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Purging of leukemia-contaminated bone marrow grafts using suicide adenoviral vectors: an in vivo murine experimental model

Gene Therapy volume 10, pages 1328–1335 (2003)Cite this article

Abstract

Autologous bone marrow transplantation is an alternative therapeutic option for acute myeloid leukemia patients lacking a compatible donor. However, bone marrow from these patients may contain residual leukemic cells that should be ideally eliminated prior to the infusion of the graft. With the aim of developing more efficient protocols of graft purging, adenoviral-mediated gene transfer protocols have been conducted. We studied whether suicide adenoviral vectors expressing the cytosine deaminase gene (AdCD) could be used for selectively killing leukemic WEHI-3B cells. The AdCD transduction followed by the 5-fluorocytosine exposure abrogated the growth of WEHI-3B cells in vitro, with a minimal effect on normal hematopietic progenitors. To test the efficacy of the purging protocol in vivo, bone marrow cells were mixed with syngenic WEHI-3B cells and this chimeric cell population was transduced with AdCD vectors. Infected cells were injected into myeloablated Balb-c mice, which then received a 5-fluorocytosine treatment for 4 days. All mice transplanted with unpurged bone marrow developed leukemia and died. However, 90% of recipients receiving the purging treatment were healthy up to 9 months post-transplantation and had a perfectly re-established hematopoietic system, without any signal of leukemic cell presence. In conclusion, suicide adenoviral vectors are proposed as a tool for the purging of Adenoviral-susceptible myeloid leukemia cells contaminating autologous bone marrow grafts.

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Acknowledgements

We thank I Ormán for expert assistance with the flow cytometry analysis, and S García, E López and J Martínez for excellent technical assistance. Supported by grants from Comisión Interministerial de Ciencia y Tecnología (CICYT; SAF98-C04-C01). JG-C was supported by grants from Programa de Formación de Personal Investigador (Ref. PN95 05413749) -CICYT- and Instituto de Estudios de la Energía, CIEMAT.

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Authors and Affiliations

  1. Gene Therapy Program, CIEMAT/Fundación Marcelino Botín, Av. Complutense, Madrid, Spain

    J Garcia-Castro, P Rio, J C Segovia & J A Bueren

  2. Centro de Transfusión de la Comunidad de Madrid, Avda de Menéndez Pelayo, Madrid, Spain

    R Lillo & F Garcia-Sanchez

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Garcia-Castro, J., Rio, P., Lillo, R. et al. Purging of leukemia-contaminated bone marrow grafts using suicide adenoviral vectors: an in vivo murine experimental model. Gene Ther 10, 1328–1335 (2003). https://doi.org/10.1038/sj.gt.3301993

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