title: Increasing resistance of tubular epithelial cells to apoptosis by shRNA therapy ameliorates renal ischemia-reperfusion injury.
abstract: Renal tubular epithelial cells (TEC) die by apoptosis or necrosis in renal ischemia-reperfusion injury (IRI). Fas/Fas ligand-dependent fratricide is critical in TEC apoptosis, and Fas promotes renal IRI. Therefore, targeting Fas or caspase-8 may have therapeutic potential for renal injury in kidney transplant or failure. RNA silencing by short hairpin RNA (shRNA) is a novel strategy to down-regulate protein expression. Using this approach, silencing of Fas or caspase-8 by shRNA to prevent TEC apoptosis and IRI was evaluated. IRI was induced by renal artery clamping for 45 or 60 min at 32 degrees C in uninephrectomized C57BL/6 mice. Here, we showed that Fas or pro-caspase-8 expression was significantly knocked down in TEC by stable expression of shRNA, resulting in resistance to apoptosis induced by superoxide, IFN-gamma/TNF-alpha and anti-Fas antibody. Inferior vena cava delivery of pHEX-small interfering RNA targeting Fas or pro-caspase-8 resulted in protection of kidney from IRI, indicated by reduction of renal tubular injury (necrosis and apoptosis) and serum creatinine or blood urea nitrogen. Our data suggest that shRNA-based therapy targeting Fas and caspase-8 in renal cells can lead to protection of kidney from IRI. Attenuation of pro-apoptotic proteins using genetic manipulation strategies such as shRNA might represent a novel strategy to promote kidney allograft survival from rejection or failure.
authors.affiliation: Department of Medicine, The University of Western Ontario, London, Ontario, Canada. dcaigan@uwo.ca
authors.forename: C
authors.initials: C
authors.lastname: Du
authors.forename: S
authors.initials: S
authors.lastname: Wang
authors.forename: H
authors.initials: H
authors.lastname: Diao
authors.forename: Q
authors.initials: Q
authors.lastname: Guan
authors.forename: R
authors.initials: R
authors.lastname: Zhong
authors.forename: A M
authors.initials: AM
authors.lastname: Jevnikar
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000187:drug effects / Q000502:physiology*; D053181:Caspase 8 / Q000187:drug effects; D004195:Disease Models, Animal; D015870:Gene Expression; D016030:Kidney Transplantation / Q000473:pathology*; D007684:Kidney Tubules / Q000098:blood supply*; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D034741:RNA, Small Interfering / Q000235:genetics / Q000494:pharmacology*; D015427:Reperfusion Injury / Q000378:metabolism / Q000473:pathology* / Q000517:prevention & control*; D016896:Treatment Outcome; D019014:fas Receptor / Q000187:drug effects
chemical_list: C491899:Fas protein, mouse; D034741:RNA, Small Interfering; D019014:fas Receptor; D053181:Caspase 8