Development of a novel selective amplifier gene for controllable expansion of transduced hematopoietic cells.
Development of a novel selective amplifier gene for controllable expansion of transduced hematopoietic cells.
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开发一种新型选择性扩增基因，用于可控扩增转导的造血细胞。
DOI：
10.1182/blood.v90.10.3884
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发表时间：
1997
期刊：
影响因子：
20.3
通讯作者：
Keiya Ozawa
中科院分区：
文献类型：
--
作者：
Keiko Ito;Y. Ueda;Masaki Kokubun;M. Urabe;Toshiya Inaba;Hiroyuki Mano;Hirofumi Hamada;Toshio Kitamura;Hideaki Mizoguchi;Tsuneaki Tsukuba;Mamoru Hasegawa;Keiya Ozawa
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 Delta(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 Delta(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.
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用于检测逆转录病毒感染的新报告系统。
影响因子：
5.1
作者：
Planelles,V;Haislip,A;Withers-Ward,ES;Stewart,SA;Xie,Y;Shah,NP;Chen,IS
通讯作者：
Chen,IS
影响因子：
20.3
作者：
Jay Tong;M. Gordon;E. Srour;R. Cooper;A. Orazi;I. Mcniece;R. Hoffman
通讯作者：
Jay Tong;M. Gordon;E. Srour;R. Cooper;A. Orazi;I. Mcniece;R. Hoffman
逆转录病毒介导的表达克隆的应用。
DOI：
--
发表时间：
1996
期刊：
Experimental hematology.
影响因子：
--
作者：
Onishi,M;Kinoshita,S;Morikawa,Y;Shibuya,A;Phillips,J;Lanier,LL;Gorman,DM;Nolan,GP;Miyajima,A;Kitamura,T
通讯作者：
Kitamura,T
DOI：
10.1073/pnas.83.8.2496
发表时间：
1986-04-01
影响因子：
11.1
作者：
BERTHOIS, Y;KATZENELLENBOGEN, JA;KATZENELLENBOGEN, BS
通讯作者：
KATZENELLENBOGEN, BS
连续移植表明，在小鼠基因治疗模型中，早期造血前体细胞被 MDR-1 逆转录病毒载体转导。
影响因子：
6.4
作者：
Hanania,EG;Deisseroth,AB
通讯作者：
Deisseroth,AB