Inhibition of HIV-1 infection by lentiviral vectors expressing pol III-promoted anti-HIV RNAs
2003; Elsevier BV; Volume: 8; Issue: 2 Linguagem: Inglês
10.1016/s1525-0016(03)00165-5
ISSN1525-0024
AutoresMing-Jie Li, Gerhard Bauer, Alessandro Michienzi, Jiing-Kuan Yee, Nan-Sook Lee, James Kim, Shirley Xin Li, Daniela Castanotto, John Zaia, John J. Rossi,
Tópico(s)Virus-based gene therapy research
ResumoA 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.
Referência(s)