In Vivo Effects of Myeloablative Alkylator Therapy on Survival and Differentiation of MGMTP140K-Transduced Human G-CSF-Mobilized Peripheral Blood Cells

Shanbao Cai, Jennifer R. Hartwell, Ryan J. Cooper, Beth E. Juliar, Emi Kreklau, Rafat Abonour, W. Scott Goebel, Karen E. Pollok

Research output: Contribution to journalArticle

13 Scopus citations

Abstract

High-intensity alkylator-based chemotherapy is required to eradicate tumors expressing high levels of O6-methylguanine DNA methyltransferase (MGMT). This treatment, however, can lead to life-threatening myelosuppression. We investigated a gene therapy strategy to protect human granulocyte colony-stimulating factor-mobilized peripheral blood CD34+ cells (MPB) from a high-intensity alkylator-based regimen. We transduced MPB with an oncoretroviral vector that coexpresses MGMTP140K and the enhanced green fluorescent protein (EGFP) (n = 5 donors). At 4 weeks posttransplantation into nonobese diabetic/severe combined immunodeficient (NOD/SCID) mice, cohorts were not treated or were treated with low- or high-intensity alkylating chemotherapy. In the high-intensity-treated cohort, it was necessary to infuse NOD/SCID bone marrow (BM) to alleviate hematopoietic toxicity. At 8 weeks posttreatment, human CD45+ cells in the BM of mice treated with either regimen were EGFP+ and contained MGMT-specific DNA repair activity. In cohorts receiving low-intensity therapy, both primitive and mature hematopoietic cells were present in the BM. Although B-lymphoid and myeloid cells were resistant to in vivo drug treatment in cohorts that received high-intensity therapy, no human CD34+ cells or B-cell precursors were detected. These data suggest that improved strategies to optimize repair of DNA damage in primitive human hematopoietic cells are needed when using high-intensity anti-cancer therapy.

Original languageEnglish (US)
Pages (from-to)1016-1026
Number of pages11
JournalMolecular Therapy
Volume13
Issue number5
DOIs
StatePublished - May 1 2006

Keywords

  • BCNU
  • G-CSF-mobilized peripheral blood
  • gene therapy
  • hematopoietic stem cell
  • NOD/SCID mice
  • O-benzylguanine
  • O-methylguanine DNA methyltransferase
  • SCID-repopulating cell

ASJC Scopus subject areas

  • Molecular Biology

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