RRC ID 39327
Author Xu R, Kume A, Matsuda KM, Ueda Y, Kodaira H, Ogasawara Y, Urabe M, Kato I, Hasegawa M, Ozawa K.
Title A selective amplifier gene for tamoxifen-inducible expansion of hematopoietic cells.
Journal J Gene Med
Abstract BACKGROUND:We have developed a novel system for expansion of gene-modified hematopoietic stem/progenitor cells to overcome the low efficiency of current gene transfer methodology. This system involves 'selective amplifier genes', that encode fusion proteins between the granulocyte colony-stimulating factor receptor (GCR) and the hormone-binding domain of estrogen receptor (ER). Hematopoietic progenitors expressing the chimeras showed estrogen-responsive growth in a controllable manner. However, endogenous estrogen may activate the fusion proteins in vivo, depending on the hormonal status of the subjects.
METHODS:We replaced ER with a mutant receptor (TmR) which specifically binds to 4-hydroxytamoxifen (Tm), to overcome limitations with wild-type ER. Interleukin-3 (IL-3)-dependent Ba/F3 cells and hematopoietic progenitor cells transduced with the resultant fusion proteins (GCRTmR and delta GCRTmR) were examined for ligand-inducible growth.
RESULTS:GCRTmR- and delta GCRTmR-expressing Ba/F3 showed IL-3-independent growth in response to Tm, while the cells were unresponsive to estrogen at concentrations up to 10(-7)-10(-6) M. Furthermore, murine bone marrow cells transduced with GCRTmR and delta GCRTmR formed colonies in methyl-cellulose medium in response to Tm, while virtually no colonies appeared with 10(-7) M estrogen or without cytokines.
CONCLUSIONS:These results suggest that influences of the endogenous estrogen can be almost eliminated by using the GCRTmR/Tm or delta GCRTmR/Tm system to expand gene-modified hematopoietic stem/progenitor cells.
Volume 1(4)
Pages 236-44
Published 1999-1-1
DOI 10.1002/(SICI)1521-2254(199907/08)1:4<236::AID-JGM42>3.0.CO;2-2
PMID 10738556
MeSH Animals Base Sequence Cell Division / drug effects Cell Line Chimera / genetics Colony-Forming Units Assay DNA Primers / genetics Estradiol / pharmacology Gene Amplification* Genetic Therapy Genetic Vectors Hematopoietic Stem Cell Transplantation Hematopoietic Stem Cells / cytology Hematopoietic Stem Cells / drug effects* Hematopoietic Stem Cells / metabolism Mice Receptors, Estrogen / genetics Receptors, Estrogen / metabolism Receptors, Granulocyte Colony-Stimulating Factor / genetics Receptors, Granulocyte Colony-Stimulating Factor / metabolism Recombinant Fusion Proteins / genetics Recombinant Fusion Proteins / metabolism Retroviridae / genetics Tamoxifen / pharmacology* Transduction, Genetic
IF 3.258
Times Cited 23
Human and Animal Cells 10T1/2(RCB0247)