Construction of antibody/insulin receptor chimera for growth induction of mammalian cells

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• 作者：Hideto Nakabayashi (1)
  Masahiro Kawahara (1)
  Kento Tanaka (1)
  Teruyuki Nagamune (1)
  
• 关键词：Insulin receptor ; Chimeric protein ; Antibody ; Cell growth ; Signal transduction
• 刊名：Cytotechnology
• 出版年：2013
• 出版时间：December 2013
• 年：2013
• 卷：65
• 期：6
• 页码：945-953
• 全文大小：
• 作者单位：Hideto Nakabayashi (1)
  Masahiro Kawahara (1)
  Kento Tanaka (1)
  Teruyuki Nagamune (1)
  
  1. Department of Chemistry and Biotechnology, School of Engineering, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, 113-8656, Tokyo, Japan
  
• ISSN：1573-0778

文摘

The insulin receptor (IR) is expressed ubiquitously in various tissues, where insulin exerts various biological effects on the target cells, such as cellular metabolic changes, cell proliferation and differentiation. Therefore, mimicry of insulin signaling would be a promising strategy to realize artificial control of such cellular fates. In this study, we constructed an antibody/insulin receptor chimera that enables to utilize any antigen as the ligand in principle. We constructed chimeric receptors consisting of anti-fluorescein single chain Fv (scFv), the extracellular D2 domain of erythropoietin receptor and the transmembrane/intracellular domains of IR (scFv-IR; S-IR). The function of S-IR was evaluated in terms of growth signal transduction in murine pro-B Ba/F3 cells and murine fibroblast NIH/3T3 cells. S-IR exerted IL-3-independent cell growth in Ba/F3 cells, while NIH/3T3 cells expressing S-IR acquired growth advantage over parental NIH/3T3 cells in a low-serum condition. S-IR induced phosphorylation of S-IR itself and key signaling molecules downstream of IR. Although antigen-independent activation was significantly observed, S-IR enabled specific amplification of the gene-transduced cells.