Prolonged transgene expression in glomeruli using an EBV replicon vector system combined with HVJ liposomes

Area of Science:

• Nephrology
• Molecular Biology
• Gene Therapy

Background:

• Gene transfer into kidney glomerular mesangial cells was achieved using hemagglutinating virus of Japan (HVJ) liposomes.
• A key limitation of this method is the transient nature of transgene expression.

Purpose of the Study:

• To achieve long-term gene expression in kidney glomeruli.
• To evaluate the efficacy of Epstein-Barr virus (EBV) replicon-based plasmids for sustained transgene expression in vivo.

Main Methods:

• Utilized an EBV replicon vector (pEBActLuc) containing the minimal EBV components (oriP and EBNA-1) for transgene nuclear retention.
• Delivered the EBV replicon vector and a control plasmid vector (pActLuc) into the kidney via renal artery using artificial viral envelope (AVE)-type HVJ liposomes.
• Quantified glomerular luciferase activity at various time points post-transfection.

Main Results:

• Both pEBActLuc and pActLuc showed equal glomerular luciferase activity on day 4.
• Luciferase gene expression was sustained for at least 56 days in glomeruli transfected with pEBActLuc.
• In contrast, luciferase expression in glomeruli transfected with pActLuc decreased by day 7.

Conclusions:

• The combination of EBV replicon technology and HVJ liposomes facilitates long-term transgene expression in vivo.
• This approach represents a potentially powerful new therapeutic strategy for managing the progression of renal diseases.