title: HSP70, Peroxiredoxin-3 and -6 are upregulated during renal warm ischaemia in a donation after circulatory death model.
abstract: BACKGROUND
  The use of donation after circulatory death (DCD) kidneys for transplantation is increasing. Subsequent delayed graft function is related to ischaemia/reperfusion injury (I/R), warm ischaemia (WI) being one of the main contributing factors. This proteomics study aimed to identify candidate biomarkers of WI.
  
  
  METHODS
  Termination biopsies were obtained over 180min in 6 pigs. Proteins were subjected to differential in-gel electrophoresis (DIGE) and identified using LC MS/MS.
  
  
  RESULTS
  Thirty nine protein spots showed significant changes in expression (ANOVA, p<0.05). Peroxiredoxin-3 and -6 (PRX3 and PRX6) were expressed with a fold change (FD) of +1.8 (p=0.03 and 0.02 respectively). A significant upregulation of Alpha-2-HS-glycoprotein (A2HSG, FD+1.9, p=0.047) and heat-shock protein 70-1b (HSP70-1b, FD+2.1 p=0.002) was recorded.
  
  
  CONCLUSIONS
  The expression of PRX3, PRX6 and HSP70-1b during the first 30min of WI may be critical in measuring cellular responses. This is the first large animal model to describe the novel candidate biomarker, structural protein A2HSG. A2HSG upregulation during WI alone in this study is encouraging and further assessment in a DCD auto-transplant model is warranted.
  
  
  CONCLUSIONS
  Warm ischaemia (WI) during donation after circulatory death (DCD) organ retrieval is associated with higher rates of post transplant organ dysfunction. The cellular and molecular mechanism of this paradigm is poorly reported. The work carried out in this large animal study has been performed to enable better understanding of protein expression during DCD WI at the time of retrieval. We have identified differential increased expression of PRX3, PRX6 and HSP70 during the first 30min of WI. Observation of this behaviour has not been reported before. Application of these results in a reperfusion model or autograft animal study would further help study of the named proteins as clinical biomarkers of WI. Alpha 2-HS Glycoprotein (A2HSG) species were also differentially expressed during the WI period. This remains a novel finding. Assessment of A2HSG is also recommended for further study in a reperfusion context. Previous reports of A2HSG have suggested an association in chronic kidney disease and diabetes, but no association with WI has previously been noted in either small or large animals.
authors.affiliation: Department of Renal Transplantation, St. George's Hospital NHS Trust, Blackshaw Rd., Tooting, London SW17 0QT, United Kingdom. Electronic address: mhossain@nhs.net.
authors.forename: Mohammad A
authors.initials: MA
authors.lastname: Hossain
authors.affiliation: Cardiovascular Sciences Research Centre, Division of Clinical Sciences, St. George's, University of London, Cranmer Terrace, Tooting, London SW17 0RE, United Kingdom.
authors.forename: Ayesha I
authors.initials: AI
authors.lastname: De Souza
authors.affiliation: Department of Renal Transplantation, St. George's Hospital NHS Trust, Blackshaw Rd., Tooting, London SW17 0QT, United Kingdom.
authors.forename: Atul
authors.initials: A
authors.lastname: Bagul
authors.affiliation: Department of Renal Transplantation, St. George's Hospital NHS Trust, Blackshaw Rd., Tooting, London SW17 0QT, United Kingdom; Division of Clinical Sciences: Renal Medicine, St. George's, University of London, Cranmer Terrace, Tooting, London SW17 0RE, United Kingdom.
authors.forename: Iain A M
authors.initials: IA
authors.lastname: MacPhee
authors.affiliation: Department of Nephrology and Transplantation, Guy's Hospital, Great Maze Pond, London SE1 9RT, United Kingdom.
authors.forename: Nicos
authors.initials: N
authors.lastname: Kessaris
authors.affiliation: Department of Renal Transplantation, St. George's Hospital NHS Trust, Blackshaw Rd., Tooting, London SW17 0QT, United Kingdom.
authors.forename: Mohamed A
authors.initials: MA
authors.lastname: Morsy
mesh_terms: D000818:Animals; D002853:Chromatography, Liquid; D015180:Electrophoresis, Gel, Two-Dimensional; D018840:HSP70 Heat-Shock Proteins / Q000096:biosynthesis*; D006801:Humans; D007668:Kidney / Q000378:metabolism*; D016030:Kidney Transplantation; D013058:Mass Spectrometry; D008954:Models, Biological; D061105:Peroxiredoxin III / Q000096:biosynthesis*; D054465:Peroxiredoxin VI / Q000096:biosynthesis*; D040901:Proteomics; D013552:Swine; D015854:Up-Regulation; D052096:Warm Ischemia
chemical_list: D018840:HSP70 Heat-Shock Proteins; D061105:Peroxiredoxin III; D054465:Peroxiredoxin VI
keywords: Donation after circulatory death; Experimental model; Renal transplantation; Warm ischaemia