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In vivo selection of autologous MGMT gene-modified cells following reduced-intensity conditioning with BCNU and temozolomide in the dog model

Cancer Gene Therapy volume 19, pages 523–529 (2012)Cite this article

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Abstract

Chemotherapy with 1,3-bis (2-chloroethyl)-1-nitrosourea (BCNU) and temozolomide (TMZ) is commonly used for the treatment of glioblastoma multiforme (GBM) and other cancers. In preparation for a clinical gene therapy study in patients with glioblastoma, we wished to study whether these reagents could be used as a reduced-intensity conditioning regimen for autologous transplantation of gene-modified cells. We used an MGMT(P140K)-expressing lentivirus vector to modify dog CD34+ cells and tested in four dogs whether these autologous cells engraft and provide chemoprotection after transplantation. Treatment with O6-benzylguanine (O6BG)/TMZ after transplantation resulted in gene marking levels up to 75%, without significant hematopoietic cytopenia, which is consistent with hematopoietic chemoprotection. Retrovirus integration analysis showed that multiple clones contribute to hematopoiesis. These studies demonstrate the ability to achieve stable engraftment of MGMT(P140K)-modified autologous hematopoietic stem cells (HSCs) after a novel reduced-intensity conditioning protocol using a combination of BCNU and TMZ. Furthermore, we show that MGMT(P140K)-HSC engraftment provides chemoprotection during TMZ dose escalation. Clinically, chemoconditioning with BCNU and TMZ should facilitate engraftment of MGMT(P140K)-modified cells while providing antitumor activity for patients with poor prognosis glioblastoma or alkylating agent-sensitive tumors, thereby supporting dose-intensified chemotherapy regimens.

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Acknowledgements

We thank Helen Crawford, Laura Farren and Bonnie Larson for help in preparing this manuscript. We also thank Michele Spector and the Fred Hutchinson Cancer Research Center Dog Lab staff for the care of the dogs. This work was supported in part by National Institutes of Health (Bethesda, MD) grants P01HL036444, R01HL074162 and P30DK056465. H-PK is a Markey Molecular Medicine Investigator and the recipient of the José Carreras/ED Thomas Endowed Chair for Cancer Research. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health nor its subsidiary Institutes and Centers.

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  1. Clinical Research Division, Fred Hutchinson Cancer Research Center, Seattle, WA, USA

    J L Gori, B C Beard, C Ironside, G Karponi & H-P Kiem

  2. Department of Medicine, University of Washington School of Medicine, University of Washington, Seattle, WA, USA

    B C Beard & H-P Kiem

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Gori, J., Beard, B., Ironside, C. et al. In vivo selection of autologous MGMT gene-modified cells following reduced-intensity conditioning with BCNU and temozolomide in the dog model. Cancer Gene Ther 19, 523–529 (2012). https://doi.org/10.1038/cgt.2012.25

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