Protection and in vivo selection of hematopoietic stem cells using temozolomide, O6-benzylguanine, and an alkyltransferase-expressing retroviral vector

Nobukuni Sawai, Sheng Zhou, Elio F. Vanin, Peter Houghton, Thomas P. Brent, Brian P. Sorrentino

Research output: Contribution to journalArticlepeer-review

102 Scopus citations

Abstract

Transfer of drug resistance genes to hematopoietic stem cells offers the potential to protect cancer patients from drug-induced myelosuppression and to increase the number of gene-modified cells by in vivo selection. In this study, a retroviral vector expressing both a P140K variant of human O6-methylguanine-DNA methyltransferase (MGMT) and an EGFP reporter gene was evaluated for stem cell protection in a murine transplant model. Mice transplanted with vector-transduced cells showed significant resistance to the myelosuppressive effects of temozolomide (TMZ), an orally administered DNA-methylating drug, and O6-benzylguanine (BG), a drug that depletes cells of wild-type MGMT activity. Following drug treatment, increases in EGFP+ peripheral blood cells were seen in all peripheral blood lineages, and secondary transplant experiments proved that selection had occurred at the stem cell level. In a second set of experiments in which transduced cells were diluted with unmarked cells, efficient stem cell selection was noted together with progressive marrow protection with repeated treatment courses. Altogether, these results show that P140K MGMT gene transfer can protect stem cells against the toxic effects of TMZ and BG and that this vector/drug system may be useful for clinical myeloprotection and for in vivo selection of transduced stem cells.

Original languageEnglish (US)
Pages (from-to)78-87
Number of pages10
JournalMolecular Therapy
Volume3
Issue number1
DOIs
StatePublished - 2001

Keywords

  • Bone marrow transplantation
  • Gene therapy
  • Hematopoietic stem cells
  • Methylguanine-DNA methyltransferase
  • O-benzylguanine
  • Retroviral vector
  • Temozolomide

ASJC Scopus subject areas

  • Molecular Medicine
  • Molecular Biology
  • Genetics
  • Pharmacology
  • Drug Discovery

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