Aug 24, 1999

Development of a modified selective amplifier gene for hematopoietic stem cell gene therapy

Gene Therapy
K M MatsudaK Ozawa

Abstract

We have proposed a novel concept, ie selective expansion of transduced cells, to overcome the low efficiency of gene transfer into hematopoietic stem cells. Previously, a fusion gene encoding a chimeric receptor (DeltaGCRER) between the mouse granulocyte colony-stimulating factor receptor (G-CSFR) and the hormone-binding domain of rat estrogen receptor was constructed as a 'selective amplifier gene'. Although the chimeric gene conferred estrogen-inducible proliferation on the transduced Ba/F3 cells, it also mediated differentiation of the retrovirally transduced 32D cells upon estrogen treatment. Since only a growth signal is required for our purpose, we further modified the DeltaGCRER gene to attenuate its differentiation signal. Based on the observation that tyrosine-703 in wild-type G-CSFR plays a pivotal role in transmitting the differentiation signal, phenylalanine was substituted for this residue in DeltaGCRER. When the resultant selective amplifier gene (DeltaY703F-GCRER gene) was expressed in 32D cells, sustained growth was supported by estrogen, while differentiation was suppressed. These cells ceased to grow upon estrogen withdrawal and differentiated with G-CSF treatment. The present findings suggested that DeltaY703...Continue Reading

Mentioned in this Paper

Filgrastim
Fusion Gene
Gene Transfer Techniques
Colony-Forming Units, Hematopoietic
Gene Amplification
Estrogen receptor alpha, human
Granulocyte Colony-Stimulating Factor
Chimera Organism
Cell Communication
Cell Differentiation Process

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