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A Novel Approach to Block HIV-1 Coreceptor CXCR4 in Non-toxic Manner

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Abstract

The chemokine receptor CXCR4 is one of the major coreceptors for human immunodeficiency virus type 1 (HIV-1) and considered as an important therapeutic target. Knockdown of CXCR4 by RNA interference has emerged as a promising strategy for combating HIV-1 infection. However, there is a potential drawback to this strategy as undesired side effects may occur due to the loss of natural function of CXCR4. In this study, we developed a novel approach using a single lentiviral vector to express simultaneously CXCR4 dual-shRNAs and an shRNA-resistant CXCR4 mutant possessing the most possible natural functions of CXCR4 and reduced HIV-1 coreceptor activity. Via this approach we achieved the replacement of endogenous CXCR4 by CXCR4 mutant P191A that could compensate the functional loss of endogenous CXCR4 and significant reduction of HIV-1 replication by 59.2 %. Besides, we demonstrated that construction of recombinant lentiviral vector using 2A peptide-based strategy has significant advantages over using additional promoter-based strategy, including increase of lentivirus titer and avoidance of promoter competition. Therefore, the novel approach to block HIV-1 coreceptor CXCR4 without impairing its normal function provides a new strategy for CXCR4-targeted therapeutics for HIV-1 infection and potential universal applications to knock down a cellular protein in non-toxic manner.

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Acknowledgments

We thank Dr. Qinxue Hu (Wuhan Institute of Virology, Chinese Academy of Sciences) for technical advice and kindly providing HIV-1 proviral DNA pNL4-3, Dr. Luk van Parijs for kindly providing the plasmid pLentilox3.7, WJ Ouyang for providing cell lines, Yan Wang for technical support of flow cytometric analysis, and RD Hao and SQ Xu for assistance in experiments. The following reagents were obtained through the NIH AIDS Research and Reference Reagent Program, Division of AIDS, NIAID, NIH: Ghost X4 cell line from Dr. Vineet N. KewalRamani and Dr. Dan R. Littman, U373-CXCR4 from Dr. Michael Emerman, and pNL4-3.Luc.R-E- from Dr. Nathaniel Landau. This work was funded by the China National Special Research Program of Major Infectious Diseases (Nos. 2012ZX10001006-002 and 2014ZX10001003) and Hubei Provincial Science & Technology Innovation Team Grant (#2012FFA043). D.G. is also supported by Hubei Province’s Outstanding Medical Academic Leader Program.

Conflict of interest

The authors declare no conflict of interest.

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Author notes

  1. Ji-An Pan

    Present address: Department of Molecular Genetics and Microbiology, Stony Brook University, Stony Brook, NY, 11794, USA

Authors and Affiliations

  1. State Key Laboratory of Virology and Modern Virology Research Center, College of Life Sciences, Wuhan University, Wuhan, 430072, People’s Republic of China

    Ye Liu, Jieqiong Zhou, Ji-An Pan, Prudence Mabiala & Deyin Guo

  2. Institute of Medical Virology, Wuhan University School of Medicine, Wuhan, 430072, People’s Republic of China

    Deyin Guo

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Correspondence to Deyin Guo.

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Liu, Y., Zhou, J., Pan, JA. et al. A Novel Approach to Block HIV-1 Coreceptor CXCR4 in Non-toxic Manner. Mol Biotechnol 56, 890–902 (2014). https://doi.org/10.1007/s12033-014-9768-7

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