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Vol. 110, No. 2-3, 2003
Issue release date: October 2003
Acta Haematol 2003;110:93–106

Hematoprotection by Transfer of Drug-Resistance Genes

Flasshove M. · Moritz T. · Bardenheuer W. · Seeber S.
Department of Internal Medicine (Cancer Research), West German Cancer Center, University of Essen Medical School, Essen, Germany

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Myelosuppression represents a major side effect of cytotoxic anti-cancer agents. Infection due to granulocytopenia and the risk of bleeding due to thrombocytopenia compromise the potential of curative and palliative chemotherapy. Considering the many chemotherapeutic agents for which drug resistance genes have been described, and the recent improvements in vector and transduction technology, it seems conceivable that drug resistance gene transfer into a patient’s autologous hematopoietic stem or progenitor cells will be able to reduce or abolish chemotherapy-induced myelosuppression.

Copyright / Drug Dosage

Copyright: All rights reserved. No part of this publication may be translated into other languages, reproduced or utilized in any form or by any means, electronic or mechanical, including photocopying, recording, microcopying, or by any information storage and retrieval system, without permission in writing from the publisher or, in the case of photocopying, direct payment of a specified fee to the Copyright Clearance Center.
Drug Dosage: The authors and the publisher have exerted every effort to ensure that drug selection and dosage set forth in this text are in accord with current recommendations and practice at the time of publication. However, in view of ongoing research, changes in goverment regulations, and the constant flow of information relating to drug therapy and drug reactions, the reader is urged to check the package insert for each drug for any changes in indications and dosage and for added warnings and precautions. This is particularly important when the recommended agent is a new and/or infrequently employed drug.
Disclaimer: The statements, opinions and data contained in this publication are solely those of the individual authors and contributors and not of the publishers and the editor(s). The appearance of advertisements or/and product references in the publication is not a warranty, endorsement, or approval of the products or services advertised or of their effectiveness, quality or safety. The publisher and the editor(s) disclaim responsibility for any injury to persons or property resulting from any ideas, methods, instructions or products referred to in the content or advertisements.


  1. Attal M, Harousseau JL, Stoppa AM, Sotto JJ, Fuzibet JG, Rossi JF, Casassus P, Maisonneuve H, Facon T, Ifrah N, Payen C, Bataille R: A prospective, randomized trial of autologous bone marrow transplantation and chemotherapy in multiple myeloma. N Engl J Med 1996;335:91–97.
  2. Philip T, Guglielmi C, Hagenbeek A, Somers R, Van der Lelie H, Bron D, Sonneveld P, Gisselbrecht C, Cahn JY, Harousseau JL, Coiffier B, Biron P, Mandelli F, Chauvin F: Autologous bone marrow transplantation as compared with salvage chemotherapy in relapses of chemotherapy-sensitive non-Hodgkin’s lymphoma. N Engl J Med 1995;333:1540–1545.
  3. Sobecks RM, Vogelzang NJ: High-dose chemotherapy with autologous stem-cell support for germ cell tumors: A critical review. Semin Oncol 1999;26:106–118.
  4. Bokemeyer C, Kollmannsberger C, Meisner C, Harstrick A, Beyer J, Metzner B, Hartmann JT, Schmoll HJ, Einhorn L, Kanz L, Nichols C: First-line high-dose chemotherapy compared with standard-dose PEB/VIP chemotherapy in patients with advanced germ cell tumors: A multivariate and matched-pair analysis. J Clin Oncol 1999;17:3450–3456.
  5. Rodenhuis S: High-dose chemotherapy in breast cancer – Interpretation of the randomized trials. Anticancer Drugs 2001;12:85–88.
  6. Cavazzana-Calvo M, Hacein-Bey S, de Saint Basile G, Gross F, Yvon E, Nusbaum P, Selz F, Hue C, Certain S, Casanova JL, Bousso P, Le Deist F, Fischer A: Gene therapy of human severe combined immunodeficiency (SCID)-X1 disease. Science 2000;288:669–672.
  7. Joyner A, Keller G, Phillips RA, Bernstein A: Retrovirus transfer of a bacterial gene into mouse haematopoietic progenitor cells. Nature 1983;305:556–558.
  8. Hock RA, Miller AD: Retrovirus-mediated transfer and expression of drug resistance genes in human haematopoietic progenitor cells. Nature 1986;320:275–277.
  9. Belmont JW, Henkel-Tigges J, Chang SMW, Wager-Smith K, Kellems RE, Dick JE, Magli MC, Phillips RA, Bernstein A, Caskey CT: Expression of human adenosine deaminase in murine haematopoietic progenitor cells following retroviral transfer. Nature 1986;322:385–387.
  10. Keller G, Paige C, Gilboa E, Wagner EF: Expression of a foreign gene in myeloid and lymphoid cells derived from multipotent haematopoietic precursors. Nature 1985;318:149–154.
  11. Miller AD, Eckner RJ, Jolly DJ, Friedmann T, Verma IM: Expression of a retrovirus encoding human HPRT in mice. Science 1984;225:630–632.
  12. Williams DA, Lemischka IR, Nathan DG, Mulligan RC: Introduction of new genetic material into pluripotent haematopietic stem cells of the mouse. Nature 1984;310:476–480.
  13. Halene S, Kohn DB: Gene therapy using hematopoietic stem cells: Sisyphus approaches the crest. Hum Gene Ther 2000;11:1259–1267.

    External Resources

  14. Gilboa E, Eglitis MA, Kantoff PW, Anderson WF: Transfer and expression of cloned genes using retroviral vectors. Biotechniques 1986;4:504–512.
  15. Miller AD, Rosman GJ: Improved retroviral vectors for gene transfer and expression. Biotechniques 1989;7:980–990.
  16. Baum C, Hegewisch-Becker S, Eckert HG, Stocking C, Ostertag W: Novel retroviral vectors for efficient expression of the multidrug resistance (mdr-1) gene in early hematopoietic cells. J Virol 1995;69:7541–7547.
  17. Buchschacher GL, Wong-Staal F: Development of lentiviral vectors for gene therapy for human diseases. Blood 2000;95:2499–2504.
  18. May C, Rivella S, Callegari J, Heller G, Gaensler KM, Luzzatto L, Sadelain M: Therapeutic haemoglobin synthesis in beta-thalassaemic mice expressing lentivirus-encoded human beta-globin. Nature 2000;406:82–86.
  19. Chu E, Mota AC, Fogarasi MC: Pharmacology of cancer chemotherapy: Antimetabolites; in DeVita VT Jr, Hellman S, Rosenberg SA (eds): Cancer, ed 6. Philadelphia, Lippincott Williams & Wilkins, 2001, pp 388–414.
  20. Simonsen CC, Levinson AD: Isolation and expression of an altered mouse dihydrofolate reductase cDNA. Proc Natl Acad Sci USA 1983;80:2495–2499.
  21. Thillet J, Absil J, Stone SR, Pictet R: Site-directed mutagenesis of mouse dihydrofolate reductase. Mutants with increased resistance to methotrexate and trimethoprim. J Biol Chem 1988;263:12500–12508.
  22. Schweitzer BI, Srimatkandada S, Gritsman H, Sheridan R, Venkataraghavan R, Bertino JR: Probing the role of two hydrophobic active site residues in the human dihydrofolate reductase by site-directed mutagenesis. J Biol Chem 1989;264:20786–20795.
  23. Ercikan-Abali EA, Mineishi S, Tong Y, Nakahara S, Waltham MC, Banerjee D, Chen W, Sadelain M, Bertino JR: Active site-directed double mutants of dihydrofolate reductase. Cancer Res 1996;56:4142–4145.
  24. Patel M, Sleep SE, Lewis WS, Spencer HT, Mareya SM, Sorrentino BP, Blakley RL: Comparison of the protection of cells from antifolates by transduced human dihydrofolate reductase mutants. Hum Gene Ther 1997;8:2069–2077.
  25. Williams DA, Hsieh K, DeSilva A, Mulligan RC: Protection of bone marrow transplant recipients from lethal doses of methotrexate by the generation of methotrexate-resistant bone marrow. J Exp Med 1987;166:210–218.
  26. Kwok WW, Schuening F, Stead RB, Miller AD: Retroviral transfer of genes into canine hemopoietic progenitor cells in culture: A model for human gene therapy. Proc Natl Acad Sci USA 1986;83:4552–4555.
  27. Li M, Hantzopoulos PA, Banerjee D, Zhao SC, Schweitzer BI, Gilboa E, Bertino JR: Comparison of the expression of a mutant dihydrofolate reductase under control of different internal promoters in retroviral vectors. Hum Gene Ther 1992;3:381–390.
  28. Corey CA, DeSilva AD, Holland CA, Williams DA: Serial transplantation of methotrexate-resistant bone marrow: Protection of murine recipients from drug toxicity by progeny of transduced stem cells. Blood 1990;75:337–343.
  29. Li MX, Banerjee D, Zhao SC, Schweitzer BI, Mineishi S, Gilboa E, Bertino JR: Development of a retroviral construct containing a human mutated dihydrofolate reductase cDNA for hematopoietic stem cell transduction. Blood 1994;83:3403–3408.
  30. Zhao SC, Banerjee D, Mineishi S, Bertino JR: Post-transplant methotrexate administration leads to improved curability of mice bearing a mammary tumor transplanted with marrow transduced with a mutant human dihydrofolate reductase cDNA. Hum Gene Ther 1997;8:903–909.
  31. Flasshove M, Banerjee D, Leonard JP, Mineishi S, Li MX, Bertino JR, Moore MAS: Retroviral transduction of human CD34+ umbilical cord blood progenitor cells with a mutated dihydrofolate reductase cDNA. Hum Gene Ther 1998;9:63–71.
  32. Flasshove M, Banerjee D, Mineishi S, Li MX, Bertino JR, Moore MAS: Ex vivo expansion and selection of human CD34+ peripheral blood progenitor cells after introduction of a mutated dihydrofolate reductase cDNA via retroviral gene transfer. Blood 1995;85:566–574.
  33. Beauséjour CM, Le NLO, Létourneau S, Cournoyer D, Momparler RL: Coexpression of cytidine deaminase and mutant dihydrofolate reductase by a bicistronic retroviral vector confers resistance to cytosine arabinoside and methotrexate. Hum Gene Ther 1998;9:2537–2544.
  34. Galipeau J, Benaim E, Spencer HT, Blakley RL, Sorrentino BP: A bicistronic retroviral vector for protecting hematopoietic cells against antifolates and P-glycoprotein effluxed drugs. Hum Gene Ther 1997;8:1773–1783.
  35. Sauerbrey A, McPherson JP, Zhao SC, Banerjee D, Bertino JR: Expression of a novel double-mutant dihydrofolate reductase-cytidine deaminase fusion gene confers resistance to both methotrexate and cytosine arabinoside. Hum Gene Ther 1999;10:2495–2504.
  36. Takebe N, Zhao SC, Adhikari D, Mineishi S, Sadelain M, Hilton J, Colvin M, Banerjee D, Bertino JR: Generation of dual resistance to 4-hydroperoxycyclophosphamide and methotrexate by retroviral transfer of the human aldehyde dehydrogenase class 1 gene and a mutated dihydrofolate reductase gene. Mol Ther 2001;3:88–96.
  37. Gondo H, Harada M, Taniguchi S, Akashi K, Hayashi S, Teshima T, Takamatsu Y, Eto T, Nagafuji K, Yamasaki K, Shibuya T, Niho Y: Cyclosporine combined with methylprednisolone or methotrexate in prophylaxis of moderate to severe acute graft-versus-host disease. Bone Marrow Transplant 1993;12:437–441.
  38. Sorrentino BP, Allay JA, Blakley RL: In vivo selection of hematopoietic stem cells transduced with DHFR- expressing retroviral vectors. Prog Exp Tumor Res 1999;36:143–161.
  39. Allay JA, Spencer HT, Wilkinson SL, Belt JA, Blakley RL, Sorrentino BP: Sensitization of hematopoietic stem and progenitor cells to trimetrexate using nucleoside transport inhibitors. Blood 1997;90:3546–3554.
  40. Spencer HT, Sleep SE, Rehg JE, Blakley RL, Sorrentino BP: A gene transfer strategy for making bone marrow cells resistant to trimetrexate. Blood 1996;87:2579–2587.
  41. Zhao RCH, McIvor RS, Griffin JD, Verfaillie CM: Gene therapy for chronic myelogenous leukemia (CML): A retroviral vector that renders hematopoietic progenitors methotrexate-resistant and CML progenitors functionally normal and nontumorigenic in vivo. Blood 1997;90:4687–4698.
  42. Meisel R, Bardenheuer W, Strehblow C, Seeber S, Moritz T, Flasshove M: Efficient protection against methotrexate (MTX) and selection of human hematopoietic cells after transfer of dihydrofolate reductase (DHFR) mutants (abstract). Blood 2001;98:405b.
  43. Gatlin J, Douglas J, Evans JT, Collins RH, Wendel GD, Garcia JV: In vitro selection of lentivirus vector-transduced human CD34+ cells. Hum Gene Ther 2000;11:1949–1957.

    External Resources

  44. Ensminger WD, Frei E: The prevention of methotrexate toxicity by thymidine infusions in humans. Cancer Res 1977;37:1857–1863.
  45. Pinedo HM, Zaharko DS, Bull JM, Chabner BA: The reversal of methotrexate cytotoxicity to mouse bone marrow cells by leucovorin and nucleosides. Cancer Res 1976;36:4418–4424.
  46. Allay JA, Persons DA, Galipeau J, Riberdy JM, Ashmun RA, Blakley RL, Sorrentino BP: In vivo selection of retrovirally transduced hematopoietic stem cells. Nat Med 1998;4:1136–1143.
  47. Blau CA, Neff T, Papayannopoulou T: The hematological effects of folate analogs: Implications for using the dihydrofolate reductase gene for in vivo selection. Hum Gene Ther 1996;7:2069–2078.
  48. May C, Gunther R, McIvor RS: Protection of mice from lethal doses of methotrexate by transplantation with transgenic marrow expressing drug-resistant dihydrofolate reductase activity. Blood 1995;86:2439–2448.
  49. Takebe N, Nakahara S, Zhao SC, Adhikari D, Ural AU, Iwamoto M, Banerjee D, Bertino JR: Comparison of methotrexate resistance conferred by a mutated dihydrofolate reductase (DHFR) cDNA in two different retroviral vectors. Cancer Gene Ther 2000;7:910–919.
  50. Takebe N, Xu LC, MacKenzie KL, Bertino JR, Moore MA: Methotrexate selection of long-term culture initiating cells following transduction of CD34(+) cells with a retrovirus containing a mutated human dihydrofolate reductase gene. Cancer Gene Ther 2002;9:308–320.
  51. Mineishi S, Nakahara S, Takebe N, Banerjee D, Zhao SC, Bertino JR: Co-expression of the herpes simplex virus thymidine kinase gene potentiates methotrexate resistance conferred by transfer of a mutated dihydrofolate reductase gene. Gene Ther 1997;4:570–576.
  52. Mineishi S, Nakahara S, Takebe N, Zhao SC, Banerjee D, Bertino JR: Purine salvage rescue by xanthine-guanine phosphoribosyltransferase (XGPRT) potentiates methotrexate resistance conferred by transfer of a mutated dihydrofolate reductase gene. Cancer Gene Ther 1998;5:144–149.
  53. Patel DH, Allay JA, Belt JA, Sorrentino BP: Retroviral transfer of the hENT2 nucleoside transporter cDNA confers broad-spectrum antifolate resistance in murine bone marrow cells. Blood 2000;95:2356–2363.
  54. Tong Y, Liu-Chen X, Ercikan-Abali EA, Zhao SC, Banerjee D, Maley F, Bertino JR: Probing the folate-binding site of human thymidylate synthase by site-directed mutagenesis. Generation of mutants that confer resistance to raltitrexed, Thymitaq, and BW1843U89. J Biol Chem 1998;273:31209–31214.
  55. Landis DM, Heindel CC, Loeb LA: Creation and characterization of 5-fluorodeoxyuridine-resistant Arg50 loop mutants of human thymidylate synthase. Cancer Res 2001;61:666–672.
  56. Shaw D, Berger FG, Spencer HT: Retroviral expression of Escherichia coli thymidylate synthase cDNA confers high-level antifolate resistance to hematopoietic cells. Hum Gene Ther 2001;12:51–59.
  57. Pastan I, Gottesman MM, Ueda K, Lovelace E, Rutherford AV, Willingham MC: A retrovirus carrying an MDR1 cDNA confers multidrug resistance and polarized expression of P-glycoprotein in MDCK cells. Proc Natl Acad Sci USA 1988;85:4486–4490.
  58. Sorrentino BP, Brandt SJ, Bodine D, Gottesman M, Pastan I, Cline A, Nienhuis AW: Selection of drug-resistant bone marrow cells in vivo after retroviral transfer of human MDR1. Science 1992;257:99–103.
  59. Hanania EG, Fu S, Zu Z, Hegewisch-Becker S, Korbling M, Hester J, Durett A, Andreeff M, Mechetner E, Holzmayer T, et al.: Chemotherapy resistance to taxol in clonogenic progenitor cells following transduction of CD34 selected marrow and peripheral blood cells with a retrovirus that contains the MDR-1 chemotherapy resistance gene. Gene Ther 1995;2:285–294.
  60. Eckert HG, Stockschläder M, Just U, Hegewisch-Becker S, Grez M, Uhde A, Zander A, Ostertag W, Baum C: High-dose multidrug resistance in primary human hematopoietic progenitor cells transduced with optimized retroviral vectors. Blood 1996;88:3407–3415.
  61. Bodine DM, Seidel NE, Gale MS, Nienhuis AW, Orlic D: Efficient retrovirus transduction of mouse pluripotent hematopoietic stem cells mobilized into the peripheral blood by treatment with granulocyte colony-stimulating factor and stem cell factor. Blood 1994;84:1482–1491.
  62. Hildinger M, Fehse B, Hegewisch-Becker S, John J, Rafferty JR, Ostertag W, Baum C: Dominant selection of hematopoietic progenitor cells with retroviral MDR1 co-expression vectors. Hum Gene Ther 1998;9:33–42.
  63. Bertolini F, DeMonte L, Corsini C, Lazzari L, Lauri L, Soligo D, Ward M, Bank A, Malavasi F: Retrovirus-mediated transfer of the multidrug resistance gene into human haemopoietic progenitor cells. Br J Haematol 1994;88:318–324.
  64. Ward M, Richardson C, Pioli P, Smith L, Podda S, Goff S, Hesdorffer C, Bank A: Transfer and expression of the human multiple drug resistance gene in human CD34+ cells. Blood 1994;84:1408–1414.
  65. Licht T, Aksentijevich I, Gottesman MM, Pastan I: Efficient expression of functional human MDR1 gene in murine bone marrow after retroviral transduction of purified hematopoietic stem cells. Blood 1995;86:111–121.
  66. Sorrentino BP, McDonagh KT, Woods D, Orlic D: Expression of retroviral vectors containing the human multidrug resistance 1 cDNA in hematopoietic cells of transplanted mice. Blood 1995;86:491–501.
  67. Licht T, Aran JM, Goldenberg SK, Vieira WD, Gottesman MM, Pastan I: Retroviral transfer of human MDR1 gene to hematopoietic cells: Effects of drug selection and of transcript splicing on expression of encoded P-glycoprotein. Hum Gene Ther 1999;10:2173–2185.
  68. Knipper R, Kuehlcke K, Schiedlmeier B, Hildinger M, Lindemann C, Schilz AJ, Fauser AA, Fruehauf S, Zeller WJ, Ostertag W, Eckert HG, Baum C: Improved post-transcriptional processing of an MDR1 retrovirus elevates expression of multidrug resistance in primary human hematopoietic cells. Gene Ther 2001;8:239–246.
  69. Schiedlmeier B, Kühlcke K, Eckert HG, Baum C, Zeller WJ, Fruehauf S: Quantitative assessment of retroviral transfer of the human multidrug resistance 1 gene to human mobilized peripheral blood progenitor cells engrafted in nonobese diabetic/severe combined immunodeficient mice. Blood 2000;95:1237–1248.
  70. Licht T, Goldenberg SK, Vieira WD, Gottesman MM, Pastan I: Drug selection of MDR1-transduced hematopoietic cells ex vivo increases transgene expression and chemoresistance in reconstituted bone marrow in mice. Gene Ther 2000;7:348–358.
  71. Bertolini F, Battaglia M, Lanza A, Gibelli N, Palermo B, Pavesi L, Caprotti M, Robustelli della Cuna G: Multilineage long-term engraftment potential of drug-resistant hematopoietic progenitors. Blood 1997;90:3027–3036.
  72. Chaudhary PM, Roninson IB: Expression and activity of P-glycoprotein, a multidrug efflux pump, in human hematopoietic stem cells. Cell 1991;66:85–94.
  73. Cowan KH, Moscow JA, Huang H, Zujewski JA, O’Shaughnessy J, Sorrentino B, Hines K, Carter C, Schneider E, Cusack G, Noone M, Dunbar C, Steinberg S, Wilson W, Goldspiel B, Read EJ, Leitman SF, McDonagh K, Chow C, Abati A, Chiang Y, Chang YN, Gottesman MM, Pastan I, Nienhuis A: Paclitaxel chemotherapy after autologous stem-cell transplantation and engraftment of hematopoietic cells transduced with a retrovirus containing the multidrug resistance complementary DNA (MDR1) in metastatic breast cancer patients. Clin Cancer Res 1999;5:1619–1628.
  74. Devereux S, Corney C, Macdonald C, Watts M, Sullivan A, Goldstone AH, Ward M, Bank A, Linch DC: Feasibility of multidrug resistance (MDR-1) gene transfer in patients undergoing high-dose therapy and peripheral blood stem cell transplantation for lymphoma. Gene Ther 1998;5:403–408.
  75. Hanania EG, Giles RE, Kavanagh J, Ellerson D, Zu Z, Wang T, Su Y, Kudelka A, Rahman Z, Holmes F, Hortobagyi G, Claxton D, Bachier C, Thall P, Cheng S, Hester J, Ostrove JM, Bird RE, Chang A, Korbling M, Seong D, Cote R, Holzmayer T, Deisseroth AB, et al: Results of MDR-1 vector modification trial indicate that granulocyte/macrophage colony-forming unit cells do not contribute to posttransplant hematopoietic recovery following intensive systemic therapy. Proc Natl Acad Sci USA 1996;93:15346–15351.
  76. Hesdorffer C, Ayello J, Ward M, Kaubisch A, Vahdat L, Balmaceda C, Garrett T, Fetell M, Reiss R, Bank A, Antman K: Phase I trial of retroviral-mediated transfer of the human MDR1 gene as marrow chemoprotection in patients undergoing high-dose chemotherapy and autologous stem cell transplantation. J Clin Oncol 1998;16:165–172.
  77. Moscow JA, Huang H, Carter C, Hines K, Zujewski J, Cusack G, Chow C, Venzon D, Sorrentino B, Chiang Y, Goldspiel B, Leitman S, Read EJ, Abati A, Gottesman MM, Pastan I, Sellers S, Dunbar C, Cowan KH: Engraftment of MDR1 and NeoR gene-transduced hematopoietic cells after breast cancer chemotherapy. Blood 1999;94:52–61.
  78. Rahman Z, Kavanagh J, Champlin R, Giles R, Hanania E, Fu S, Zu Z, Mehra R, Holmes F, Kudelka A, Claxton D, Verschraegen C, Gajewski J, Andreeff M, Heimfeld S, Berenson R, Ellerson D, Calvert L, Mechetner E, Holzmayer T, Dayne A, Hamer J, Bachier C, Ostrove J, Deisseroth A, et al: Chemotherapy immediately following autologous stem-cell transplantation in patients with advanced breast cancer. Clin Cancer Res 1998;4:2717–2721.
  79. Abonour R, Williams DA, Einhorn L, Hall KM, Chen J, Coffman J, Traycoff CM, Bank A, Kato I, Ward M, Williams SD, Hromas R, Robertson MJ, Smith FO, Woo D, Mills B, Srour EF, Cornetta K: Efficient retrovirus-mediated transfer of the multidrug resistance 1 gene into autologous human long-term repopulating hematopoietic stem cells. Nat Med 2000;6:652–658.
  80. Bunting KD, Galipeau J, Topham D, Benaim E, Sorrentino BP: Transduction of murine bone marrow cells with an MDR1 vector enables ex vivo stem cell expansion, but these expanded grafts cause a myeloproliferative syndrome in transplanted mice. Blood 1998;92:2269–2279.
  81. Bunting KD, Zhou S, Lu T, Sorrentino BP: Enforced P-glycoprotein pump function in murine bone marrow cells results in expansion of side population stem cells in vitro and repopulating cells in vivo. Blood 2000;96:902–909.
  82. Eckert HG, Kühlcke K, Schilz AJ, Lindemann C, Basara N, Fauser AA, Baum C: Clinical scale production of an improved retroviral vector expressing the human multidrug resistance 1 gene (MDR1). Bone Marrow Transplant 2000;26(suppl 2):S114–S117.
  83. Cole SP, Bhardwaj G, Gerlach JH, Mackie JE, Grant CE, Almquist KC, Stewart AJ, Kurz EU, Duncan AM, Deeley RG: Overexpression of a transporter gene in a multidrug-resistant human lung cancer cell line. Science 1992;258:1650–1654.
  84. Zaman GJ, Lankelma J, van Tellingen O, Beijnen J, Dekker H, Paulusma C, Oude Elferink RP, Baas F, Borst P: Role of glutathione in the export of compounds from cells by the multidrug-resistance-associated protein. Proc Natl Acad Sci USA 1995;92:7690–7694.
  85. Zaman GJ, Flens MJ, van Leusden MR, de Haas M, Mülder HS, Lankelma J, Pinedo HM, Scheper RJ, Baas F, Broxterman HJ, Borst P: The human multidrug resistance-associated protein MRP is a plasma membrane drug-efflux pump. Proc Natl Acad Sci USA 1994;91:8822–8826.
  86. D’Hondt V, Caruso M, Bank A: Retrovirus-mediated gene transfer of the multidrug resistance-associated protein (MRP) cDNA protects cells from chemotherapeutic agents. Hum Gene Ther 1997;8:1745–1751.
  87. Machiels JP, Govaerts AS, Guillaume T, Bayat B, Feyens AM, Lenoir E, Goeminne JC, Cole S, Deeley R, Caruso M, Bank A, Symann M, D’Hondt V: Retrovirus-mediated gene transfer of the human multidrug resistance-associated protein into hematopoietic cells protects mice from chemotherapy-induced leukopenia. Hum Gene Ther 1999;10:801–811.
  88. Rappa G, Lorico A, Hildinger M, Fodstad O, Baum C: Novel bicistronic retroviral vector expressing gamma-glutamylcysteine synthetase and the multidrug resistance protein 1 (MRP1) protects cells from MRP1-effluxed drugs and alkylating agents. Hum Gene Ther 2001;12:1785–1796.

    External Resources

  89. Kufe D, Spriggs D, Egan EM, Munroe D: Relationships among Ara-CTP pools, formation of (Ara-C)DNA, and cytotoxicity of human leukemic cells. Blood 1984;64:54–58.
  90. Yusa K, Ohuhara T, Tsukahara S, Tsuruo T: Human immunodeficiency virus type 1 induces 1-β-D-arabinofuranosylcytosine resistance in human H9 cell line. J Biol Chem 1992;267:16848–16850.
  91. Steuart CD, Burke PJ: Cytidine deaminase and the development of resistance to arabinosyl cytosine. Nat New Biol 1971;233:109–110.
  92. Kühn K, Bertling WM, Emmrich F: Cloning of a functional cDNA for human cytidine deaminase (CDD) and its use as a marker of monocyte/macrophage differentiation. Biochem Biophys Res Commun 1993;190:1–7.
  93. Laliberté J, Momparler RL: Human cytidine deaminase: Purification of enzyme, cloning, and expression of its complementary DNA. Cancer Res 1994;54:5401–5407.
  94. Momparler RL, Laliberté J, Eliopoulos N, Beauséjour C, Cournoyer D: Transfection of murine fibroblast cells with human cytidine deaminase cDNA confers resistance to cytosine arabinoside. Anticancer Drugs 1996;7:266–274.
  95. Schröder JK, Kirch C, Flasshove M, Kalweit H, Seidelmann M, Hilger R, Seeber S, Schütte J: Constitutive overexpression of the cytidine deaminase gene confers resistance to cytosine arabinoside in vitro. Leukemia 1996;10:1919–1924.
  96. Neff T, Blau CA: Forced expression of cytidine deaminase confers resistance to cytosine arabinoside and gemcitabine. Exp Hematol 1996;24:1340–1346.
  97. Flasshove M, Frings W, Schröder JK, Moritz T, Schütte J, Seeber S: Transfer of the cytidine deaminase cDNA into hematopoietic cells. Leuk Res 1999;23:1047–1053.
  98. Momparler RL, Eliopoulos N, Bovenzi V, Létourneau S, Greenbaum M, Cournoyer D: Resistance to cytosine arabinoside by retrovirally mediated gene transfer of human cytidine deaminase into murine fibroblast and hematopoietic cells. Cancer Gene Ther 1996;3:331–338.
  99. Eliopoulos N, Bovenzi V, Le NLO, Momparler LF, Greenbaum M, Létourneau S, Cournoyer D, Momparler RL: Retroviral transfer and long-term expression of human cytidine deaminase cDNA in hematopoietic cells following transplantation in mice. Gene Ther 1998;5:1545–1551.
  100. Beauséjour CM, Eliopoulos N, Momparler L, Le NL, Momparler RL: Selection of drug-resistant transduced cells with cytosine nucleoside analogs using the human cytidine deaminase gene. Cancer Gene Ther 2001;8:669–676.
  101. Gran C, Boyum A, Johansen RF, Lovhaug D, Seeberg EC: Growth inhibition of granulocyte-macrophage colony-forming cells by human cytidine deaminase requires the catalytic function of the protein. Blood 1998;91:4127–4135.
  102. Eliopoulos N, Cournoyer D, Momparler RL: Drug resistance to 5-aza-2′-deoxycytidine, 2′,2′-difluorodeoxycytidine, and cytosine arabinoside conferred by retroviral-mediated transfer of human cytidine deaminase cDNA into murine cells. Cancer Chemother Pharmacol 1998;42:373–378.
  103. Maze R, Carney JP, Kelley MR, Glassner BJ, Williams DA, Samson L: Increasing DNA repair methyltransferase levels via bone marrow stem cell transduction rescues mice from the toxic effects of 1,3-bis(2-chloroethyl)-1-nitrosourea, a chemotherapeutic alkylating agent. Proc Natl Acad Sci USA 1996;93:206–210.
  104. Moritz T, Mackay W, Glassner BJ, Williams DA, Samson L: Retrovirus-mediated expression of a DNA repair protein in bone marrow protects hematopoietic cells from nitrosourea-induced toxicity in vitro and in vivo. Cancer Res 1995;55:2608–2614.
  105. Allay JA, Dumenco LL, Koc ON, Liu L, Gerson SL: Retroviral transduction and expression of the human alkyltransferase cDNA provides nitrosourea resistance to hematopoietic cells. Blood 1995;85:3342–3351.
  106. Brent TP, Remack JS: Formation of covalent complexes between human O6-alkylguanine-DNA alkyltransferase and BCNU-treated defined length synthetic oligodeoxynucleotides. Nucleic Acids Res 1988;16:6779–6788.
  107. Gerson SL, Phillips W, Kastan M, Dumenco LL, Donovan C: Human CD34+ hematopoietic progenitors have low, cytokine-unresponsive O6-alkylguanine-DNA alkyltransferase and are sensitive to O6-benzylguanine plus BCNU. Blood 1996;88:1649–1655.
  108. Gerson SL, Trey JE, Miller K, Berger NA: Comparison of O6-alkylguanine-DNA alkyltransferase activity based on cellular DNA content in human, rat and mouse tissues. Carcinogenesis 1986;7:745–749.
  109. Wang G, Weiss C, Sheng P, Bresnick E: Retrovirus-mediated transfer of the human O6-methylguanine-DNA methyltransferase gene into a murine hematopoietic stem cell line and resistance to the toxic effects of certain alkylating agents. Biochem Pharmacol 1996;51:1221–1228.
  110. Jelinek J, Fairbairn LJ, Dexter TM, Rafferty JA, Stocking C, Ostertag W, Margison GP: Long-term protection of hematopoiesis against the cytotoxic effects of multiple doses of nitrosourea by retrovirus-mediated expression of human O6-alkylguanine-DNA-alkyltransferase. Blood 1996;87:1957–1961.
  111. Allay JA, Davis BM, Gerson SL: Human alkyltransferase-transduced murine myeloid progenitors are enriched in vivo by BCNU treatment of transplanted mice. Exp Hematol 1997;25:1069–1076.
  112. Dolan ME, Mitchell RB, Mummert C, Moschel RC, Pegg AE: Effect of O6-benzylguanine analogues on sensitivity of human tumor cells to the cytotoxic effects of alkylating agents. Cancer Res 1991;51:3367–3372.
  113. Spiro TP, Gerson SL, Liu L, Majka S, Haaga J, Hoppel CL, Ingalls ST, Pluda JM, Willson JK: O6-Benzylguanine: A clinical trial establishing the biochemical modulatory dose in tumor tissue for alkyltransferase-directed DNA repair. Cancer Res 1999;59:2402–2410.
  114. Reese JS, Davis BM, Liu L, Gerson SL: Simultaneous protection of G156A methylguanine DNA methyltransferase gene-transduced hematopoietic progenitors and sensitization of tumor cells using O6-benzylguanine and temozolomide. Clin Cancer Res 1999;5:163–169.
  115. Koc ON, Reese JS, Szekely EM, Gerson SL: Human long-term culture initiating cells are sensitive to benzylguanine and 1,3-bis(2-chloroethyl)-1-nitrosourea and protected after mutant (G156A) methylguanine methyltransferase gene transfer. Cancer Gene Ther 1999;6:340–348.
  116. Davis BM, Koc ON, Gerson SL: Limiting numbers of G156A O6-methylguanine-DNA methyltransferase-transduced marrow progenitors repopulate nonmyeloablated mice after drug selection. Blood 2000;95:3078–3084.
  117. Ragg S, Xu-Welliver M, Bailey J, D’Souza M, Cooper R, Chandra S, Seshadri R, Pegg AE, Williams DA: Direct reversal of DNA damage by mutant methyltransferase protein protects mice against dose-intensified chemotherapy and leads to in vivo selection of hematopoietic stem cells. Cancer Res 2000;60:5187–5195.
  118. Sawai N, Zhou S, Vanin EF, Houghton P, Brent TP, Sorrentino BP: Protection and in vivo selection of hematopoietic stem cells using temozolomide, O6-benzylguanine, and an alkyltransferase-expressing retroviral vector. Mol Ther 2001;3:78–87.
  119. Wilczynski SW, Erasmus JJ, Petros WP, Vredenburgh JJ, Folz RJ: Delayed pulmonary toxicity syndrome following high-dose chemotherapy and bone marrow transplantation for breast cancer. Am J Respir Crit Care Med 1998;157:565–573.
  120. Jansen M, Bardenheuer W, Sorg UR, Seeber S, Flasshove M, Moritz T: Protection of hematopoietic cells from O6-alkylation damage by O6-methylguanine DNA methyltransferase gene transfer: Studies with different O6-alkylating agents and retroviral backbones. Eur J Haematol 2001;67:2–13.
  121. Kobune M, Parson S, Xu Y, Kelley MR, Williams DA: Retrovirus-mediated expression of the base excision repair protein, apn1, protects hematopoietic cells from bleomycin-induced toxicity in vitro and in vivo (abstract). Blood 1999;94:381a.
  122. Kobune M, Xu Y, Baum C, Kelley MR, Williams DA: Retrovirus-mediated expression of the base excision repair proteins, formamidopyrimidine DNA glycosylase or human oxoguanine DNA glycosylase, protects hematopoietic cells from N,N′,N′′-triethylenethiophosphoramide (thioTEPA)-induced toxicity in vitro and in vivo. Cancer Res 2001;61:5116–5125.
  123. Brock N: Oxazaphosphorine cytostatics: Past-present-future. 7th Cain Memorial Award lecture. Cancer Res 1989;49:1–7.
  124. Colvin OM: Pharmacology of cancer chemotherapy: Antitumor alkylating agents; in: DeVita VT Jr, Hellman S, Rosenberg SA (eds): Cancer, ed 6. Philadelphia, Lippincott Williams & Wilkins, 2001, pp 363–376.
  125. Hilton J: Role of aldehyde dehydrogenase in cyclophosphamide-resistant L1210 leukemia. Cancer Res 1984;44:5156–5160.
  126. Kastan MB, Schlaffer E, Russo JE, Colvin OM, Civin CI, Hilton J: Direct demonstration of elevated aldehyde dehydrogenase in human hematopoietic progenitor cells. Blood 1990;75:1947–1950.
  127. Storms RW, Trujillo AP, Springer JB, Shah L, Colvin OM, Ludeman SM, Smith C: Isolation of primitive human hematopoietic progenitors on the basis of aldehyde dehydrogenase activity. Proc Natl Acad Sci USA 1999;96:9118–9123.
  128. Smith C, Gasparetto C, Collins N, Gillio A, Muench MO, O’Reilly RJ, Moore MA: Purification and partial characterization of a human hematopoietic precursor population. Blood 1991;77:2122–2128.
  129. Bunting KD, Townsend AJ: Protection by transfected rat or human class 3 aldehyde dehydrogenase against the cytotoxic effects of oxazaphosphorine alkylating agents in hamster V79 cell lines. Demonstration of aldophosphamide metabolism by the human cytosolic class 3 isozyme. J Biol Chem 1996;271:11891–11896.
  130. Bunting KD, Townsend AJ: De novo expression of transfected human class 1 aldehyde dehydrogenase (ALDH) causes resistance to oxazaphosphorine anti-cancer alkylating agents in hamster V79 cell lines. Elevated class 1 ALDH activity is closely correlated with reduction in DNA interstrand cross-linking and lethality. J Biol Chem 1996;271:11884–11890.
  131. Moreb J, Schweder M, Suresh A, Zucali JR: Overexpression of the human aldehyde dehydrogenase class I results in increased resistance to 4-hydroperoxycyclophosphamide. Cancer Gene Ther 1996;3:24–30.
  132. Moreb JS, Schweder M, Gray B, Zucali J, Zori R: In vitro selection for K562 cells with higher retrovirally mediated copy number of aldehyde dehydrogenase class-1 and higher resistance to 4-hydroperoxycyclophosphamide. Hum Gene Ther 1998;9:611–619.
  133. Magni M, Shammah S, Schiro R, Mellado W, Dalla-Favera R, Gianni AM: Induction of cyclophosphamide-resistance by aldehyde-dehydrogenase gene transfer. Blood 1996;87:1097–1103.
  134. Bunting KD, Webb M, Giorgianni F, Galipeau J, Blakley RL, Townsend AJ, Sorrentino BP: Coding region-specific destabilization of mRNA transcripts attenuates expression from retroviral vectors containing class 1 aldehyde dehydrogenase cDNAs. Hum Gene Ther 1997;8:1531–1543.
  135. Greenbaum M, Létourneau S, Assar H, Schecter RL, Batist G, Cournoyer D: Retrovirus-mediated gene transfer of rat glutathione S-transferase Yc confers alkylating drug resistance in NIH 3T3 mouse fibroblasts. Cancer Res 1994;54:4442–4447.
  136. Létourneau S, Palerme JS, Delisle JS, Beauséjour CM, Momparler RL, Cournoyer D: Coexpression of rat glutathione S-transferase A3 and human cytidine deaminase by a bicistronic retroviral vector confers in vitro resistance to nitrogen mustards and cytosine arabinoside in murine fibroblasts. Cancer Gene Ther 2000;7:757–765.
  137. Czerwinski M, Kiem HP, Slattery JT: Human CD34+ cells do not express glutathione S-transferases alpha. Gene Ther 1997;4:268–270.
  138. Kuga T, Sakamaki S, Matsunaga T, Hirayama Y, Kuroda H, Takahashi Y, Kusakabe T, Kato I, Niitsu Y: Fibronectin fragment-facilitated retroviral transfer of the glutathione-S-transferase π gene into CD34+ cells to protect them against alkylating agents. Hum Gene Ther 1997;8:1901–1910.
  139. Matsunaga T, Sakamaki S, Kuga T, Kuroda H, Kusakabe T, Akiyama T, Konuma Y, Hirayama Y, Kobune M, Kato J, Sasaki K, Kogawa K, Koyama R, Niitsu Y: GST-π gene-transduced hematopoietic progenitor cell transplantation overcomes the bone marrow toxicity of cyclophosphamide in mice. Hum Gene Ther 2000;11:1671–1681.
  140. Jelinek J, Rafferty JA, Cmejla R, Hildinger M, Chinnasamy D, Lashford LS, Ostertag W, Margison GP, Dexter TM, Fairbairn LJ, Baum C: A novel dual function retrovirus expressing multidrug resistance 1 and O6-alkylguanine-DNA-alkyltransferase for engineering resistance of haemopoietic progenitor cells to multiple chemotherapeutic agents. Gene Ther 1999;6:1489–1493.
  141. Fantz CR, Shaw D, Moore JG, Spencer HT: Retroviral coexpression of thymidylate synthase and dihydrofolate reductase confers fluoropyrimidine and antifolate resistance. Biochem Biophys Res Commun 1998;243:6–12.
  142. Donahue RE, Kessler SW, Bodine D, McDonagh K, Dunbar C, Goodman S, Agricola B, Byrne E, Raffeld M, Moen R, Bacher J, Zsebo KM, Nienhuis AW: Helper virus induced T cell lymphoma in nonhuman primates after retroviral mediated gene transfer. J Exp Med 1992;176:1125–1135.
  143. Li Z, Düllmann J, Schiedlmeier B, Schmidt M, von Kalle C, Meyer J, Forster M, Stocking C, Wahlers A, Frank O, Ostertag W, Kühlcke K, Eckert HG, Fehse B, Baum C: Murine leukemia induced by retroviral gene marking. Science 2002;296:497.
  144. Check E: A tragic setback. Nature 2002;420:116–118.
  145. Rill DR, Santana VM, Roberts WM, Nilson T, Bowman LC, Krance RA, Heslop HE, Moen RC, Ihle JN, Brenner MK: Direct demonstration that autologous bone marrow transplantation for solid tumors can return a multiplicity of tumorigenic cells. Blood 1994;84:380–383.

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