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Transfer and expression of the human multiple drug resistance gene in human
CD34+ cells
M Ward, C Richardson, P Pioli, L Smith, S Podda, S Goff, C Hesdorffer and A Bank
Department of Medicine, Columbia University New York, NY 10032.
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.
Volume 84,
Issue 5,
pp. 1408-1414,
09/01/1994
Copyright © 1994 by The American Society of Hematology
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