Transfer and expression of the human multiple drug resistance gene in human CD34+ cells

Blood. 1994 Sep 1;84(5):1408-14.

Abstract

The human multiple-drug resistance (MDR1) gene has been transferred into human hematopoietic progenitors using retroviral gene transfer. Human bone marrow cells and isolated CD34+ cells isolated from marrow were exposed to growth factors interleukin-3 (IL-3), IL-6, and stem cell factor for 48 hours and then to two changes of MDR retroviral supernatants over the next 24 hours. Progenitor assays in methylcellulose at this time showed that 18% to 70% of BFU-E and 30% to 60% of CFU-GM contain the transferred MDR gene by polymerase chain reaction analysis. Up to 11.2% of the progeny of these cells express increased amounts of MDR glycoprotein on their surface by fluorescence-activated cell sorter (FACS) analysis. In addition, transduced cells are enriched in high MDR-expressing cells after exposure to taxol as assessed by FACS analysis, and by resistance of BFU-E to taxol (Bristol-Myers Squibb, Princeton, NJ). These studies indicate the feasibility of using MDR gene transfer as a means of enriching marrow for MDR-transduced cells. They also provide the basis of a phase 1 clinical protocol in patients with advanced cancers not involving the bone marrow for the use of MDR gene transfer as a means of protecting marrow cells, which normally express low levels of MDR, from the myelosuppressive effects of drugs like taxol.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Antigens, CD / analysis*
  • Antigens, CD34
  • Bone Marrow Cells
  • Colony-Forming Units Assay
  • Drug Resistance / genetics*
  • Flow Cytometry
  • Gene Expression
  • Gene Transfer Techniques
  • Harvey murine sarcoma virus
  • Hematopoietic Cell Growth Factors / pharmacology
  • Hematopoietic Stem Cells / cytology
  • Hematopoietic Stem Cells / drug effects
  • Hematopoietic Stem Cells / metabolism*
  • Humans
  • Interleukin-3 / pharmacology
  • Interleukin-6 / pharmacology
  • Paclitaxel / toxicity*
  • Polymerase Chain Reaction
  • Stem Cell Factor
  • Transduction, Genetic
  • Transfection

Substances

  • Antigens, CD
  • Antigens, CD34
  • Hematopoietic Cell Growth Factors
  • Interleukin-3
  • Interleukin-6
  • Stem Cell Factor
  • Paclitaxel