title: Serum-stabilized naked caspase-3 siRNA protects autotransplant kidneys in a porcine model.
abstract: The naked small interfering RNA (siRNA) of caspase-3, a key player in ischemia reperfusion injury, was effective in cold preserved and hemoreperfused kidneys, but not autotransplanted kidneys in our porcine models. Here, chemically modified serum stabilized caspase-3 siRNAs were further evaluated. The left kidney was retrieved and infused by University of Wisconsin solution with/without 0.3 mg caspase-3 or negative siRNA into the renal artery for 24-hour cold storage (CS). After an intravenous injection of 0.9 mg siRNA and right-uninephrectomy, the left kidney was autotransplanted for 2 weeks. The effectiveness of caspase-3 siRNA was confirmed by caspase-3 knockdown in the post-CS and/or post-transplant kidneys with reduced apoptosis and inflammation, while the functional caspase-3 siRNA in vivo was proved by detected caspase-3 mRNA degradation intermediates. HMGB1 protein was also decreased in the post-transplanted kidneys; correlated positively with renal IL-1β mRNA, but negatively with serum IL-10 or IL-4. The minimal off-target effects of caspase-3 siRNA were seen with favorable systemic responses. More importantly, renal function, associated with active caspase-3, HMGB1, apoptosis, inflammation, and tubulointerstitial damage, was improved by caspase-3 siRNA. Taken together, the 2-week autotransplanted kidneys were protected when caspase-3 siRNA administrated locally and systemically, which provides important evidence for future clinical trials.
authors.affiliation: Department of Urology, Zhongshan Hospital, Fudan University, Shanghai Key Laboratory of Organ Transplantation, Shanghai, China.
authors.forename: Cheng
authors.initials: C
authors.lastname: Yang
authors.affiliation: Department of Urology, Zhongshan Hospital, Fudan University, Shanghai Key Laboratory of Organ Transplantation, Shanghai, China.
authors.forename: Tian
authors.initials: T
authors.lastname: Zhao
authors.affiliation: Department of Urology, Zhongshan Hospital, Fudan University, Shanghai Key Laboratory of Organ Transplantation, Shanghai, China.
authors.forename: Zitong
authors.initials: Z
authors.lastname: Zhao
authors.affiliation: Department of Urology, Zhongshan Hospital, Fudan University, Shanghai Key Laboratory of Organ Transplantation, Shanghai, China.
authors.forename: Yichen
authors.initials: Y
authors.lastname: Jia
authors.affiliation: Department of Urology, Zhongshan Hospital, Fudan University, Shanghai Key Laboratory of Organ Transplantation, Shanghai, China.
authors.forename: Long
authors.initials: L
authors.lastname: Li
authors.affiliation: Department of Nephrology, Affiliated Hospital of Nantong University, Medical Research Centre, Medical School, University of Nantong, Nantong, China.
authors.forename: Yufang
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: 1] Department of Urology, Zhongshan Hospital, Fudan University, Shanghai Key Laboratory of Organ Transplantation, Shanghai, China [2] Biomedical Research Center, Zhongshan Hospital, Fudan University, Shanghai, China.
authors.forename: Mangen
authors.initials: M
authors.lastname: Song
authors.affiliation: 1] Department of Urology, Zhongshan Hospital, Fudan University, Shanghai Key Laboratory of Organ Transplantation, Shanghai, China [2] Department of Transfusion, Zhongshan Hospital, Fudan University, Shanghai, China.
authors.forename: Ruiming
authors.initials: R
authors.lastname: Rong
authors.affiliation: Department of Urology, Zhongshan Hospital, Fudan University, Shanghai Key Laboratory of Organ Transplantation, Shanghai, China.
authors.forename: Ming
authors.initials: M
authors.lastname: Xu
authors.affiliation: Transplant Group, Department of Infection, Immunity and Inflammation, University of Leicester, Leicester General Hospital, University Hospitals of Leicester, Leicester, UK.
authors.forename: Michael L
authors.initials: ML
authors.lastname: Nicholson
authors.affiliation: 1] Department of Urology, Zhongshan Hospital, Fudan University, Shanghai Key Laboratory of Organ Transplantation, Shanghai, China [2] Qingpu Branch Zhongshan Hospital, Fudan University, Shanghai, China.
authors.forename: Tongyu
authors.initials: T
authors.lastname: Zhu
authors.affiliation: 1] Department of Urology, Zhongshan Hospital, Fudan University, Shanghai Key Laboratory of Organ Transplantation, Shanghai, China [2] Department of Nephrology, Affiliated Hospital of Nantong University, Medical Research Centre, Medical School, University of Nantong, Nantong, China.
authors.forename: Bin
authors.initials: B
authors.lastname: Yang
mesh_terms: D033942:Adaptor Proteins, Vesicular Transport / Q000378:metabolism; D000818:Animals; D017209:Apoptosis / Q000235:genetics; D064592:Autografts / Q000378:metabolism*; D001483:Base Sequence; D001665:Binding Sites; D053148:Caspase 3 / Q000737:chemistry / Q000235:genetics*; D016207:Cytokines / Q000235:genetics / Q000378:metabolism; D005109:Extracellular Matrix / Q000378:metabolism; D006085:Graft Survival / Q000235:genetics*; D024243:HMGB1 Protein / Q000378:metabolism; D007668:Kidney / Q000378:metabolism / Q000473:pathology / Q000503:physiopathology; D007677:Kidney Function Tests; D016030:Kidney Transplantation; D007962:Leukocytes / Q000378:metabolism; D008297:Male; D023421:Models, Animal; D053594:Myeloid Differentiation Factor 88 / Q000378:metabolism; D009195:Peroxidase / Q000378:metabolism; D034622:RNA Interference; D012333:RNA, Messenger / Q000235:genetics; D034741:RNA, Small Interfering / Q000737:chemistry / Q000235:genetics*; D013552:Swine; D051193:Toll-Like Receptors / Q000378:metabolism; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism
chemical_list: D033942:Adaptor Proteins, Vesicular Transport; D016207:Cytokines; D024243:HMGB1 Protein; D053594:Myeloid Differentiation Factor 88; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D051193:Toll-Like Receptors; D014157:Transcription Factors; D009195:Peroxidase; D053148:Caspase 3