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Expression of human Wiskott–Aldrich syndrome protein in patients’ cells leads to partial correction of a phenotypic abnormality of cell surface glycoproteins

Gene Therapy volume 7, pages 314–320 (2000)Cite this article

Abstract

The Wiskott–Aldrich syndrome (WAS) is an uncommon X-linked recessive disease characterized by thrombocytopenia, eczema and immunodeficiency. The biochemical defect of this disorder primarily affects cells derived from bone marrow. To understand better the molecular mechanisms underlying this disease and to evaluate the possibility of correcting the genetic defects in hematopoietic cells, a Moloney murine leukemia virus (MoMLV)- based retroviral vector carrying a functional Wiskott–Aldrich syndrome protein (WASp) cDNA driven by an SV40 promoter (LNS-WASp) was constructed. A packaging cell line containing this vector produced a stable level of WAS protein and maintained a high titer of viral output. Epstein–Barr virus (EBV)-transformed B lymphoblastoid cell lines (B-LCL) from WAS patients, which lack expression of the WAS protein, were transduced by the LNS-WASp retroviral vector and showed expression of WASp by Western blot. Analysis of the O-glycan pattern on cell surface glycoproteins from WAS patients’ B-LCL showed an altered glycosylation pattern, due to increased activity of β-1,6-N-acetylglucosaminyltransferase (C2GnT). Transduction by the retroviral vector carrying the functional WASp cDNA partially restored the abnormal glycosylation pattern, and was accompanied by a decreasing C2GnT activity. These findings imply a functional linkage between the WAS protein and the expression of the glycosyltransferase involved in the O-glycosylation, and also suggest a potential gene therapy via transferring a functional WASp cDNA into hematopoietic cells for Wiskott–Aldrich syndrome.

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Acknowledgements

We would like to thank Mo Dao, Jan Nolta and Bernadette Masinsin for their technical support and Dr Robertson Parkman for his constructive discussions. This work was supported by NIH postdoctoral training grant, (M-MH; No. DK09430–02); NIH grants (DBK; No. R01-DK49000; MF; R37-CA33000); and the Howard Hughes Medical Institute (JMD and UF).

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Author notes

  1. M-M Huang

    Present address: Norris Cancer Center, Rm610, School of Medicine, University of Southern California, Los Angeles, CA 90033, USA

Authors and Affiliations

  1. Division of Research Immunology/Bone Marrow Transplantation, Childrens Hospital Los Angeles, Los Angeles, CA, USA

    M-M Huang, A Wong, X-J Yu, K I Weinberg & D B Kohn

  2. La Jolla Cancer Research Center, The Burnham Institute, La Jolla, CA, USA

    S Tsuboi, M Oh-Eda & M Fukuda

  3. Howard Hughes Medical Institute, Stanford University, School of Medicine, Stanford, CA, USA

    J M Derry & U Francke

  4. Immunex Corp., Seattle, WA 98101, USA

    J M Derry

  5. Department of Genetics, Stanford University, School of Medicine, Stanford, CA, USA

    U Francke

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Huang, MM., Tsuboi, S., Wong, A. et al. Expression of human Wiskott–Aldrich syndrome protein in patients’ cells leads to partial correction of a phenotypic abnormality of cell surface glycoproteins. Gene Ther 7, 314–320 (2000). https://doi.org/10.1038/sj.gt.3301085

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