Retrovirus-mediated gene transfer of rat glutathione S-transferase Yc confers in vitro resistance to alkylating agents in human leukemia cells and in clonogenic mouse hematopoietic progenitor cells.
Recently, we have reported that N2Yc, a Moloney-based retrovirus vector expressing the Yc isoform of rat glutathione S-transferase (GST-Yc), conferred resistance to alkylating agents in mouse NIH-3T3 fibroblasts. In this report, we address the feasibility of using rat GST-Yc somatic gene transfer to confer chemoprotection to the hematopoietic system. Human chronic myelogenous leukemia K-562 cells were efficiently transduced with the N2Yc retrovirus vector and showed a significant increase in the 50% inhibitory concentration of chlorambucil (3.2- to 3.3-fold), mechlorethamine (4.7- to 5.3-fold), and melphalan (2.1- to 2.2-fold). In addition, primary murine clonogenic hematopoietic progenitor cells transduced with the N2Yc vector were significantly more resistant to alkylating agents in vitro than cells transduced with the antisense N2revYc vector. The survival of Yc-transduced hematopoietic colonies at 400 nM mechlorethamine and 4 mu M chlorambucil was 39.4% and 42.6%, respectively, compared to 27.2% and 30.4% for N2revYc-transduced cells. Future experiments will determine the level of chemoprotection achievable in vivo, following transplantation of N2Yc-transduced hematopoietic cells in mice.
Letourneau
S
S
Department of Medicine, Montreal General Hospital, Montreal, Canada.
Greenbaum
M
M
Cournoyer
D
D
D000477:Alkylating Agents / Q000494:pharmacology*; D000818:Animals; D015139:Blotting, Southern; D015153:Blotting, Western; D016026:Bone Marrow Transplantation; D002699:Chlorambucil / Q000494:pharmacology; D002999:Clone Cells; D019008:Drug Resistance, Neoplasm / Q000235:genetics*; D005260:Female; D018014:Gene Transfer Techniques; D005978:Glutathione / Q000378:metabolism; D005979:Glutathione Peroxidase / Q000378:metabolism; D005982:Glutathione Transferase / Q000187:drug effects / Q000235:genetics* / Q000378:metabolism; D006412:Hematopoietic Stem Cells / Q000187:drug effects / Q000201:enzymology / Q000821:virology; D006801:Humans; D007938:Leukemia / Q000188:drug therapy / Q000235:genetics / Q000473:pathology; D008466:Mechlorethamine / Q000494:pharmacology; D051379:Mice; D008809:Mice, Inbred C3H; D051381:Rats; D012190:Retroviridae / Q000235:genetics*; D014158:Transcription, Genetic; D014161:Transduction, Genetic; D014407:Tumor Cells, Cultured
D000477:Alkylating Agents; D002699:Chlorambucil; D008466:Mechlorethamine; D005979:Glutathione Peroxidase; D005982:Glutathione Transferase; D005978:Glutathione