title: GST-pi gene-transduced hematopoietic progenitor cell transplantation overcomes the bone marrow toxicity of cyclophosphamide in mice.
abstract: Autologous transplantation of bone marrow cells (BMCs) transduced with the multidrug resistance 1 (MDR1) gene or dihydrofolate reductase (DHFR) gene has already been applied in clinical chemoprotection trials. However, anticancer drugs frequently used in high-dose chemotherapy (HDC), such as alkylating agents, are not relevant to MDR1 or DHFR gene products. In this context, we have previously reported that glutathione S-transferase-pi (GST-pi) gene-transduced human CD34(+) cells showed resistance in vitro against 4-hydroperoxicyclophosphamide, an active form of cyclophosphamide (CY). In the present study, a subsequent attempt was made in a murine model to evaluate the effectiveness of transplantation of GST-pi-transduced BMCs to protect bone marrow against high-dose CY. The gene transfection was carried out retrovirally, employing a recombinant fibronectin fragment. Transfection efficiency into CFU-GM was 30%. After the transplantation, recipient mice (GST-pi mice) received three sequential courses of high-dose CY. As the chemotherapy courses advanced, both shortening of recovery period from WBC nadir and shallowing of WBC nadir were observed. In contrast to the fact that three of seven control mice died, possibly due to chemotoxicity, all seven GST-pi mice were alive after the third course, at which point the vector GST-pi gene was detected in 50% of CFU-GM derived from their BMCs and peripheral blood mononuclear cells. When BMCs obtained from these seven mice were retransplanted into secondary recipient mice, 20% of CFU-GM from BMCs showed positive signals for vector GST-pi DNA after 6 months. These data indicate that the GST-pi gene can confer resistance to bone marrow against CY by being transduced into long-term repopulating cells.
authors.affiliation: Fourth Department of Internal Medicine, Sapporo Medical University School of Medicine, Japan.
authors.forename: T
authors.initials: T
authors.lastname: Matsunaga
authors.forename: S
authors.initials: S
authors.lastname: Sakamaki
authors.forename: T
authors.initials: T
authors.lastname: Kuga
authors.forename: H
authors.initials: H
authors.lastname: Kuroda
authors.forename: T
authors.initials: T
authors.lastname: Kusakabe
authors.forename: T
authors.initials: T
authors.lastname: Akiyama
authors.forename: Y
authors.initials: Y
authors.lastname: Konuma
authors.forename: Y
authors.initials: Y
authors.lastname: Hirayama
authors.forename: M
authors.initials: M
authors.lastname: Kobune
authors.forename: J
authors.initials: J
authors.lastname: Kato
authors.forename: K
authors.initials: K
authors.lastname: Sasaki
authors.forename: K
authors.initials: K
authors.lastname: Kogawa
authors.forename: R
authors.initials: R
authors.lastname: Koyama
authors.forename: Y
authors.initials: Y
authors.lastname: Niitsu
mesh_terms: D000818:Animals; D018906:Antineoplastic Agents, Alkylating / Q000633:toxicity*; D001853:Bone Marrow / Q000187:drug effects*; D003520:Cyclophosphamide / Q000633:toxicity*; D005260:Female; D018014:Gene Transfer Techniques; D051549:Glutathione S-Transferase pi; D005982:Glutathione Transferase / Q000235:genetics* / Q000378:metabolism; D018380:Hematopoietic Stem Cell Transplantation; D006412:Hematopoietic Stem Cells / Q000187:drug effects*; D006801:Humans; D007527:Isoenzymes / Q000235:genetics* / Q000378:metabolism; D007958:Leukocyte Count; D051379:Mice; D008807:Mice, Inbred BALB C; D016133:Polymerase Chain Reaction
chemical_list: D018906:Antineoplastic Agents, Alkylating; D007527:Isoenzymes; D003520:Cyclophosphamide; C496556:GSTP1 protein, human; D051549:Glutathione S-Transferase pi; D005982:Glutathione Transferase; C496557:Gstp1 protein, mouse