Potential mechanisms for cell-based gene therapy to treat HIV/AIDS

Herrera-Carrillo, Elena; Berkhout, Ben

doi:10.1517/14728222.2014.980236

Cited by 13 publications

(8 citation statements)

References 166 publications

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“…Gene therapy represents one alternative approach for treatment of HIV-1 infection (DiGiusto et al, 2013;Herrera-Carrillo & Berkhout, 2015;Tebas et al, 2014). Here, we demonstrated that CRISPR/Cas9 could efficiently cut the CCR5 locus, resulting in various sequence changes and disrupted CCR5 expression.…”

Section: Discussionmentioning

confidence: 99%

Inhibition of HIV-1 infection of primary CD4+ T-cells by gene editing of CCR5 using adenovirus-delivered CRISPR/Cas9

Guan

et al. 2015

Journal of General Virology

191

139

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CCR5 serves as an essential coreceptor for human immunodeficiency virus type 1 (HIV-1) entry, and individuals with a CCR5 D32 variant appear to be healthy, making CCR5 an attractive target for control of HIV-1 infection. The CRISPR/Cas9, which functions as a naturally existing adaptive immune system in prokaryotes, has been recently harnessed as a novel nuclease system for genome editing in mammalian cells. Although CRISPR/Cas9 can be readily delivered into cell lines, due to the large size of the Cas9 protein, efficient delivery of CCR5-targeting CRISPR/Cas9 components into primary cells, including CD4 + T-cells, the primary target for HIV-1 infection in vivo, remains a challenge. In the current study, following design of a panel of top-ranked single-guided RNAs (sgRNAs) targeting the ORF of CCR5, we demonstrate that CRISPR/Cas9 can efficiently mediate the editing of the CCR5 locus in cell lines, resulting in the knockout of CCR5 expression on the cell surface. Next-generation sequencing revealed that various mutations were introduced around the predicted cleavage site of CCR5. For each of the three most effective sgRNAs that we analysed, no significant off-target effects were detected at the 15 top-scoring potential sites. More importantly, by constructing chimeric Ad5F35 adenoviruses carrying CRISPR/Cas9 components, we efficiently transduced primary CD4 + Tlymphocytes and disrupted CCR5 expression, and the positively transduced cells were conferred with HIV-1 resistance. To our knowledge, this is the first study establishing HIV-1 resistance in primary CD4 + T-cells utilizing adenovirus-delivered CRISPR/Cas9.

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Section: Discussionmentioning

confidence: 99%

Inhibition of HIV-1 infection of primary CD4+ T-cells by gene editing of CCR5 using adenovirus-delivered CRISPR/Cas9

Guan

et al. 2015

Journal of General Virology

191

139

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“…7,8 Multiple approaches have been taken for HIV gene therapy, reviewed by Herrera-Carrillo and Berkhout. 9 These strategies include elimination of CCR5 and/or CXCR4 by gene editing using zinc finger 6,10 and CRISPR, 11 gp41 fusion inhibitory peptide maC46, 12 dominant negative viral proteins, [13][14][15] shRNA, 7,8 and chimeric CD4 + T cell receptor (TCR). 16 Although many of these approaches have shown preclinical promise, the growing consensus suggests that a combination gene therapy approach incorporating multiple targets would be required to achieve long-term HIV suppression without ART, and possibly a cure.…”

Section: Introductionmentioning

confidence: 99%

“…16 Although many of these approaches have shown preclinical promise, the growing consensus suggests that a combination gene therapy approach incorporating multiple targets would be required to achieve long-term HIV suppression without ART, and possibly a cure. 9,17 Hence, there is a need for new anti-HIV gene therapy strategies that use novel genes and approaches.…”

Section: Introductionmentioning

confidence: 99%

Conditional Cytotoxic Anti-HIV Gene Therapy for Selectable Cell Modification

Garg

Joshi

2016

Human Gene Therapy

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Gene therapy remains one of the potential strategies to achieve a cure for HIV infection. One of the major limitations of anti-HIV gene therapy concerns recovering an adequate number of modified cells to generate an HIV-proof immune system. Our study addresses this issue by developing a methodology that can mark conditional vector-transformed cells for selection and subsequently target HIV-infected cells for elimination by treatment with ganciclovir (GCV). We used the herpes simplex virus thymidine kinase (TK) mutant SR39, which is highly potent at killing cells at low GCV concentrations. This gene was cloned into a conditional HIV vector, pNL-GFPRRESA, which expresses the gene of interest as well as green fluorescent protein (GFP) in the presence of HIV Tat protein. We show here that TK-SR39 was more potent that wild-type TK (TK-WT) at eliminating infected cells at lower concentrations of GCV. As the vector expresses GFP in the presence of Tat, transient expression of Tat either by Tat RNA transfection or transduction by a nonintegrating lentiviral (NIL) vector marked the cells with GFP for selection. In cells selected by this strategy, TK-SR39 was more potent at limiting virus replication than TK-WT. Finally, in Jurkat cells modified and selected by this approach, infection with CXCR4-tropic Lai virus could be suppressed by treatment with GCV. GCV treatment limited the number of HIV-infected cells, virus production, as well as virus-induced cytopathic effects in this model. We provide proof of principle that TK-SR39 in a conditional HIV vector can provide a safe and effective anti-HIV strategy.

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“…The gene therapy, using various foreign nucleic acids, containing plasmid DNA (pDNA), small interfering RNAs (siRNAs) and oligonucleotides[1–5] to down or unregulated the expression of defective genes, is a promising strategy for treatment of various inherited or acquired diseases, such as AIDS[6, 7], immunodeficiency[8, 9], degenerative disorders[10] and cancer[11, 12]. Gene therapy has been attracted great attention in the field of biomedical sciences and benefited to develop the human health level over the past few decades.…”

Section: Introductionmentioning

confidence: 99%

Synthesis and evaluation of L-arabinose-based cationic glycolipids as effective vectors for pDNA and siRNA in vitro

Guo

Zhang

et al. 2017

PLoS ONE

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Glycolipids might become a new type of promising non-viral gene delivery systems because of their low cytotoxicity, structural diversity, controllable aqua- and lipo-solubility, appropriate density and distribution of positive charges, high transfer efficiency and potential targeting function. In this study, four kinds of L-arabinose-based cationic glycolipids (Ara-DiC12MA, Ara-DiC14MA, Ara-DiC16MA and Ara-DiC18MA) containing quaternary ammonium as hydrophilic headgroup and two alkane chains as hydrophobic domain were synthesized and characterized. They were observed to have strong affinities for plasmid DNA (pDNA) and siRNA, the pDNA can be completely condensed at N/P ratio less than 2, and the siRNA can be completely retarded at N/P ratio less than 3. The dynamic light scattering (DLS) experiment and atomic force microscopy (AFM) experiment demonstrated that cationic lipids and their lipoplexes possessed suitable particle sizes with near-spherical shape and proper ζ-potentials for cell transfection. The Ara-DiC16MA liposome was found to have good transfection efficacy in HEK293, PC-3 and Mat cells compared with other three kinds of liposomes, and also maintain low cytotoxicity and better uptake capability in vitro. Furthermore, the gene silencing assay showed that Ara-DiC14MA and Ara-DiC16MA liposomes have demonstrated effective delivery and higher gene knockdown activity (>80%) in the above mentioned cells than Lipofectamine 2000. These results indicated Ara-DiC16MA can be developed for efficient and low toxic gene delivery.

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Potential mechanisms for cell-based gene therapy to treat HIV/AIDS

Cited by 13 publications

References 166 publications

Inhibition of HIV-1 infection of primary CD4+ T-cells by gene editing of CCR5 using adenovirus-delivered CRISPR/Cas9

Inhibition of HIV-1 infection of primary CD4+ T-cells by gene editing of CCR5 using adenovirus-delivered CRISPR/Cas9

Conditional Cytotoxic Anti-HIV Gene Therapy for Selectable Cell Modification

Synthesis and evaluation of L-arabinose-based cationic glycolipids as effective vectors for pDNA and siRNA in vitro

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