Minicircle DNA Vector-mediated Augmenter Of Liver Regeneration (ALR) Gene Therapy For Liver Fibrosis In Rats

Liver fibrosis results in cirrhosis,liver cancer and liver failure,which is a major cause of mortality worldwide.Gene therapy is a relatively new paradigm in medicine,with enormous therapeutic potential.The development of an efficient and safe delivery system is essential for clinical gene therapy.Minicircle DNA vectors,which are composed solely of eukaryotic sequences,demonstrate favorable safety profiles,lack immunogenicity,are the most promising non-virus vector used in gene therapy.In the present study,we evaluated the antifibrotic effect of ALR gene therapy mediated by a minicircle vector(MC-hALR)in liver fibrotic rats.Compared with traditional plasimid(pcDNA3.1-hALR),even when we reduced the transfection dose of MC-hALR to 30%(w),the in vitro and in vivo results still demonstrated the higher and more sustained ALR gene expression;the results in liver fibrotic rats that received MC-hALR throμgh Hydrodynamics-based transfection(HBT)for 8 weeks indicated that the minicircle DNA vector produced a more effective gene therapy effect than pcDNA3.1-hALR,even when we reduced the treatment dose of MC-hALR to 30%(w)and the treatment frequency from weekly to biweekly;the in vitro results still demonstrated that the relative overexpression of MC-hALR significantly blocked increases in transforming growth factor-β1(TGF-β1),platelet derived growth factor-BB(PDGF-BB)and α-smooth muscle aorta(α-SMA)levels,effectively suppressed the production of collagens,especially collagen I,effectively alleviates liver injury and fibrosis in rats,thereby improving the survival rate of liver fibrotic rats;it is preliminarily concluded that the relative overexpression of MC-hALR inhibits the activation of hepatic stellate cells(HSCs),thereby alleviating liver fibrosis in rats.To our knowledge,this is the first report on the preparation of MC-hALR and MC-hALR gene therapy by HBT for liver fibrosis.MC-hALR gene delivery maximizes safety,efficacy and is a new approach in the treatment of liver fibrosis in the furture.