title: Inhibition of HIV-1 infection by lentiviral vectors expressing Pol III-promoted anti-HIV RNAs.
abstract: A primary advantage of lentiviral vectors is their ability to pass through the nuclear envelope into the cell nucleus thereby allowing transduction of nondividing cells. Using HIV-based lentiviral vectors, we delivered an anti-CCR5 ribozyme (CCR5RZ), a nucleolar localizing TAR RNA decoy, or Pol III-expressed siRNA genes into cultured and primary cells. The CCR5RZ is driven by the adenoviral VA1 Pol III promoter, while the human U6 snRNA Pol III-transcribed TAR decoy is embedded in a U16 snoRNA (designated U16TAR), and the siRNAs were expressed from the human U6 Pol III promoter. The transduction efficiencies of these vectors ranged from 96-98% in 293 cells to 15-20% in primary PBMCs. A combination of the CCR5RZ and U16TAR decoy in a single vector backbone gave enhanced protection against HIV-1 challenge in a selective survival assay in both primary T cells and CD34(+)-derived monocytes. The lentiviral vector backbone-expressed siRNAs also showed potent inhibition of p24 expression in PBMCs challenged with HIV-1. Overall our results demonstrate that the lentiviral-based vectors can efficiently deliver single constructs as well as combinations of Pol III therapeutic expression units into primary hematopoietic cells for anti-HIV gene therapy and hold promise for stem or T-cell-based gene therapy for HIV-1 infection.
authors.affiliation: Division of Molecular Biology, Beckman Research Institute of the City of Hope, Duarte, California 91010, USA.
authors.forename: Ming-Jie
authors.initials: MJ
authors.lastname: Li
authors.forename: Gerhard
authors.initials: G
authors.lastname: Bauer
authors.forename: Alessandro
authors.initials: A
authors.lastname: Michienzi
authors.forename: Jiing-Kuan
authors.initials: JK
authors.lastname: Yee
authors.forename: Nan-Sook
authors.initials: NS
authors.lastname: Lee
authors.forename: James
authors.initials: J
authors.lastname: Kim
authors.forename: Shirley
authors.initials: S
authors.lastname: Li
authors.forename: Daniela
authors.initials: D
authors.lastname: Castanotto
authors.forename: John
authors.initials: J
authors.lastname: Zaia
authors.forename: John J
authors.initials: JJ
authors.lastname: Rossi
mesh_terms: D001483:Base Sequence; D002460:Cell Line; D002470:Cell Survival; D002478:Cells, Cultured; D004258:DNA Polymerase III / Q000235:genetics / Q000378:metabolism*; D015870:Gene Expression; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors / Q000235:genetics*; D015658:HIV Infections / Q000235:genetics* / Q000628:therapy*; D016325:HIV Long Terminal Repeat / Q000235:genetics; D006801:Humans; D016086:Lentivirus / Q000235:genetics*; D011401:Promoter Regions, Genetic / Q000235:genetics; D016337:RNA, Catalytic / Q000235:genetics / Q000378:metabolism; D034741:RNA, Small Interfering / Q000235:genetics / Q000378:metabolism*; D019713:Receptors, CCR5 / Q000096:biosynthesis / Q000235:genetics; D013601:T-Lymphocytes / Q000378:metabolism / Q000821:virology
chemical_list: D016337:RNA, Catalytic; D034741:RNA, Small Interfering; D019713:Receptors, CCR5; D004258:DNA Polymerase III