2007 Fiscal Year Final Research Report Summary
Insulin-like growth factor-I receptor-targeted cancer therapy
| Project/Area Number |
17570120
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Functional biochemistry
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| Research Institution | Tokai University |
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Principal Investigator |
FUJITA Yoko Tokai University, School of Engineering, Professor (10328106)
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| Co-Investigator(Kenkyū-buntansha) |
TAKAYANAGI Atsushi Keio University, School of Medicine, Junior Associate Professor (80245464)
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| Project Period (FY) |
2005 – 2007
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| Keywords | Anti-IGF-I receptor / single chain antibody / cancer molecular thera / downregulation / Anti-tumor erowth |
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| Research Abstract |
The aims of this study were to produce more effective therapeutic antihodies against the IGF-I receptor which can inhibit cancer growth, and to understand mechanisms by which anti-IGFIR induce down-regulation of IGFIR.
1. Preparation of human anti-IGFIR with higher affinities than that of the 1H7 single chain antibody (scFv).
We obtained 20 clones from a phage library displaying human scFvs. After extensive analyses of those clones, however, we found that they were cross-reactive to the insulin receptor as well as some other non-related proteins. In contrast, scFvs prepared from hybridoma producing 1H7 or 3B7 anti-IGFIR monoclonal antibody (mAb) showed strict specificity for IGFIR. These results suggested that rigorous screening is required to obtain scFvs of our interest in vitro antibody production using phage display technologies. Construction, expression, purification, and characterization of the two scFvs derived from hybridomas resulted in valuable information on their affinity and epitopes. The results have been published.
2. Understanding mechanism by which anti-IGFIR antibodies induce down-regulation of IGFIR
We found that all five mAbs used, despite showing different effects on IGFIR signaling, down-regulated IGFIR. In contrast, IGF-I apparently internalized the receptor, but did not lead to its degradation as evidenced by intact IGFIR subunits detected by immunoblotting. Pretreatment of MCF-7 cells with methylamine substantially reduced the antibody-mediated IGFIR down-regulation whereas MG115 did not. No ubiquitination was detected with IGFIR in MCF-7 cells treated by IGFIR mAb nor IGF-I. These results suggest that anti-IGFIR antibodies with different epitope-specificities can cause IGFIR down-regulation via a lysosome-dependent pathway in an IGFIR activation-independent manner. The results were presented at the Annual Meeting of ASBMB, 2007 (manuscript in preparation).
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[Journal Article] Construction and characterization of single-chain antibodies against human insulin-like growth factor-1 Receptor from hybridomas producing 1H7 and 3B7 monoclonal antibody2008
Author(s)
Kusada, Y, Morizono, T., Matsumoto-Takasaki, A., Sakai, K., Sato, S., Asanuma, H., Takayanagi, A., and Fujita-Yamaguchi, Y.
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Journal Title
J.Biochem. 143
Pages: 9-19
Description
「研究成果報告書概要(和文)」より
Peer Reviewed
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[Journal Article] Construction and characterization of single-chain antibodies against human insulin-like growth factor-1 Receptor from hybridomas producing 1H7 and 3B7 monoclonal antibody2008
Author(s)
Kusada, Y, Morizono, T., Matsumoto-Takasaki, A., Sakai, K., Sato, S., Asanuma, H., Takayanagi, A., Fujita-Yamaguchi, Y
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Journal Title
J. Biochem 143
Pages: 9-19
Description
「研究成果報告書概要(欧文)」より
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[Presentation] Isolation and characterization of human single-chain antibodies against insulin receptor isoform A by phage-display techniques2007
Author(s)
Kusada, Y., Sekine, K., Takayanagi, A., Shimizu, N., and Fujita-Yamaguchi, Y.
Organizer
11th Annual Meeting of Japanese Society for Molecular Targeted Therapy for Cancer
Place of Presentation
Osaka,Japan
Year and Date
20070706-07
Description
「研究成果報告書概要(和文)」より
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[Presentation] Isolation and characterization of human single-chain antibodies against insulin receptor isoform A by phage-display techniques2007
Author(s)
Kusada, Y, Sekine, K, Takayanagi, A., Shimizu, N., Fujita-Yamaguchi, Y
Organizer
11th Annual Meeting of Japanese Society for Molecular Targeted Therapy for Cancer
Place of Presentation
Osaka, Japan
Year and Date
20070706-07
Description
「研究成果報告書概要(欧文)」より
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[Presentation] Comparison of monoclonal antibodies against human IGF-I receptor prepared from mice to those prepared by phage-display technology in vitro2007
Author(s)
Kusada, Y., Sekine, K., Sato, S., Asanuma, H., Takayanagi, A., Shimizu, N., and Fujita-Yamaguchi, Y.
Organizer
WIRC
Place of Presentation
Davos,Switzerland
Year and Date
20070411-14
Description
「研究成果報告書概要(和文)」より
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[Presentation] Comparison of monoclonal antibodies against human IGF-I receptor prepared from mice to those prepared by phage-display technology in vitro2007
Author(s)
Kusada, Y., Sekine, K., Sato, S., Asanuma, H., Takayanagi, A., Shimizu, N., Fujita-Yamaguchi, Y
Place of Presentation
WIRC, Davos, Switzerland
Year and Date
20070411-14
Description
「研究成果報告書概要(欧文)」より
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[Presentation] Isolation and characterization of single-chain antibodies(scFvs)against insulin-like growth factor-I receptor(IGF-IR)and/or insulin receptor isoform A(IR-A)2006
Author(s)
Kusada, Y., Sekine, H., Ohtani, M., Sakai, K., Yajima, Y., Takayanagi, A., Shimizu, N., and Fujita-Yamaguchi, Y.
Organizer
20th IUBMB International Congress of Biochemistry and Molecular Biology and 11th FAOBMB Congress
Place of Presentation
Kyoto,Japan
Year and Date
20060618-23
Description
「研究成果報告書概要(和文)」より
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[Presentation] Isolation and characterization of single-chain antibodies(scFvs) against insulin-like growth factor-I receptor(IGF-IR) and/or insulin receptor isoform A(IR-A)2006
Author(s)
Kusada, Y., Sekine, H., Ohtani, M., Sakai, K., Yajima, Y., Takayanagi, A., Shimizu, N., Fujita-Yamaguchi, Y
Organizer
20th IUBMB International Congress of Biochemistry and Molecular Biology and 11th FAOBMB Congress
Place of Presentation
Kyoto, Japan
Year and Date
20060618-23
Description
「研究成果報告書概要(欧文)」より
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[Presentation] Isolation and characterization of human single-chain antibodies(scFv)against insulin-like growth factor I receptor(IGF-IR)2005
Author(s)
Kusada, Y., Morizono, T., Sakai, K., Takayanagi, A., Shimizu, N., and Fujita-Yamaguchi, Y.
Organizer
78th Annual Meeting of the Japanese Biochemical Society
Place of Presentation
Kobe,Japan.
Year and Date
20051019-22
Description
「研究成果報告書概要(和文)」より
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[Presentation] Isolation and characterization of human single-chain antibodies(scFv) against insulin-like growth factor I receptor(IGF-IR)2005
Author(s)
Kusada, Y., Morizono, T., Sakai, K., Takayanagi, A., Shimizu, N., Fujita-Yamaguchi, Y
Organizer
78th Annual Meeting of the Japanese Biochemical Society
Place of Presentation
Kobe, Japan
Year and Date
20051019-22
Description
「研究成果報告書概要(欧文)」より
