Discovery of protein phosphorylation markers to predict the efficacy of anti-EGFR antibody drugs in colorectal cancer patients

• Project/Area Number: 24790184
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Multi-year Fund
• Research Field: Medical pharmacy
• Research Institution: 独立行政法人医薬基盤研究所
• Principal Investigator: KUGA Takahisa 独立行政法人医薬基盤研究所, 創薬基盤研究部, 研究員 (20551857)
• Research Collaborator: TOMONAGA Takeshi 独立行政法人医薬基盤研究所, 創薬基盤研究部, プロジェクトリーダー (80227644)
• Project Period (FY): 2012-04-01 – 2014-03-31
• Project Status: Completed (Fiscal Year 2013)
• Budget Amount: ¥4,550,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥1,050,000); Fiscal Year 2013: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000); Fiscal Year 2012: ¥2,730,000 (Direct Cost: ¥2,100,000、Indirect Cost: ¥630,000)
• Keywords: オーダーメード医療 / 大腸がん / セツキシマブ / EGFR / キナーゼ / プロテオーム / バイオマーカー / 接着阻止 / 大腸癌 / タンパク質リン酸化 / 細胞接着阻止

Research Abstract

Anti-EGFR antibody drugs have been approved for metastatic colorectal cancer (mCRC) therapy, but the drugs are ineffective for CRC exhibiting EGF-independent activation of the EGFR signaling pathway. Although a part of the patients with the aberrant EGFR signaling can be detected by the KRAS mutation testing, further improvement of the method detecting the EGFR signaling-aberrant patients is needed. In this study, using colorectal cancer cell lines, we examined whether comprehensive analysis of activation levels of kinases related to the EGFR signaling could potentiate to predict the efficacy of the anti-EGFR drugs. In addition, I found a novel mechanism by which the anti-EGFR drugs inhibit proliferation of CRC cells. This study showed several biomarker candidates to predict the efficiency of the anti-EGFR antibody drugs.

Research Products

• [Journal Article] Increased expression of sorcin is associated with multidrug resistance in leukemia cells via up-regulation of MDR1 expression through cAMP response element-binding protein (2014)
  • Author(s): Yamagishi N., Nakao R., Kondo R., Nishitsuji M., Saito Y., Kuga T., Hatayama T., and Nakayama Y
  • Journal Title: Biochem Biophys Res Commun Volume: 13 Pages: 430-436
• [Journal Article] Discovery of colorectal cancer biomarker candidates by membrane proteomic analysisand subsequent verification using selected reaction monitoring and tissue microarrayanalysis (2014)
  • Author(s): Kume H, Muraoka S, Kuga T, Adachi J, Narumi R, Watanabe S, Kuwano M, Kodera Y, Matsushita K, Fukuoka J, Masuda T, Ishihama Y, Matsubara H, Nomura F, Tomonaga T.
  • Journal Title: Mol Cell Proteomics. Volume: M113 Issue: 6 Pages: 37093-37093
  • DOI: 10.1074/mcp.m113.037093
  • Peer Reviewed
• [Journal Article] Lamin B2 prevents chromosome instability by ensuring proper mitotic chromosome segregation (2014)
  • Author(s): Kuga T., Nie H., Kazami T., Satoh M., Matsushita K., Nomura F., Maeshima K., Nakayama Y., and Tomonaga T
  • Journal Title: Oncogenesis Volume: 3
• [Journal Article] Genistein induces cytokinesis failure through RhoA delocalization and anaphase chromosome bridging (2014)
  • Author(s): Nakayama Y., Saito Y., Soeda S., Iwamoto E., Ogawa S., Yamagishi N., Kuga T., and Yamaguchi N
  • Journal Title: J Cell Biochem Volume: 115 Pages: 763-771
• [Journal Article] Genistein induces cytokinesis failure through RhoA delocalization and anaphase chromosome bridging. (2014)
  • Author(s): Nakayama Y, Saito Y, Soeda S, Iwamoto E, Ogawa S, Yamagishi N, Kuga T, Yamaguchi N
  • Journal Title: J Cell Biochem. Volume: 115 Issue: 4 Pages: 763-771
  • DOI: 10.1002/jcb.24720
  • Peer Reviewed
• [Journal Article] A novel mechanism of keratin cytoskeleton organization through casein kinase I alpha and FAM83H in colorectal cancer (2013)
  • Author(s): Kuga T., Kume H., Kawasaki N., Sato M., Adachi J., Shiromizu T., Hoshino I., Nishimori T., Matsubara H., and Tomonaga T
  • Journal Title: J Cell Sci Volume: 126 Pages: 4721-4731
• [Journal Article] Identification of Missing Proteins in the neXtProt Database and Unregistered Phosphopeptides in the PhosphoSitePlus Database As Part of the Chromosome-Centric Human Proteome Project (2013)
  • Author(s): Shiromizu T., Adachi J., Watanabe S., Murakami T., Kuga T., Muraoka S., and Tomonaga T
  • Journal Title: J Proteome Res Volume: 12 Pages: 2414-2421
• [Journal Article] Formation of long and winding nuclear F-actin bundles by nuclear c-Abl tyrosine kinase (2013)
  • Author(s): Aoyama K., Yuki R., Horiike Y., Kubota S., Yamaguchi N., Morii M., Ishibashi K., Nakayama Y., Kuga T., Hashimoto Y., Tomonaga T., and Yamaguchi N.
  • Journal Title: Exp Cell Res Volume: 319 Pages: 3251-3268
• [Journal Article] Phosphorylation of KRAB-associated Protein 1 (KAP1) at Tyr-449, Tyr-458, and Tyr-517 by Nuclear Tyrosine Kinases Inhibits the Association of KAP1 and Heterochromatin Protein 1alpha (HP1alpha) with Heterochromatin (2013)
  • Author(s): Kubota S., Fukumoto Y., Aoyama K., Ishibashi K., Yuki R., Morinaga T., Honda T., Yamaguchi N., Hashimoto Y., Kuga T., Tomonaga T., and Yamaguchi N.
  • Journal Title: J Biol Chem Volume: 288 Pages: 17871-17883
• [Journal Article] Serum fibrinogen alpha C-chain 5.9 kDa fragment as a biomarker for early detection of hepatic fibrosis related to hepatitis C virus (2013)
  • Author(s): Sogawa K., Noda K., Umemura H., Seimiya M., Kuga T., Tomonaga T., Nishimura M., Kanai F., Imazeki F., Takizawa H., Yoneda M., Nakajima A., Tsutsumi M., Yokosuka O., and Nomura F.
  • Journal Title: Proteomics Clin Appl Volume: 7 Pages: 424-431
• [Journal Article] Lamin B2 prevents chromosome instability by ensuring proper mitotic chromosome segregation (2013)
  • Author(s): Kuga, T., Nie, H., Kazami, T., Satoh, M., Matsushita, K., Nomura, F., Maeshima, K., Nakayama, Y., Tomonaga, T.
  • Journal Title: Oncogenesis Volume: 3 Issue: 3 Pages: e94-e94
  • DOI: 10.1038/oncsis.2014.6
  • Peer Reviewed
• [Journal Article] A novel mechanism of keratin cytoskeleton organization through casein kinase I alpha and FAM83H in colorectal cancer. (2013)
  • Author(s): Kuga, T., Kume, H., Kawasaki, N., Sato, M., Adachi, J., Shiromizu, T., Hoshino, I., Nishimori, T., Matsubara, H., and Tomonaga T.
  • Journal Title: J Cell Sci Volume: 126 Pages: 4721-31
  • DOI: 10.1242/jcs.129684
  • Peer Reviewed
• [Journal Article] Formation of long and winding nuclear F-actin bundles by nuclear c-Abl tyrosine kinase. (2013)
  • Author(s): Aoyama K., Yuki R., Horiike Y., Kubota S., Yamaguchi N., Morii M., Ishibashi K., Nakayama Y., Kuga T., Hashimoto Y., Tomonaga T, and Yamaguchi N.
  • Journal Title: Exp Cell Res Volume: 319 Issue: 20 Pages: 3251-68
  • DOI: 10.1016/j.yexcr.2013.09.003
  • Peer Reviewed
• [Journal Article] Phosphorylation of KRAB-associated Protein 1 (KAP1) at Tyr-449, Tyr-458, and Tyr-517 by Nuclear Tyrosine Kinases Inhibits the Association of KAP1 and Heterochromatin Protein 1alpha (HP1alpha) with Heterochromatin. (2013)
  • Author(s): Kubota, S., Fukumoto, Y., Aoyama, K., Ishibashi, K., Yuki, R., Morinaga, T., Honda, T., Yamaguchi, N., Hashimoto, Y., Kuga, T., Tomonaga T, and Yamaguchi, N.
  • Journal Title: J Biol Chem Volume: 288, Issue: 24 Pages: 17871-17883
  • DOI: 10.1074/jbc.m112.437756
  • Peer Reviewed
• [Journal Article] Serum fibrinogen alpha C-chain 5.9 kDa fragment (FIC 5.9) as a biomarker for early detection of hepatic fibrosis related to hepatitis C virus (2013)
  • Author(s): Sogawa K, Noda K, Umemura H, Seimiya M, Kuga T, Tomonaga T, Nishimura M, Kanai F, Imazeki F, Takizawa H, Yoneda M, Nakajima A, Tsutsumi M, Yokosuka O, Nomura F
  • Journal Title: Proteomics Clin Appl Volume: (in press) Issue: 5-6 Pages: 424-431
  • DOI: 10.1002/prca.201200094
  • Peer Reviewed
• [Journal Article] Identification of Missing Proteins in the neXtProt Database and Unregistered Phosphopeptides in the PhosphoSitePlus Database As Part of the Chromosome-Centric Human Proteome Project. (2013)
  • Author(s): Takashi Shiromizu
  • Journal Title: Journal of Proteome Research Volume: 印刷中 Issue: 6 Pages: 2414-2421
  • DOI: 10.1021/pr300825v
  • Peer Reviewed
• [Journal Article] A strategy for large-scale phosphoproteomics and SRM-based validation of human breast cancer tissue samples (2012)
  • Author(s): Narumi R., Murakami T., Kuga T., Adachi J., Shiromizu T., Muraoka S., Kume H., Kodera Y., Matsumoto M., Nakayama K., Miyamoto Y., Ishitobi M., Inaji H., Kato K., and Tomonaga T.
  • Journal Title: J Proteome Res Volume: 11 Pages: 5311-5322
• [Journal Article] A strategy for large-scale phosphoproteomics and SRM-based validation of human breast cancer tissue samples. (2012)
  • Author(s): Narumi R, Murakami T, Kuga T, Adachi J, Shiromizu T, Muraoka S, Kume H, Kodera Y, Matsumoto M, Nakayama K, Miyamoto Y, Ishitobi M, Inaji H, Kato K, Tomonaga T.
  • Journal Title: J Proteome Res. Volume: 11 Issue: 11 Pages: 5311-5322
  • DOI: 10.1021/pr3005474
  • Peer Reviewed
• [Presentation] FAM83H遺伝子変異に起因するエナメル質形成不全症の発症メカニズム研究 (2014)
  • Author(s): 久家貴寿,朝長毅,齊藤洋平,三上俊成,武田泰典,中山祐治
  • Organizer: 日本薬学会第134年会
  • Place of Presentation: 熊本
• [Presentation] 大腸癌細胞におけるFAM83Hとcasein kinase Iαを介したケラチン骨格制御機構の解明 (2013)
  • Author(s): 久家貴寿,久米秀明,川崎直子,足立淳,星野敢,松原久裕,齊藤洋平,中山祐治,朝長毅
  • Organizer: 第36回日本分子生物学会
  • Place of Presentation: 神戸
• [Presentation] A Novel Mechanism of Keratin Cytoskeleton Organization Through Casein Kinase I Alpha and FAM83H in Colorectal Cancer : Interactome Analysis of FAM83H (2013)
  • Author(s): Kuga T, Kume H, Kawasaki N, Sato M, Adachi J, Shiromizu T, Hoshino I, Matsubara H, Tomonaga T
  • Organizer: Human Proteome Organization (HUPO) 12^<th> Annual World Congress
  • Place of Presentation: Yokohama, Japan
• [Presentation] 大腸癌細胞におけるFAM83H と casein kinase Iαを介したケラチン骨格制御機構の解明 (2013)
  • Author(s): 久家貴寿，久米秀明，川崎直子，足立 淳，星野 敢，松原久裕，齊藤洋平，中山祐治，朝長毅
  • Organizer: 第36回日本分子生物学会
  • Place of Presentation: 神戸
• [Presentation] A Novel Mechanism of Keratin Cytoskeleton Organization Through Casein Kinase I Alpha and FAM83H in Colorectal Cancer: Interactome Analysis of FAM83H (2013)
  • Author(s): Kuga T, Kume H, Kawasaki N, Sato M, Adachi J, Shiromizu T, Hoshino I, Matsubara H, Tomonaga T
  • Organizer: Human Proteome Organization (HUPO) 12th Annual World Congress
  • Place of Presentation: 横浜
• [Presentation] annexin A2の核内蓄積はcoilin を介したセントロメア損傷によって染色体不安定性に関与する (2013)
  • Author(s): 風見隆浩，朝長 毅，川崎直子，佐藤 守，久家貴寿，松下一之，野村文夫
  • Organizer: 第36回日本分子生物学会
  • Place of Presentation: 神戸
• [Presentation] Quantitative Phosphoproteome Analysis of Cultured Stomach Cancer Cell Lines Aimed at Development of Biomarkers for Prediction of Drug Efficacy (2013)
  • Author(s): Hashiguchi K, Muraoka S, Adachi J, Sato M, Kuga T, Watanabe R, Shiromizu T, Hashimoto Y, Nagano M, Kishida M, Tomonaga T
  • Organizer: Human Proteome Organization (HUPO) 12th Annual World Congress
  • Place of Presentation: 横浜
• [Presentation] A Kinase Activity-Estimating Method Using LC-MS/MS (2013)
  • Author(s): Nagano M, Kuga T, Adachi J, Tomonaga T
  • Organizer: Human Proteome Organization (HUPO) 12th Annual World Congress
  • Place of Presentation: 横浜
• [Presentation] 新規大腸癌関連タンパク質の予後予測マーカー応用を目指した取り組み (2012)
  • Author(s): 久家貴寿
  • Organizer: 第9回千葉疾患プロテオミクス研究会
  • Place of Presentation: 東京
  • Year and Date: 2012-11-24
• [Presentation] オミックス技術を駆使した新規大腸癌関連タンパク質の同定 (2012)
  • Author(s): 久家貴寿,久米秀明,足立淳,星野敢,松原久裕,朝長毅
  • Organizer: 第71回日本癌学会学術総会
  • Place of Presentation: 札幌
• [Presentation] 大腸癌手術標本の発現解析とインタラクトーム解析による新規癌関連タンパク質の同定 (2012)
  • Author(s): 久家貴寿,久米秀明,川崎直子,足立淳,星野敢,松原久裕,朝長毅
  • Organizer: 日本プロテオーム学会2012年会
  • Place of Presentation: 東京
• [Presentation] リン酸化プロテオミクスを用いた新規DNA損傷初期応答キナーゼの探索 (2012)
  • Author(s): 足立淳
  • Organizer: 第71回日本癌学会学術総会
  • Place of Presentation: 札幌
• [Presentation] ヒト乳がん組織の大規模リン酸化プロテオーム解析とSRMをベースにした検証法 (2012)
  • Author(s): 村上達夫
  • Organizer: 第71回日本癌学会学術総会
  • Place of Presentation: 札幌
• [Presentation] リン酸化プロテオミクスを用いた新規DNA損傷初期応答キナーゼの探索 (2012)
  • Author(s): 足立淳
  • Organizer: 日本プロテオーム学会2012年会
  • Place of Presentation: 東京
• [Presentation] 大規模リン酸化プロテオーム解析とSRM/MRMによるヒト乳癌組織の検証法 (2012)
  • Author(s): 村上達夫
  • Organizer: 日本プロテオーム学会2012年会
  • Place of Presentation: 東京
• [Patent(Industrial Property Rights)] 大腸癌治療剤 (2013)
  • Inventor(s): 朝長 毅、久家貴寿、久米秀明
  • Industrial Property Rights Holder: 独立行政法人医薬基盤研究所
  • Industrial Property Rights Type: 特許
  • Filing Date: 2013-06-11
  • Overseas
• [Patent(Industrial Property Rights)] 大腸癌治療剤 (2013)
  • Inventor(s): 朝長 毅、久家貴寿、久米秀明
  • Industrial Property Rights Holder: 独立行政法人医薬基盤研究所
  • Industrial Property Rights Type: 特許
  • Filing Date: 2013-06-11
• [Patent(Industrial Property Rights)] 大腸癌治療剤 (2012)
  • Inventor(s): 朝長 毅、久家貴寿、久米秀明
  • Industrial Property Rights Holder: 独立行政法人医薬基盤研究所
  • Industrial Property Rights Type: 特許
  • Industrial Property Number: 2012-135619
  • Filing Date: 2012-06-15
• [Patent(Industrial Property Rights)] 大腸癌治療剤 (2012)
  • Inventor(s): 朝長毅、久家貴寿、久米秀明
  • Industrial Property Rights Holder: 独立行政法人医薬基盤研究所
  • Industrial Property Rights Type: 特許
  • Industrial Property Number: 2012-135619
  • Filing Date: 2012-06-15