:therapy*; D007668:Kidney / Q000378:metabolism; D016030:Kidney Transplantation / Q000379:methods; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D015427:Reperfusion Injury / Q000235:genetics* / Q000378:metabolism / Q000628:therapy*; D061987:Sf9 Cells; D013482:Superoxide Dismutase / Q000378:metabolism; D013552:Swine; D000099156:Superoxide Dismutase 2
chemical_list: D000255:Adenosine Triphosphate; D007770:L-Lactate Dehydrogenase; D013482:Superoxide Dismutase; D000099156:Superoxide Dismutase 2
keywords: ATP; BacMam; Baculovirus; HK-2; SOD; ischemia