title: 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.
abstract: The MDR1 multidrug resistance gene confers resistance to natural-product anticancer drugs including paclitaxel. We conducted a clinical gene therapy study to determine whether retroviral-mediated transfer of MDR1 in human hematopoietic cells would result in stable engraftment, and possibly expansion, of cells containing this gene after treatment with myelosuppressive doses of paclitaxel. Patients with metastatic breast cancer who achieved a complete or partial remission after standard chemotherapy were eligible for the study. Hematopoietic stem cells (HSCs) were collected by both peripheral blood apheresis and bone marrow harvest after mobilization with a single dose of cyclophosphamide (4 g/m2) and daily filgrastim therapy (10 microg/kg/day). After enrichment for CD34+ cells, one-third of each collection was incubated ex vivo for 72 h with a replication-incompetent retrovirus containing the MDR1 gene (G1MD) in the presence of stem-cell factor, interleukin 3, and interleukin 6. The remaining CD34+ cells were stored without further manipulation. All of the CD34+ cells were reinfused for hematopoietic rescue after conditioning chemotherapy with ifosfamide, carboplatin, and etoposide regimen. After hematopoietic recovery, patients received six cycles of paclitaxel (175 mg/m2 every 3 weeks). Bone marrow and serial peripheral blood samples were obtained and tested for the presence of the MDR1 transgene using a PCR assay. Six patients were enrolled in the study and four patients received infusion of genetically altered cells. The ex vivo transduction efficiency, estimated by the PCR assay, ranged from 0.1 to 0.5%. Three of the four patients demonstrated engraftment of cells containing the MDR1 transgene. The estimated percentage of granulocytes containing the MDR1 transgene ranged from a maximum of 9% of circulating nucleated cells down to the limit of detection of 0.01%. One patient remained positive for the MDR1 transgene throughout all six cycles of paclitaxel therapy, whereas the other 2 patients showed a decrease in the number of cells containing the transgene to undetectable levels. Despite the low level of engraftment of MDR1-marked cells, a correlation was observed between the relative number of granulocytes containing the MDR1 transgene and the granulocyte nadir after paclitaxel therapy. No adverse reactions to the genetic manipulation procedures were detected. Therefore, engraftment of human HSCs transduced with the MDR1 gene can be achieved. However, the overall transduction efficiency and stable engraftment of gene-modified HSCs must be improved before MDR1 gene therapy and in vivo selection with anticancer drugs can be reliably used to protect cancer patients from drug-related myelosuppression.
authors.affiliation: National Cancer Institute, National Institutes of Health, Bethesda, Maryland 20892, USA. kcowan@unmc.edu
authors.forename: K H
authors.initials: KH
authors.lastname: Cowan
authors.forename: J A
authors.initials: JA
authors.lastname: Moscow
authors.forename: H
authors.initials: H
authors.lastname: Huang
authors.forename: J A
authors.initials: JA
authors.lastname: Zujewski
authors.forename: J
authors.initials: J
authors.lastname: O'Shaughnessy
authors.forename: B
authors.initials: B
authors.lastname: Sorrentino
authors.forename: K
authors.initials: K
authors.lastname: Hines
authors.forename: C
authors.initials: C
authors.lastname: Carter
authors.forename: E
authors.initials: E
authors.lastname: Schneider
authors.forename: G
authors.initials: G
authors.lastname: Cusack
authors.forename: M
authors.initials: M
authors.lastname: Noone
authors.forename: C
authors.initials: C
authors.lastname: Dunbar
authors.forename: S
authors.initials: S
authors.lastname: Steinberg
authors.forename: W
authors.initials: W
authors.lastname: Wilson
authors.forename: B
authors.initials: B
authors.lastname: Goldspiel
authors.forename: E J
authors.initials: EJ
authors.lastname: Read
authors.forename: S F
authors.initials: SF
authors.lastname: Leitman
authors.forename: K
authors.initials: K
authors.lastname: McDonagh
authors.forename: C
authors.initials: C
authors.lastname: Chow
authors.forename: A
authors.initials: A
authors.lastname: Abati
authors.forename: Y
authors.initials: Y
authors.lastname: Chiang
authors.forename: Y N
authors.initials: YN
authors.lastname: Chang
authors.forename: M M
authors.initials: MM
authors.lastname: Gottesman
authors.forename: I
authors.initials: I
authors.lastname: Pastan
authors.forename: A
authors.initials: A
authors.lastname: Nienhuis
mesh_terms: D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1 / Q000235:genetics*; D000328:Adult; D018952:Antigens, CD34 / Q000032:analysis; D000972:Antineoplastic Agents, Phytogenic / Q000009:adverse effects / Q000627:therapeutic use; D001943:Breast Neoplasms / Q000188:drug therapy / Q000628:therapy*; D003131:Combined Modality Therapy; D018076:DNA, Complementary / Q000235:genetics; D064420:Drug-Related Side Effects and Adverse Reactions / Q000517:prevention & control; D005260:Female; D015316:Genetic Therapy; D005822:Genetic Vectors; D018380:Hematopoietic Stem Cell Transplantation; D006801:Humans; D008875:Middle Aged; D017239:Paclitaxel / Q000009:adverse effects / Q000627:therapeutic use*; D010865:Pilot Projects; D016133:Polymerase Chain Reaction; D012190:Retroviridae / Q000235:genetics; D016176:T-Lymphocyte Subsets; D014161:Transduction, Genetic; D014182:Transplantation, Autologous
chemical_list: D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1; D018952:Antigens, CD34; D000972:Antineoplastic Agents, Phytogenic; D018076:DNA, Complementary; D017239:Paclitaxel