Wolstein, OritBoyd, MaureenMillington, MichelleImpey, HelenBoyer, JoshuaHowe, AnnettDelebecque, FredericCornetta, KennethRothe, MichaelBaum, ChristopherNicolson, TamaraKoldej, RachelZhang, JaneKeech, NaomiCamba Colón, JoannaBreton, LouisBartlett, JeffreyAn, Dong SungChen, Irvin SYBurke, BryanSymonds, Geoff P.2016-03-182016-03-182014-02-12Wolstein, O., Boyd, M., Millington, M., Impey, H., Boyer, J., Howe, A., … Symonds, G. P. (2014). Preclinical safety and efficacy of an anti–HIV-1 lentiviral vector containing a short hairpin RNA to CCR5 and the C46 fusion inhibitor. Molecular Therapy. Methods & Clinical Development, 1, 11–. http://doi.org/10.1038/mtm.2013.112329-0501https://hdl.handle.net/1805/8944Gene transfer has therapeutic potential for treating HIV-1 infection by generating cells that are resistant to the virus. We have engineered a novel self-inactivating lentiviral vector, LVsh5/C46, using two viral-entry inhibitors to block early steps of HIV-1 cycle. The LVsh5/C46 vector encodes a short hairpin RNA (shRNA) for downregulation of CCR5, in combination with the HIV-1 fusion inhibitor, C46. We demonstrate here the effective delivery of LVsh5/C46 to human T cell lines, peripheral blood mononuclear cells, primary CD4+ T lymphocytes, and CD34+ hematopoietic stem/progenitor cells (HSPC). CCR5-targeted shRNA (sh5) and C46 peptide were stably expressed in the target cells and were able to effectively protect gene-modified cells against infection with CCR5- and CXCR4-tropic strains of HIV-1. LVsh5/C46 treatment was nontoxic as assessed by cell growth and viability, was noninflammatory, and had no adverse effect on HSPC differentiation. LVsh5/C46 could be produced at a scale sufficient for clinical development and resulted in active viral particles with very low mutagenic potential and the absence of replication-competent lentivirus. Based on these in vitro results, plus additional in vivo safety and efficacy data, LVsh5/C46 is now being tested in a phase 1/2 clinical trial for the treatment of HIV-1 disease.en-USAttribution-NonCommercial-NoDerivs 3.0 United StatesDrug deliveryImmunological disordersTechnologyArticle