Abstract
The highly specific mechanism of RNA (RNAi) that inhibits the expression of disease genes is increasingly being harnessed to develop a new class of therapeutics for a wide variety of human maladies. The successful use of small interfering RNAs (siRNAs) for therapeutic purposes requires safe and efficient delivery to specific cells and tissues. Herein, we demonstrate novel cell type-specific dual inhibitory function anti-gp120 aptamer-siRNA delivery systems for HIV-1 therapy, in which both the aptamer and the siRNA portions have potent anti-HIV activities. The envelope glycoprotein is expressed on the surface of HIV-1 infected cells, allowing binding and internalization of the aptamer-siRNA chimeric molecules. The Dicer substrate siRNA delivered by the aptamers is functionally processed by Dicer, resulting in specific inhibition of HIV-1 replication and infectivity in cultured CEM T-cells and primary blood mononuclear cells. Our results provide a set of novel aptamer-targeted RNAi therapeutics to combat HIV and further validate the use of anti-gp120 aptamers for delivery of Dicer substrate siRNAs.
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References
Fire, A., Xu, S., Montgomery, M. K., Kostas, S. A., Driver, S. E., and Mello, C. C. (1998) Potent and specific genetic interference by double-stranded RNA in Caenorhabditis elegans. Nature 391, 806–11.
Zamore, P. D., Tuschl, T., Sharp, P. A., and Bartel, D. P. (2000) RNAi: double-stranded RNA directs the ATP-dependent cleavage of mRNA at 21 to 23 nucleotide intervals. Cell 101, 25–33.
Castanotto, D., and Rossi, J. J. (2009) The promises and pitfalls of RNA-interference-based therapeutics. Nature 457, 426–33.
Kim, D. H., and Rossi, J. J. (2007) Strategies for silencing human disease using RNA interference. Nat Rev Genet 8, 173–84.
de Fougerolles, A., Vornlocher, H. P., Maraganore, J., and Lieberman, J. (2007) Interfering with disease: a progress report on siRNA-based therapeutics. Nat Rev Drug Discov 6, 443–53.
Song, E., Lee, S. K., Wang, J., Ince, N., Ouyang, N., Min, J., Chen, J., Shankar, P., and Lieberman, J. (2003) RNA interference targeting Fas protects mice from fulminant hepatitis. Nat Med 9, 347–51.
Scherer, L., Rossi, J. J., and Weinberg, M. S. (2007) Progress and prospects: RNA-based therapies for treatment of HIV infection. Gene Ther 14, 1057–64.
Martinez, M. A. (2009) Progress in the therapeutic applications of siRNAs against HIV-1. Methods Mol Biol 487, 343–68.
Tsygankov, A. Y. (2009) Current developments in anti-HIV/AIDS gene therapy. Curr Opin Investig Drugs 10, 137–49.
Podlekareva, D., Mocroft, A., Dragsted, U. B., Ledergerber, B., Beniowski, M., Lazzarin, A., Weber, J., Clumeck, N., Vetter, N., Phillips, A., and Lundgren, J. D. (2006) Factors associated with the development of opportunistic infections in HIV-1-infected adults with high CD4+ cell counts: a EuroSIDA study. J Infect Dis 194, 633–41.
Kilby, J. M., and Eron, J. J. (2003) Novel therapies based on mechanisms of HIV-1 cell entry. N Engl J Med 348, 2228–38.
Kwong, P. D., Wyatt, R., Robinson, J., Sweet, R. W., Sodroski, J., and Hendrickson, W. A. (1998) Structure of an HIV gp120 envelope glycoprotein in complex with the CD4 receptor and a neutralizing human antibody. Nature 393, 648–59.
Wyatt, R., and Sodroski, J. (1998) The HIV-1 envelope glycoproteins: fusogens, antigens, and immunogens. Science 280, 1884–8.
Zhou, J., Li, H., Li, S., Zaia, J., and Rossi, J. J. (2008) Novel dual inhibitory function aptamer-siRNA delivery system for HIV-1 therapy. Mol Ther 16, 1481–9.
Zhou, J., Swiderski, P., Li, H., Zhang, J., Neff, C. P., Akkina, R., and Rossi, J. J. (2009) Selection, characterization and application of new RNA HIV gp 120 aptamers for facile delivery of Dicer substrate siRNAs into HIV infected cells. Nucleic Acids Res 37, 3094–109.
Weiss, C. D., and White, J. M. (1993) Characterization of stable Chinese hamster ovary cells expressing wild-type, secreted, and glycosylphosphatidylinositol-anchored human immunodeficiency virus type 1 envelope glycoprotein. J Virol 67, 7060–6.
Vodicka, M. A., Goh, W. C., Wu, L. I., Rogel, M. E., Bartz, S. R., Schweickart, V. L., Raport, C. J., and Emerman, M. (1997) Indicator cell lines for detection of primary strains of human and simian immunodeficiency viruses. Virology 233, 193–8.
Acknowledgments
This work was supported by grants from the National Institutes of Health AI29329 and HL07470 awarded to J.J.R. The HIV-1Ba-L gp120 protein, HIV-1 Bal virus, and CHO-EE and CHO-WT gp160 cell lines (16, 17) were obtained through the NIH AIDS Research and Reference Reagent Program, Division of AIDS, NIAID, NIH.
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© 2011 Humana Press
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Zhou, J., Rossi, J.J. (2011). Aptamer-Targeted RNAi for HIV-1 Therapy. In: van Rij, R. (eds) Antiviral RNAi. Methods in Molecular Biology, vol 721. Humana Press. https://doi.org/10.1007/978-1-61779-037-9_22
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DOI: https://doi.org/10.1007/978-1-61779-037-9_22
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