title: Directed HIV-1 evolution of protease inhibitor resistance by second-generation short hairpin RNAs.
abstract: Despite the success of antiretroviral drugs in decreasing AIDS-related mortality, a substantial fraction of HIV-infected patients experience therapy failure due to the emergence of drug-resistant virus variants. For durable inhibition of HIV-1 replication, the emergence of such escape viruses must be controlled. In addition to antiretroviral drugs, RNA interference (RNAi)-based gene therapy can be used to inhibit HIV-1 replication by targeting the viral RNA genome. RNAi is an evolutionary conserved gene silencing mechanism that mediates the sequence-specific breakdown of the targeted mRNA. Here we investigated an alternative strategy combining the activity of a protease inhibitor (PI) with second-generation short hairpin RNAs (shRNAs) designed to specifically block the emergence of PI-resistant HIV-1 variants. We demonstrate that dominant viral escape routes can be effectively blocked by second-generation shRNAs and that virus evolution can be redirected toward less-fit variants. These results are of importance for a deeper understanding of HIV-1 evolution under combined drug and RNAi pressure and may be used to design future therapeutic approaches.
authors: Laboratory of Experimental Virology, Department of Medical Microbiology, Center for Infection and Immunity Amsterdam (CINIMA), Academic Medical Center, University of Amsterdam, Amsterdam, the Netherlands.
authors: Nick C T
authors: NC
authors: Schopman
authors: Anja
authors: A
authors: Braun
authors: Ben
authors: B
authors: Berkhout
mesh_terms: D001483:Base Sequence; D019020:Directed Molecular Evolution; D024882:Drug Resistance, Viral / Q000187:drug effects; D017930:Genes, Reporter; D016679:Genome, Viral; D057809:HEK293 Cells; D015658:HIV Infections / Q000188:drug therapy* / Q000821:virology; D016333:HIV Protease / Q000378:metabolism; D017320:HIV Protease Inhibitors / Q000494:pharmacology*; D015497:HIV-1 / Q000187:drug effects* / Q000235:genetics; D006801:Humans; D016086:Lentivirus; D008156:Luciferases; D008969:Molecular Sequence Data; D010957:Plasmids; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics*; D014162:Transfection; D014779:Virus Replication / Q000187:drug effects / Q000235:genetics
chemical_list: D017320:HIV Protease Inhibitors; D034741:RNA, Small Interfering; D008156:Luciferases; D016333:HIV Protease