title: Long-term inhibition of HIV-1 infection in primary hematopoietic cells by lentiviral vector delivery of a triple combination of anti-HIV shRNA, anti-CCR5 ribozyme, and a nucleolar-localizing TAR decoy.
abstract: Combinatorial therapies for the treatment of HIV-1 infection have proven to be effective in reducing patient viral loads and slowing the progression to AIDS. We have developed a series of RNA-based inhibitors for use in a gene therapy-based treatment for HIV-1 infection. The transcriptional units have been inserted into the backbone of a replication-defective lentiviral vector capable of transducing a wide array of cell types, including CD34+ hematopoietic progenitor cells. The combinatorial therapeutic RNA vector harbors a U6 Pol III promoter-driven short hairpin RNA (shRNA) targeting the rev and tat mRNAs of HIV-1, a U6 transcribed nucleolar-localizing TAR RNA decoy, and a VA1-derived Pol III cassette that expresses an anti-CCR5 ribozyme. Each of these therapeutic RNAs targets a different gene product and blocks HIV infection by a distinct mechanism. Our results demonstrate that the combinatorial vector suppresses HIV replication long term in a more-than-additive fashion relative to the single shRNA or double shRNA/ribozyme or decoy combinations. Our data demonstrate the validity and efficacy of a combinatorial RNA-based gene therapy for the treatment of HIV-1 infection.
authors.affiliation: Division of Molecular Biology, Beckman Research Institute of the City of Hope, Duarte, CA 91010, USA.
authors.forename: Ming-Jie
authors.initials: MJ
authors.lastname: Li
authors.forename: James
authors.initials: J
authors.lastname: Kim
authors.forename: Shirley
authors.initials: S
authors.lastname: Li
authors.forename: John
authors.initials: J
authors.lastname: Zaia
authors.forename: Jiing-Kuan
authors.initials: JK
authors.lastname: Yee
authors.forename: Joseph
authors.initials: J
authors.lastname: Anderson
authors.forename: Ramesh
authors.initials: R
authors.lastname: Akkina
authors.forename: John J
authors.initials: JJ
authors.lastname: Rossi
mesh_terms: D002478:Cells, Cultured; D004258:DNA Polymerase III / Q000235:genetics*; D015870:Gene Expression; D018014:Gene Transfer Techniques; D015756:Genes, tat; D005822:Genetic Vectors; D015658:HIV Infections / Q000235:genetics / Q000628:therapy*; D015497:HIV-1 / Q000235:genetics*; D006801:Humans; D016086:Lentivirus / Q000235:genetics*; D008214:Lymphocytes; D011401:Promoter Regions, Genetic; D016337:RNA, Catalytic; D019713:Receptors, CCR5 / Q000627:therapeutic use*; D014161:Transduction, Genetic; D019076:Transgenes
chemical_list: D016337:RNA, Catalytic; D019713:Receptors, CCR5; D004258:DNA Polymerase III