Abstract
The human multiple drug resistance (MDR) gene has been used as a model for human gene transfer which could lead to human gene therapy. MDR is a transmembrane protein which pumps a number of toxic substances out of cells including several drugs used in cancer chemotherapy. Normal bone marrow cells express low levels of MDR and are particularly sensitive to the toxic effects of these drugs. There are two general applications of MDR gene therapy: (1) to provide drug-resistance to the marrow of cancer patients receiving chemotherapy, and (2) as a selectable marker which when co-transferred with a non-selectable gene such as the human beta globin gene can be used to enrich the marrow for cells containing both genes. We demonstrate efficient transfer and expression of the human MDR gene in a retroviral vector into live mice and human marrow cells including CD34+ cells isolated from marrow and containing the bulk of human hematopoietic progenitors. MDR gene transduction corrects the sensitivity of CD34+ cells to taxol, an MDR drug substrate, and enriches the marrow for MDR-transduced cells. The MDR gene-containing retroviral supernatant used has been shown to be safe and free of replication-competent retrovirus. Because of the safety of the MDR retroviral supernatant, and efficient gene transfer into mouse and human marrow cells, a phase 1 clinical protocol for MDR gene transfer into cancer patients has been approved to evaluate MDR gene transfer and expression in human marrow.
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References
Aihara M, Aihara Y, Schmidt-Wolf G, Schmidt-Wolf I, Sikic BI, Blume KG & Chao NJ (1991) A combined approach for purging multidrug-resistant leukemic cell lines in bone marrow using a monoclonal antibody and chemotherapy. Blood 77: 2079–2084.
Berenson RJ (1992) Transplantation of CD34+ hematopoietic precursors: clinical rationale. [Review]. Transplant Proc. 24: 3032–3034.
Berenson RJ, Bensinger WI, Kalamasz DF, Heimfeld S, Goffe RA, Berninger RW, Peterson DR, Thompson P & Strong DM (1992) Transplantation of stem cells enriched by immunoadsorption. Prog. Clin. Biol. Res. 377: 449–457.
Eaves CJ, Sutherland HF, Udomsakdi C, Lansdorp PM, Szilvassy SJ, Fraser CC, Humphries RK, Barnett MJ, Phillips GL & Eaves AC (1992) The human hematopoietic stem cellin vitro andin vivo — see comments—. Blood Cells 18: 301–307.
Galski H, Sullivan M, Willingham MC, Chim KV, Gottesman MM, Pastan I & Merlino GT (1989) Expression of a human multidrug resistance cDNA (MDR1) in the bone marrow of transgenic mice: resistance to daunomycin-induced leukopenia. Mol. Cell. Biol. 9: 4357–4363.
Hesdorffer C & Antman K (1994) Clinical protocol: Human MDR gene transfer in patients with advanced cancer. Human Gene Therapy 5: 1151–1160.
Hesdorffer C, Ward M, Markowitz D & Bank A (1990) Efficient gene transfer in live mice using a unique retroviral packaging line. DNA & Cell Biol. 9: 717–723.
Kessinger A & Armitage JO (1991) The evolving role of autologous peripheral stem cell transplantation following high-dose therapy for malignancies. [Editorial] Blood 77: 211–213.
Kessinger A, Armitage JO, Landmark JD, Smith DM & Weisenburger DD (1988) Autologous peripheral hematopoietic stem cell transplantation restores hematopoietic function following marrow ablative therapy. Blood 71: 723–727.
Lebkowski JS, Schain LR, Okrongly D, Levinsky R, Harvey MJ & Okarma TB (1992) Rapid isolation of human CD34 hematopoietic stem cells — purging of human tumor cells. Transplantation 53: 1011–1019.
Lerner N, Brigham S, Goff S & Bank A (1987) Human beta globin gene expression after gene transfer using retroviral vectors. DNA 6: 573–581.
Markowitz D, Goff S & Bank A (1988a) Construction and use of a safe and efficient amphotropic packaging cell line. Virology 167: 400–405.
Markowitz D, Goff S & Bank A (1988b) A safe packaging line for gene transfer: Separating viral genes on two different plasmids. J. Virol. 62: 1120–1125.
Mickisch GH, Aksentijevich I, Schoenlein PV, Goldstein LJ, Galski H, Stahle C, Sachs DH, Pastan I & Gottesman MM (1992) Trans-plantation of bone marrow cells from transgenic mice expressing the human MDR1 gene results in long-term protection against the myelosuppressive effect of chemotherapy in mice. Blood 79: 1087–1093.
Miller A (1990) Progress toward human gene therapy. Blood 76: 271–278.
Moore MA (1991) Review: Stratton Lecture 1990. Clinical implications of positive and negative hematopoietic stem cell regulators. [Review]. Blood 78: 1–19.
Noonan KE, Beck C, Holzmayer TA, Chin JE, Wunder JS, Andrulis IL, Gazdar AF, Willman CL, Griffith B, Von Hoff DD & Roninson I (1990) Quantitative analysis of MDR1 (multidrug resistance) gene expression in human tumors by polymerase chain reaction. Proc. Natl. Acad. Sci. USA 87: 7160–7164.
Okarma T, Lebkowski J, Schain L, Harvey M, Tricot G, Srour E, Meyers WG, Burnett A, Sniecinski I & O'Reilly RJ (1992) The AIS Selector: A new technology for stem cell purification. In: Adv. in Bone Marrow Purging and Processing (pp. 487–504) Wiley-Liss, New York.
Pastan I & Gottesman MM (1991). Multidrug resistance. Annu. Rev. Med. 42: 277–286.
Podda S, Ward M, Himelstein A, Richardson C, De la Flor-Weiss E, Smith E, Gottesman M, Pastan I & Bank A (1992) Transfer and expression of the human multiple drug resistance gene into live mice. Proc. Natl. Acad. Sci. USA 89: 9676–9680.
Richardson C, Ward M & Bank A (1993) Mouse fetal liver cells lack functional receptors for amphotropic retroviruses. Blood Suppl. 82: 2200A.
Sutherland HJ, Eaves C, Dragowska W et al. (1989) Characterization and partial purification of human marrow cells capable of initiating long-term hematopoiesisin vitro. Blood 74: 1563–1570.
Ward M, Richardson C, Pioli P, Aiello J, Reiss R, Hesdorffer C & Bank A (1994a) MDR gene transfer and expression in peripheral blood progenitors. Blood 85: 356a (Abstract).
Ward M, Richardson C, Pioli P, Smith L, Podda S, Goff S, Hesdorffer C & Bank A (1994b) Transfer and expression of the human multiple drug resistance gene in human CD34+ cells. Blood 84: 1408–1414.
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Bank, A., Ward, M., Richardson, C. et al. Transfer and expression of the human multiple drug resistance gene as potential human gene therapy. Cytotechnology 18, 119–124 (1995). https://doi.org/10.1007/BF00744327
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DOI: https://doi.org/10.1007/BF00744327