|
|
 Previous Article | Table of Contents | Next Article 
Development of a Novel Selective Amplifier Gene for Controllable Expansion of Transduced Hematopoietic Cells
Keiko Ito,
Yasuji Ueda,
Masaki Kokubun,
Masashi Urabe,
Toshiya Inaba,
Hiroyuki Mano,
Hirofumi Hamada,
Toshio Kitamura,
Hideaki Mizoguchi,
Tsuneaki Sakata,
Mamoru Hasegawa, and
Keiya Ozawa
From the Department of Molecular Biology, Institute of Hematology, Jichi Medical School, Tochigi; Department of Hematology, Tokyo Women's Medical College, Tokyo; Laboratory of Molecular Biotherapy, Cancer Chemotherapy Center, Tokyo; Department of Hematopoietic Factors, Institute of Medical Science, The University of Tokyo, Tokyo; and DNAVEC Research Inc, Ibaraki, Japan.
To overcome the low efficiency of gene transfer into hematopoietic cells, we developed a novel system for selective expansion of transduced cells. To this end, we constructed a chimeric cDNA (GCRER) encoding the fusion protein between the granulocyte colony-stimulating factor receptor (G-CSFR) and the hormone-binding domain (HBD) of the estrogen receptor (ER) as a selective amplifier gene. Use of the intracellular signaling pathway of G-CSFR was considered to be appropriate, because G-CSF has the ability not only to stimulate the neutrophil production, but also to expand the hematopoietic stem/progenitor cell pool in vivo. To activate the exogenous G-CSFR signal domain selectively, the estrogen/ER-HBD system was used as a molecular switch in this study. When the GCRER gene was expressed in the interleukin-3 (IL-3)-dependent murine cell line, Ba/F3, the cells showed IL-3-independent growth in response to G-CSF or estrogen. Moreover, the Ba/F3 cells transfected with the  (5-195)GCRER, whose product lacks the extracellular G-CSF-binding domain, did not respond to G-CSF, but retained the ability for estrogen-dependent growth. Further, murine bone marrow cells transduced with the GCRER or (5-195)GCRER gene with retroviral vectors formed a significant number of colonies in response to estrogen, as well as G-CSF, whereas estrogen did not stimulate colony formation by untransduced murine bone marrow cells. It is noteworthy that erythroid colonies were apparently formed by the bone marrow cells transduced with the GCRER gene in the presence of estrogen without the addition of erythropoietin, suggesting that the signals from the G-CSFR portion of the chimeric molecules do not preferentially induce neutrophilic differentiation, but just promote the differentiation depending on the nature of the target cells. We speculate that when the selective amplifier genes are expressed in the primitive hematopoietic stem cells, the growth signal predominates and that the population of transduced stem cells expands upon estrogen treatment, even if some of the cells enter the differentiation pathway. The present study suggests that this strategy is applicable to the in vivo selective expansion of transduced hematopoietic stem cells.
Blood, Vol. 90 No. 10 (November 15), 1997:
pp. 3884-3892
© 1997 by The American Society of Hematology.
 CiteULike  Connotea  Del.icio.us  Digg  Reddit  Technorati What's this?
This article has been cited by other articles:
|
|
|
|
 
M. Kawahara, H. Ueda, K. Tsumoto, I. Kumagai, and T. Nagamune
Mimicry of Erythropoietin and Interleukin-6 Signalling by an Antibody/Cytokine Receptor Chimera in Murine Myeloid 32D Cells
J. Biochem.,
April 1, 2007;
141(4):
563 - 571.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
R. E. Richard and C. A. Blau
Small-Molecule-Directed Mpl Signaling Can Complement Growth Factors to Selectively Expand Genetically Modified Cord Blood Cells
Stem Cells,
January 1, 2003;
21(1):
71 - 78.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
M. C. Walters, A. W. Nienhuis, and E. Vichinsky
Novel Therapeutic Approaches in Sickle Cell Disease
Hematology,
January 1, 2002;
2002(1):
10 - 34.
[Abstract]
[Full Text]
|
|
|
|
|
|
|
T. Neff and C. A. Blau
Pharmacologically regulated cell therapy
Blood,
May 1, 2001;
97(9):
2535 - 2540.
[Full Text]
[PDF]
|
|
|
|
|
|
|
Y. Hanazono, K. Terao, and K. Ozawa
Gene Transfer into Nonhuman Primate Hematopoietic Stem Cells: Implications for Gene Therapy
Stem Cells,
January 1, 2001;
19(1):
12 - 23.
[Abstract]
[Full Text]
|
|
|
|
|
|
|
K. Yoshida, Y. Yamashita, A. Miyazato, K.-i. Ohya, A. Kitanaka, U. Ikeda, K. Shimada, T. Yamanaka, K. Ozawa, and H. Mano
Mediation by the Protein-tyrosine Kinase Tec of Signaling between the B Cell Antigen Receptor and Dok-1
J. Biol. Chem.,
August 4, 2000;
275(32):
24945 - 24952.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
