title: Attenuating Ischemia-Reperfusion Injury in Kidney Transplantation by Perfusing Donor Organs With siRNA Cocktail Solution.
abstract: BACKGROUND
  Ischemia-reperfusion (I/R) injury is the major cause of delayed renal graft function in kidney transplantation. To date, there are no effective therapeutic approaches for preventing I/R injury. We previously reported that treatment of animals with small interference RNA (siRNA) would prevent warm I/R injury in nontransplant models and cold I/R injury in heart transplantation. In the present study, we further explore the feasibility of protecting grafts from extended cold I/R injury as applied to kidney transplantation by downregulating I/R-associated genes using siRNA.
  
  
  METHODS
  Donor kidneys were intra-arterially perfused with siRNA containing solution during donor excision and preserved in siRNA containing solution. The siRNA-treated donor organs were then implanted into syngeneic recipient mice, and the 2 original kidneys were removed from the recipient. The effect of siRNA solution on extended cold I/R injury was determined by assessing renal function, histopathological change, cell apoptosis, and inflammation.
  
  
  RESULTS
  The perfused siRNA solution knocked down the expression of complement 3, RelB, and Fas in the kidney at the mRNA and protein levels. Administration of siRNA solution reduced the levels of blood urea nitrogen and serum creatinine as compared with control groups. The siRNA cocktail decreased cell apoptosis and histopathological changes in the kidney and prolonged graft survival. The siRNA cocktail also reduced the expression of proinflammatory cytokines, IL-6, and TNFα.
  
  
  CONCLUSIONS
  In conclusion, this is the first demonstration that perfusing donor organs with an siRNA cocktail solution can induce gene silencing in the kidney and prevent kidneys from extended cold I/R injury in kidney transplantation, highlighting the promise of the clinical application of siRNA-based therapies in the preservation of donor organs.
authors.affiliation: 1 Departments of Pathology, Surgery, Oncology and Biostatistics, University of Western Ontario, Ontario, Canada. 2 Lawson Health Research Institute, London, Ontario, Canada. 3 London Health Sciences Centre; London, Ontario, Canada. 4 Zhejiang University, Sir Run Run Shaw Hospital, Hangzhou, China.
authors.forename: Xiufen
authors.initials: X
authors.lastname: Zheng
authors.forename: GuoYao
authors.initials: G
authors.lastname: Zang
authors.forename: Jifu
authors.initials: J
authors.lastname: Jiang
authors.forename: Wenqing
authors.initials: W
authors.lastname: He
authors.forename: Nathan J
authors.initials: NJ
authors.lastname: Johnston
authors.forename: Hong
authors.initials: H
authors.lastname: Ling
authors.forename: Ruiqi
authors.initials: R
authors.lastname: Chen
authors.forename: Xusheng
authors.initials: X
authors.lastname: Zhang
authors.forename: Yanling
authors.initials: Y
authors.lastname: Liu
authors.forename: Aaron
authors.initials: A
authors.lastname: Haig
authors.forename: Patrick
authors.initials: P
authors.lastname: Luke
authors.forename: Anthony M
authors.initials: AM
authors.lastname: Jevnikar
authors.forename: Wei-Ping
authors.initials: WP
authors.lastname: Min
mesh_terms: D000818:Animals; D017209:Apoptosis; D050377:Cold Ischemia / Q000009:adverse effects; D003176:Complement C3 / Q000235:genetics / Q000378:metabolism; D005786:Gene Expression Regulation; D018836:Inflammation Mediators / Q000378:metabolism; D015850:Interleukin-6 / Q000235:genetics / Q000378:metabolism; D007668:Kidney / Q000378:metabolism / Q000473:pathology / Q000503:physiopathology / Q000601:surgery*; D016030:Kidney Transplantation / Q000009:adverse effects / Q000379:methods*; D008297:Male; D008810:Mice, Inbred C57BL; D009926:Organ Preservation / Q000379:methods*; D010477:Perfusion / Q000379:methods*; D034741:RNA, Small Interfering / Q000008:administration & dosage* / Q000378:metabolism; D000067492:RNAi Therapeutics / Q000379:methods*; D020127:Recovery of Function; D015427:Reperfusion Injury / Q000235:genetics / Q000378:metabolism / Q000473:pathology / Q000517:prevention & control*; D015398:Signal Transduction; D013997:Time Factors; D051841:Transcription Factor RelB / Q000235:genetics / Q000378:metabolism; D014409:Tumor Necrosis Factor-alpha / Q000235:genetics / Q000378:metabolism; D019014:fas Receptor / Q000235:genetics / Q000378:metabolism
chemical_list: D003176:Complement C3; C491899:Fas protein, mouse; D018836:Inflammation Mediators; D015850:Interleukin-6; D034741:RNA, Small Interfering; C497594:Relb protein, mouse; D014409:Tumor Necrosis Factor-alpha; D019014:fas Receptor; C577686:interleukin-6, mouse; D051841:Transcription Factor RelB