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Lentiviral vectors targeting WASp expression to hematopoietic cells, efficiently transduce and correct cells from WAS patients

Gene Therapy volume 14, pages 415–428 (2007)Cite this article

Abstract

Gene therapy has been proposed as a potential treatment for Wiskott–Aldrich syndrome (WAS), a severe primary immune deficiency characterized by multiple hematopoietic-specific cellular defects. In order to develop an optimal lentiviral gene transfer cassette for this application, we compared the performance of several internal promoters in a variety of cell lineages from human WAS patients. Vectors using endogenous promoters derived from short (0.5 kb) or long (1.6 kb) 5′ flanking sequences of the WAS gene, expressed the transgene in T, B, dendritic cells as well as CD34+ progenitor cells, but functioned poorly in non-hematopoietic cells. Defects of T-cell proliferation and interleukin-2 production, and the cytoskeletal anomalies in WAS dendritic cells were also corrected. The levels of reconstitution were comparable to those obtained following transduction with similar lentiviral vectors incorporating constitutive PGK-1, EF1-alpha promoters or the spleen focus forming virus gammaretroviral LTR. Thus, native regulatory sequences target the expression of the therapeutic WAS transgene to the hematopoietic system, as is naturally the case for WAS, and are effective for correction of multiple cellular defects. These vectors may have significant advantages for clinical application in terms of natural gene regulation, and reduction in the potential for adverse mutagenic events.

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Acknowledgements

We thank the patients and their families. We are grateful to A Fischer, C Picard, B Varet (Hôpital Necker, Paris), R Favier (Hôpital Trousseau, Paris) and S Martino (Department of Pediatrics, Turin University) for providing WAS patient samples. We are thankful to Genethon collaborators, in particular to G Griffith, S Martin, S Bucher, S Potier, A Jollet, K Seye and J Blondeau for help with vector construction, production and transduction experiments, to T Larmonier for establishment and maintenance of patient B-LCL lines. Special thanks to D Stockholm for help with image acquisition and analysis. We are grateful to S Tsuchyia (Tohoku University, Sendai, Japan) for providing the 5A5 anti-WASP mAbs and to L Naldini (Tiget, Milan, Italy) for providing the pRRL vector system. This work was supported by equipment funds from ‘Génopole Recherche’, Evry, France, by grants from the AFM (French Muscular Dystrophy Association, Evry, France) and Italian Telethon (to MGR, LD), from the EU CONSERT integrated project (FP6-Project No. 005242), the Wellcome Trust (AJT) and EU STREP EURO-POLICY PID (FP6 Project No. SP23-CT-2005-006411, MPB).

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Author notes

  1. S Charrier, L Dupré and S Scaramuzza: These authors contributed equally to this work.

Authors and Affiliations

  1. INSERM, U790, Evry, France

    S Charrier, L Jeanson-Leh & A Galy

  2. Université Paris 11, Evry, France

    S Charrier, L Jeanson-Leh & A Galy

  3. Généthon, Evry, France

    S Charrier, L Jeanson-Leh & A Galy

  4. San Raffaele Telethon Institute for Gene Therapy, Milan, Italy

    L Dupré, S Scaramuzza, F Cattaneo, A Aiuti & M Grazia Roncarolo

  5. Institute of Child Health, Molecular Immunology Unit, London, UK

    M P Blundell & A J Thrasher

  6. University College, London, UK

    M P Blundell & A J Thrasher

  7. Great Ormond Street Hospital for Children NHS Trust, London, UK

    M P Blundell & A J Thrasher

  8. CNRS UMR8115, Evry, France

    O Danos

  9. Généthon, Evry, France

    O Danos

  10. Genomining, Montrouge, France

    R Eckenberg

  11. Università Vita-Salute San Raffaele, Milan, Italy

    M Grazia Roncarolo

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  1. S Charrier
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  2. L Dupré
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Corresponding author

Correspondence to A Galy.

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Supplementary Information accompanies the paper on Gene Therapy website (http://www.nature.com/gt).

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Charrier, S., Dupré, L., Scaramuzza, S. et al. Lentiviral vectors targeting WASp expression to hematopoietic cells, efficiently transduce and correct cells from WAS patients. Gene Ther 14, 415–428 (2007). https://doi.org/10.1038/sj.gt.3302863

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  • DOI: https://doi.org/10.1038/sj.gt.3302863

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