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Published on: May 24, 2012
High efficiency in vitro gene transfer into vascular tissues using a pseudotyped retroviral vector without
1Department of Surgery/Vascular Division, University of Southern California School of Medicine, Los Angeles, CA, USA.
Gene Therapy
|December 22, 1999
Summary
Vascular gene therapy using murine leukemia virus (MuLV) vectors is improved with vesicular stomatitis virus G glycoprotein (VSV-G) pseudotyping. This enhanced retroviral vector achieves higher gene transfer efficiency in human vascular cells and saphenous veins.
Area of Science:
- Molecular Biology
- Gene Therapy
- Virology
Background:
- Murine leukemia virus (MuLV)-derived retroviral vectors show limited success in vascular gene therapy due to low transduction efficiency.
- Improving gene transfer into vascular tissues is crucial for effective vascular gene therapy.
Purpose of the Study:
- To compare the gene transfer efficiency of amphotropic MuLV and vesicular stomatitis virus G glycoprotein (VSV-G)-pseudotyped MuLV vectors in vascular tissues.
- To evaluate the efficacy of VSV-G pseudotyped MuLV for vascular gene therapy applications.
Main Methods:
- Comparison of transduction efficiency of amphotropic MuLV and VSV-G pseudotyped MuLV vectors in human endothelial cells (EC), smooth muscle cells (SMC), and saphenous veins (SV).
- Transduction of the tissue plasminogen activator (tPA) gene into EC using both vectors and measurement of tPA antigen and enzymatic activity.
- Assessment of reverse transcriptase activity and cell proliferation's role in transduction.
Main Results:
- VSV-G pseudotyped MuLV demonstrated significantly higher transduction efficiency (90%) in human EC and SMC compared to amphotropic MuLV (20%).
- A six- to 10-fold greater transduction efficiency was observed in cultured saphenous veins using VSV-G pseudotyped MuLV.
- Transduction with VSV-G pseudotyped MuLV resulted in a 12-fold increase in tPA antigen and a 38-fold increase in tPA enzymatic activity in EC, with no detectable pseudotransduction.
Conclusions:
- MuLV pseudotyped with VSV-G envelope glycoprotein is a highly effective retroviral vector for vascular gene therapy.
- VSV-G pseudotyped MuLV significantly enhances gene transfer efficiency in vascular tissues both in vitro and in vivo.
- The transduction process is dependent on reverse transcription and cell proliferation.
