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Lineage- and stage-restricted lentiviral vectors for the gene therapy of chronic granulomatous disease

Gene Therapy volume 18, pages 1087–1097 (2011)Cite this article

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Abstract

Insertional mutagenesis represents a serious adverse effect of gene therapy with integrating vectors. However, although uncontrolled activation of growth-promoting genes in stem cells can predictably lead to oncological processes, this is far less likely if vector transcriptional activity can be restricted to fully differentiated cells. Diseases requiring phenotypic correction only in mature cells offer such an opportunity, provided that lineage/stage-restricted systems can be properly tailored. In this study, we followed this reasoning to design lentiviral vectors for the gene therapy of chronic granulomatous disease (CGD), an immune deficiency due a loss of nicotinamide adenine dinucleotide phosphate (NADPH) oxidase in phagocytes, most often secondary to mutations in gp91phox. Using self-inactivating HIV1-derived vectors as background, we first expressed enhanced green fluorescent protein (eGFP) from a minimal gp91phox promoter, adding various natural or synthetic transcriptional regulatory elements to foster both specificity and potency. The resulting vectors were assessed either by transplantation or by lentiviral transgenesis, searching for combinations conferring strong and specific expression into mature phagocytic cells. The most promising vector was modified to express gp91phox and used to treat CGD mice. High-level restoration of NADPH activity was documented in granulocytes from the treated animals. We propose that this lineage-specific lentiviral vector is a suitable candidate for the gene therapy of CGD.

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Acknowledgements

We thank L Rigonnot, L Khadam, their staff and the mothers at the Maternity ward of Hopital Louise Michel, Evry, France for providing umbilical cord blood samples. We thank J Dick for his support in performing experiments. We also thank L Naldini, S Li, P Salmon and KH Krause for the gifting of reagents, and M Garcia and G Tapia for help with the FACS studies. This work was supported by grants from the Swiss National Science Foundation and from the European Union (FP6 CONSERT and FP7 PERSIST projects).

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Author notes

  1. E Laurenti

    Present address: 5Current addresses: Toronto Medical Discovery Tower, 101 College Street, Toronto, Canada M5G 1L7.,

  2. M Wiznerowicz

    Present address: 6Current addresses: Department of Cancer Diagnostics and Immunology, Greater Poland Cancer Center, Garbary 15, Poznan 61-866, Poland.,

Authors and Affiliations

  1. School of Life Sciences, Laboratory of Virology and Genetics, Ecole Polytechnique Fédérale de Lausanne, Lausanne, Switzerland

    I Barde, E Laurenti, S Verp, M Wiznerowicz, S Offner & D Trono

  2. ‘Frontiers in Genetics’ National Program, Ecole Polytechnique Fédérale de Lausanne, Lausanne, Switzerland

    I Barde, S Verp, M Wiznerowicz, S Offner & D Trono

  3. Généthon, Inserm U951, Evry, France

    A Viornery & A Galy

  4. Division of Cell Biology, German Cancer Research Center and Heidelberg Institute for Stem Cell Technologies and Experimental Medicine, Heidelberg, Germany

    A Trumpp

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Correspondence to D Trono.

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Barde, I., Laurenti, E., Verp, S. et al. Lineage- and stage-restricted lentiviral vectors for the gene therapy of chronic granulomatous disease. Gene Ther 18, 1087–1097 (2011). https://doi.org/10.1038/gt.2011.65

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  • DOI: https://doi.org/10.1038/gt.2011.65

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