Receptor-targeted aptamer-siRNA conjugate-directed transcriptional regulation of HIV-1.
Gene-based therapies represent a promising therapeutic paradigm for the treatment of HIV-1, as they have the potential to maintain sustained viral inhibition with reduced treatment interventions. Such an option may represent a long-term treatment alternative to highly active antiretroviral therapy. Methods: We previously described a therapeutic approach, referred to as transcriptional gene silencing (TGS), whereby small noncoding RNAs directly inhibit the transcriptional activity of HIV-1 by targeting sites within the viral promoter, specifically the 5' long terminal repeat (LTR). TGS differs from traditional RNA interference (RNAi) in that it is characterized by concomitant silent-state epigenetic marks on histones and DNA. To deliver TGS-inducing RNAs, we developed functional RNA conjugates based on the previously reported dual function of the gp120 (A-1) aptamer conjugated to 27-mer Dicer-substrate anti-HIV-1 siRNA (dsiRNA), LTR-362. Results: We demonstrate here that high levels of processed guide RNAs localize to the nucleus in infected T lymphoblastoid CEM cell line and primary human CD4+ T-cells. Treatment of the aptamer-siRNA conjugates induced TGS with an ~10-fold suppression of viral p24 levels as measured at day 12 post infection. To explore the silencing efficacy of aptamer-siRNA conjugates in vivo, HIV-1-infected humanized NOD/SCID/IL2 rγnull mice (hu-NSG) were treated with the aptamer-siRNA conjugates. Systemic delivery of the A-1-stick-LTR-362 27-mer siRNA conjugates suppressed HIV-1 infection and protected CD4+ T cell levels in viremia hu-NSG mice. Principle conclusions: Collectively these data suggest that the gp120 aptamer-dsiRNA conjugate design is suitable for systemic delivery of small RNAs that can be used to suppress HIV-1.
['Zhou|Jiehua|J|', 'Lazar|Daniel|D|', 'Li|Haitang|H|', 'Xia|Xin|X|', 'Satheesan|Sangeetha|S|', 'Charlins|Paige|P|', "O'Mealy|Denis|D|", 'Akkina|Ramesh|R|', 'Saayman|Sheena|S|', 'Weinberg|Marc S|MS|', 'Rossi|John J|JJ|', 'Morris|Kevin V|KV|']
D000818:Animals D052157:Aptamers, Nucleotide D001483:Base Sequence D015496:CD4-Positive T-Lymphocytes D045744:Cell Line, Tumor D053487:DEAD-box RNA Helicases D004195:Disease Models, Animal D015967:Gene Expression Regulation, Viral D020868:Gene Silencing D015316:Genetic Therapy D015658:HIV Infections D016325:HIV Long Terminal Repeat D015497:HIV-1 D006801:Humans D051379:Mice D016688:Mice, Inbred NOD D016513:Mice, SCID D008822:Mice, Transgenic D009690:Nucleic Acid Conformation D034741:RNA, Small Interfering D012367:RNA, Viral D043244:Ribonuclease III D014158:Transcription, Genetic
HIV-1 transcriptional gene silencing RNAi gp120 Aptamer