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.
Attempts to expand repopulating hematopoietic cells ex vivo have yielded only modest amplification in stem cell numbers. We now report that expression of an exogenous human multi-drug resistance 1 (MDR1) gene enables dramatic ex vivo stem cell expansion in the presence of early acting hematopoietic cytokines. Bone marrow cells were transduced with retroviral vectors expressing either the MDR1 gene or a variant of human dihydrofolate reductase (DHFR), and then expanded for 12 days in the presence of interleukin-3 (IL-3), IL-6, and stem cell factor. When these cells were injected into nonirradiated mice, high levels of long-term engraftment were only seen with MDR1-transduced grafts. To verify that expansion of MDR1-transduced repopulating cells had occurred, competitive repopulation assays were performed using MDR1 expanded grafts. These experiments showed progressive expansion of MDR1-transduced repopulating cells over the expansion period, with a 13-fold overall increase in stem cells after 12 days. In all of the experiments, mice transplanted with expanded MDR1-transduced stem cells developed a myeloproliferative disorder characterized by high peripheral white blood cell counts and splenomegaly. These results show that MDR1-transduced stem cells can be expanded in vitro using hematopoietic cytokines without any drug selection, but enforced stem cell self-renewal divisions can have adverse consequences.
Bunting
K D
KD
Division of Experimental Hematology, the Department of Immunology, and the Department of Biochemistry, St Jude Children's Research Hospital, Memphis, TN, USA.
Galipeau
J
J
Topham
D
D
Benaim
E
E
Sorrentino
B P
BP
D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1 / Q000235:genetics / Q000502:physiology*; D000818:Animals; D002455:Cell Division; D002478:Cells, Cultured / Q000187:drug effects / Q000637:transplantation; D019450:Genes, MDR; D005822:Genetic Vectors; D006085:Graft Survival; D006251:Harvey murine sarcoma virus / Q000235:genetics; D006410:Hematopoiesis / Q000187:drug effects; D018380:Hematopoietic Stem Cell Transplantation / Q000009:adverse effects*; D006801:Humans; D007377:Interleukin-3 / Q000494:pharmacology; D015850:Interleukin-6 / Q000494:pharmacology; D051379:Mice; D008810:Mice, Inbred C57BL; D009196:Myeloproliferative Disorders / Q000209:etiology* / Q000235:genetics / Q000473:pathology; D019089:Stem Cell Factor / Q000494:pharmacology; D014162:Transfection
D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1; D007377:Interleukin-3; D015850:Interleukin-6; D019089:Stem Cell Factor