Highly efficient non-viral VEGF gene delivery to human mesenchymal stem cells by lipid based nanoparticles

摘要

Vascular endothelial growth factor(VEGF) is a protein that can trigger blood vessel formation and thus is used for healing cardiovascular diseases, such as myocardial ischemia and bone diseases, such as bone defects. Mesenchymal stem cell-based non-viral VEGF gene therapy is now considered more effective that the direct injection of VEGF protein into diseased sites. In this approach, the VEGF gene carried by a vector such as nanoparticles is transferred into MSCs, followed by transplantation of the transfected MSCs. MSCs produce fresh VEGF protein through gene expression and also differentiate into functional cells such as bone forming cells. However, this method is limited by the low transfection efficiency. The newly formed blood vessels induced by VEGF gene delivery are unstable and require continued VEGF stimulation for about 4 weeks. If VEGF gene expression is lost before this time point, the new blood vessels will regress and disappear. Thus, the sustained expression of VEGF is critical to the success of VEGF gene therapy. To solve these problems, we developed a new human MSCs-based non-viral VEGF gene therapy to achieve highly efficient VEGF gene transfer into hMSC with sustained expression. In our study, we identified an hMSC-targeting peptide using a phage-biopanning technique. we also integrated the hMSC-targeting peptide and VEGF-SB into lipids based nanoparticles to develop a novel non-viral vector for the delivery of the VEGF gene into hMSC with high transfection efficiency and sustained expression. The high transfection efficiency and sustained expression with the lipids based nanoparticles were also verified by flow cytometry and confocal microscopy.