SHARP -2 Gene Silencing by Lentiviral-based Short Hairpin RNA Interference Prolonged Rat Kidney Transplant Recipients' Survival Time

Abstract

Split- and hairy-related protein-2 (SHARP-2) controls the expression of interleukin-2 (IL-2) and interferon-γ (IFN-γ), 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-γ, respectively, compared with a scramble control construct. When donor kidney was perfused with 5 × 10⁷ 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-γ mRNA expression and prolonged rat kidney transplant recipients' survival.

Keywords

Interleukin-2 Interferon-γ Rat kidney transplantation Lentivirus Short hairpin RNA interference Gene silencing

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