A new system for parallel drug screening against multiple-resistant HIV mutants based on lentiviral self-inactivating (SIN) vectors and multi-colour analyses
- Equal contributors
1 Engelhardt-Institute of Molecular Biology, Moscow, Russia
2 Research Dept. Cell and Gene Therapy, Clinic for Stem Cell Transplantation, UCCH, University Medical Center Hamburg-Eppendorf (UKE), Hamburg, Germany
3 Heinrich-Pette-Institute, Leibniz Institute for Experimental Virology, Hamburg, Germany
4 European ScreeningPort, Hamburg, Germany
5 Moscow Institute of Physics and Technology (MIPT), Moscow, Russia
AIDS Research and Therapy 2013, 10:1 doi:10.1186/1742-6405-10-1Published: 3 January 2013
Despite progress in the development of combined antiretroviral therapies (cART), HIV infection remains a significant challenge for human health. Current problems of cART include multi-drug-resistant virus variants, long-term toxicity and enormous treatment costs. Therefore, the identification of novel effective drugs is urgently needed.
We developed a straightforward screening approach for simultaneously evaluating the sensitivity of multiple HIV gag-pol mutants to antiviral drugs in one assay. Our technique is based on multi-colour lentiviral self-inactivating (SIN) LeGO vector technology.
We demonstrated the successful use of this approach for screening compounds against up to four HIV gag-pol variants (wild-type and three mutants) simultaneously. Importantly, the technique was adapted to Biosafety Level 1 conditions by utilising ecotropic pseudotypes. This allowed upscaling to a large-scale screening protocol exploited by pharmaceutical companies in a successful proof-of-concept experiment.
The technology developed here facilitates fast screening for anti-HIV activity of individual agents from large compound libraries. Although drugs targeting gag-pol variants were used here, our approach permits screening compounds that target several different, key cellular and viral functions of the HIV life-cycle. The modular principle of the method also allows the easy exchange of various mutations in HIV sequences. In conclusion, the methodology presented here provides a valuable new approach for the identification of novel anti-HIV drugs.