In vivo expansion of MDR1-transduced cells accompanied by a post-transplantation chemotherapy regimen with mitomycin C and methotrexate

Junko Mitsuhashi, Hiroyo Hosoyama, Satomi Tsukahara, Kazuhiro Katayama, Kohji Noguchi, Yoshinori Ito, Kiyohiko Hatake, Keisuke Aiba, Shunji Takahashi, Yoshikazu Sugimoto

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

Background: A recurrent breast cancer patient received high-dose chemotherapy, a transplant of multidrug resistance 1 (MDR1)-transduced cells and four different protocols of post-transplantation chemotherapy. We report the analysis of MDR1-transduced cells in this patient. Methods: MDR1 transgene levels in the peripheral blood mononuclear cells of the patient were evaluated by polymerase chain reaction (PCR). Retroviral integration sites of the MDR1-transduced cells were identified by linear amplification-mediated (LAM)-PCR. Results: Twelve days after transplantation, approximately 1% of the peripheral blood mononuclear cells were MDR1 transgene-positive. The transgene levels decreased quickly, and were at low levels until day 504. A remarkable increase in MDR1 transgene-positive cells was observed on day 532, during combination chemotherapy with mitomycin C and methotrexate. Using LAM-PCR, 31 MDR1-transduced clones were identified, and eight of these were long-life clones that survived for more than 500 days. Among the 31 clones, ten had a retroviral integration site near genes listed in the Retroviral Tagged Cancer Gene (RTCG) Database. Two long-life clones, N-30 and N-31, had retroviral integration sites within the MDS1-EVI1 locus. Another two long-life clones had integration sites close to PRDM16 or CUEDC1. Conclusions: These results suggest that MDR1-transduced cells were enriched in vivo by an MDR1 substrate, mitomycin C. The possible activation of EVI1 or other RTCGs by retroviral insertion may have affected the survival and persistence of a proportion of the transduced cells.

Original languageEnglish
Pages (from-to)596-603
Number of pages8
JournalJournal of Gene Medicine
Volume12
Issue number7
DOIs
Publication statusPublished - 2010 Jul

Fingerprint

Multiple Drug Resistance
Mitomycin
Methotrexate
Transplantation
Drug Therapy
Transgenes
Clone Cells
Polymerase Chain Reaction
Blood Cells
Neoplasm Genes
Combination Drug Therapy
Databases
Breast Neoplasms
Transplants
Survival

Keywords

  • Breast cancer
  • CD34
  • Gene therapy
  • Hematopoietic stem cell
  • P-glycoprotein
  • Retrovirus

ASJC Scopus subject areas

  • Genetics
  • Molecular Biology
  • Molecular Medicine
  • Genetics(clinical)
  • Drug Discovery
  • Medicine(all)

Cite this

In vivo expansion of MDR1-transduced cells accompanied by a post-transplantation chemotherapy regimen with mitomycin C and methotrexate. / Mitsuhashi, Junko; Hosoyama, Hiroyo; Tsukahara, Satomi; Katayama, Kazuhiro; Noguchi, Kohji; Ito, Yoshinori; Hatake, Kiyohiko; Aiba, Keisuke; Takahashi, Shunji; Sugimoto, Yoshikazu.

In: Journal of Gene Medicine, Vol. 12, No. 7, 07.2010, p. 596-603.

Research output: Contribution to journalArticle

Mitsuhashi, J, Hosoyama, H, Tsukahara, S, Katayama, K, Noguchi, K, Ito, Y, Hatake, K, Aiba, K, Takahashi, S & Sugimoto, Y 2010, 'In vivo expansion of MDR1-transduced cells accompanied by a post-transplantation chemotherapy regimen with mitomycin C and methotrexate', Journal of Gene Medicine, vol. 12, no. 7, pp. 596-603. https://doi.org/10.1002/jgm.1474
Mitsuhashi, Junko ; Hosoyama, Hiroyo ; Tsukahara, Satomi ; Katayama, Kazuhiro ; Noguchi, Kohji ; Ito, Yoshinori ; Hatake, Kiyohiko ; Aiba, Keisuke ; Takahashi, Shunji ; Sugimoto, Yoshikazu. / In vivo expansion of MDR1-transduced cells accompanied by a post-transplantation chemotherapy regimen with mitomycin C and methotrexate. In: Journal of Gene Medicine. 2010 ; Vol. 12, No. 7. pp. 596-603.
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abstract = "Background: A recurrent breast cancer patient received high-dose chemotherapy, a transplant of multidrug resistance 1 (MDR1)-transduced cells and four different protocols of post-transplantation chemotherapy. We report the analysis of MDR1-transduced cells in this patient. Methods: MDR1 transgene levels in the peripheral blood mononuclear cells of the patient were evaluated by polymerase chain reaction (PCR). Retroviral integration sites of the MDR1-transduced cells were identified by linear amplification-mediated (LAM)-PCR. Results: Twelve days after transplantation, approximately 1{\%} of the peripheral blood mononuclear cells were MDR1 transgene-positive. The transgene levels decreased quickly, and were at low levels until day 504. A remarkable increase in MDR1 transgene-positive cells was observed on day 532, during combination chemotherapy with mitomycin C and methotrexate. Using LAM-PCR, 31 MDR1-transduced clones were identified, and eight of these were long-life clones that survived for more than 500 days. Among the 31 clones, ten had a retroviral integration site near genes listed in the Retroviral Tagged Cancer Gene (RTCG) Database. Two long-life clones, N-30 and N-31, had retroviral integration sites within the MDS1-EVI1 locus. Another two long-life clones had integration sites close to PRDM16 or CUEDC1. Conclusions: These results suggest that MDR1-transduced cells were enriched in vivo by an MDR1 substrate, mitomycin C. The possible activation of EVI1 or other RTCGs by retroviral insertion may have affected the survival and persistence of a proportion of the transduced cells.",
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AU - Katayama, Kazuhiro

AU - Noguchi, Kohji

AU - Ito, Yoshinori

AU - Hatake, Kiyohiko

AU - Aiba, Keisuke

AU - Takahashi, Shunji

AU - Sugimoto, Yoshikazu

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AB - Background: A recurrent breast cancer patient received high-dose chemotherapy, a transplant of multidrug resistance 1 (MDR1)-transduced cells and four different protocols of post-transplantation chemotherapy. We report the analysis of MDR1-transduced cells in this patient. Methods: MDR1 transgene levels in the peripheral blood mononuclear cells of the patient were evaluated by polymerase chain reaction (PCR). Retroviral integration sites of the MDR1-transduced cells were identified by linear amplification-mediated (LAM)-PCR. Results: Twelve days after transplantation, approximately 1% of the peripheral blood mononuclear cells were MDR1 transgene-positive. The transgene levels decreased quickly, and were at low levels until day 504. A remarkable increase in MDR1 transgene-positive cells was observed on day 532, during combination chemotherapy with mitomycin C and methotrexate. Using LAM-PCR, 31 MDR1-transduced clones were identified, and eight of these were long-life clones that survived for more than 500 days. Among the 31 clones, ten had a retroviral integration site near genes listed in the Retroviral Tagged Cancer Gene (RTCG) Database. Two long-life clones, N-30 and N-31, had retroviral integration sites within the MDS1-EVI1 locus. Another two long-life clones had integration sites close to PRDM16 or CUEDC1. Conclusions: These results suggest that MDR1-transduced cells were enriched in vivo by an MDR1 substrate, mitomycin C. The possible activation of EVI1 or other RTCGs by retroviral insertion may have affected the survival and persistence of a proportion of the transduced cells.

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