Selection and growth regulation of genetically modified cells with hapten-specific antibody/receptor tyrosine kinase chimera.
Although receptor tyrosine kinases (RTKs) play a pivotal role in the development and maintaining the homeostasis of the body, overexpression or mutation of RTKs often induces tumorigenesis or metastasis. To mimic the function of RTKs, we developed two fusion receptors consisting of anti-fluorescein antibody single-chain Fv, extracellular D2 domain of erythropoietin receptor and transmembrane/intracellular domains of epidermal growth factor receptor or c-fms based on previously constructed antibody/cytokine receptor chimeras. The expression of these chimeric receptors in the hematopoietic cell line Ba/F3 and non-hematopoietic cell line NIH/3T3 resulted in the activation of receptors themselves, downstream signaling molecules and cell proliferation in response to fluorescein-conjugated BSA, leading to selective expansion of transduced cells up to almost 100%. These results indicate that the cognate antigen could activate the chimeric receptors even though the wild-type extracellular domains were switched to the antibody fragment. This is the first study to show that our antigen-mediated genetically modified cell amplification (AMEGA) system could be applied to non-hematopoietic cells by utilizing antibody/RTK chimeras.
Tanaka
Kento
K
Department of Chemistry and Biotechnology, School of Engineering, University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-8656, Japan.
Kawahara
Masahiro
M
Ueda
Hiroshi
H
Nagamune
Teruyuki
T
D000818:Animals; D002460:Cell Line; D049109:Cell Proliferation; D006241:Haptens / Q000378:metabolism; D006412:Hematopoietic Stem Cells / Q000166:cytology / Q000378:metabolism; D007135:Immunoglobulin Variable Region / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D015202:Protein Engineering; D020794:Receptor Protein-Tyrosine Kinases / Q000235:genetics / Q000378:metabolism*; D017467:Receptors, Erythropoietin / Q000235:genetics / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction
D006241:Haptens; D007135:Immunoglobulin Variable Region; D017467:Receptors, Erythropoietin; D011993:Recombinant Fusion Proteins; D020794:Receptor Protein-Tyrosine Kinases