SHARP-2 gene silencing by lentiviral-based short hairpin RNA interference prolonged rat kidney transplant recipients' survival time.
Split- and hairy-related protein-2 (SHARP-2) controls the expression of interleukin-2 (IL-2) and interferon-gamma (IFN-gamma), which both play a key role in transplant rejection. This study was designed to investigate whether SHARP-2 short hairpin RNA interference (shRNAi) could prolong the survival of rat kidney transplant recipients. A lentiviral-based shRNAi construct, LV-SHARP-2iC, showed a SHARP-2 gene silencing efficiency of 84% in normal rat kidney cells. In activated T-cells, SHARP-2 gene silencing with the LV-SHARP-2iC construct resulted in 61% and 69% down-regulation of IL-2 and IFN-gamma, respectively, compared with a scramble control construct. When donor kidney was perfused with 5 x 10(7) transforming units of the LV-SHARP-2iC construct, the median survival time of the transplant recipients was prolonged by 4 - 5 days compared with control groups. In conclusion, recombinant lentiviral LV-SHARP-2iC construct effectively silenced SHARP-2 gene expression, which reduced IL-2 and IFN-gamma mRNA expression and prolonged rat kidney transplant recipients' survival.
Shou
Z
Z
Kidney Disease Centre, The First Affiliated Hospital, Zhejiang University, School of Medicine, Hangzhou, China. zfshou@zju.edu.cn
Xiao
H
H
Xu
Y
Y
Wang
Y
Y
Yang
Y
Y
Jiang
H
H
Chen
J
J
Yamada
K
K
Miyamoto
K
K
D000818:Animals; D051792:Basic Helix-Loop-Helix Proteins / Q000235:genetics* / Q000378:metabolism; D002460:Cell Line; D005786:Gene Expression Regulation; D049452:Green Fluorescent Proteins / Q000378:metabolism; D018398:Homeodomain Proteins / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D007371:Interferon-gamma / Q000235:genetics / Q000378:metabolism; D007376:Interleukin-2 / Q000235:genetics / Q000378:metabolism; D053208:Kaplan-Meier Estimate; D016030:Kidney Transplantation; D016086:Lentivirus / Q000235:genetics*; D008213:Lymphocyte Activation / Q000276:immunology; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000378:metabolism*; D051381:Rats; D013601:T-Lymphocytes / Q000378:metabolism; D013997:Time Factors
D051792:Basic Helix-Loop-Helix Proteins; D049452:Green Fluorescent Proteins; D018398:Homeodomain Proteins; D007371:Interferon-gamma; D007376:Interleukin-2; D034741:RNA, Small Interfering; C491538:Bhlhe40 protein, rat