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Lentiviral vectors transcriptionally targeted to hematopoietic cells by WASP gene proximal promoter sequences

Gene Therapy volume 12, pages 715–723 (2005)Cite this article

Abstract

The development of vectors that express a therapeutic transgene efficiently and specifically in hematopoietic cells (HCs) is an important goal for gene therapy of hematological disorders. In order to achieve this, we used a 500 bp fragment from the proximal WASP gene promoter to drive the expression of the WASP cDNA in the context of a self-inactivating lentiviral vector. Single-round transduction of WASp-deficient herpesvirus saimiri (HVS)-immortalized cells as well as primary allospecific T cells from Wiskott–Aldrich syndrome (WAS) patients with this vector (WW) resulted in expression levels similar to those of control cells. Non-HCs were transduced with similar efficiency, but the levels of WASp were 135–350 times lower than those achieved in HCs. Additionally, transduction of WASp-deficient cells with WW conferred a selective growth advantage in vitro. Therefore, lentiviral vectors incorporating proximal promoter sequences from the WASP gene confer hematopoietic-specific, and physiological protein expression.

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Acknowledgements

We are indebted to Didier Trono (University of Geneva, Geneva, Switzerland) for providing the HIV packaging pCMVΔR8.91 and envelope pMD.G plasmids and to Nuria Matamoros (Hospital Son Dureta, Palma de Mallorca, Spain) for WAS patient blood. We acknowledge the generous supply of rIL-2 (Hoffman-LaRoche, Nutley, NJ, USA) provided by the National Institutes of Health AIDS reference and reagent program (Rockville, MD, USA). This work was supported by EU Contract Grant number QLT-1999-01090 (WASPNEST; AJT, partner 1; IJM, partner 7 and MS, partner 8); SAF2003-09807-C02-01 (MS) and 02 (IJM); FIS Grant FIS01/3143 and SAF2003-00645 to FM. We also thank Wellcome Trust for financial support to AJT. MGT is a FPU predoctoral fellow from the Spanish Ministry of Education and Culture. GKS is a fellow from the Spanish Agency for International Cooperation (AECI).

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Authors and Affiliations

  1. IPB ‘López Neyra’ CSIC, Granada, Spain

    F Martín & C Frecha

  2. Immunology Unit and Institute of Biopathology and Regenerative Medicine, University of Granada School of Medicine, Granada, Spain

    M Ga Toscano, G K Srivastava & I J Molina

  3. Molecular Immunology Unit, Institute of Child Health, University College London, London, UK

    M Blundell & A J Thrasher

  4. Department of Cell Biology, Physiology and Immunology, University of Córdoba School of Medicine, Córdoba, Spain

    M Santamaría

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Martín, F., Toscano, M., Blundell, M. et al. Lentiviral vectors transcriptionally targeted to hematopoietic cells by WASP gene proximal promoter sequences. Gene Ther 12, 715–723 (2005). https://doi.org/10.1038/sj.gt.3302457

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