Identification of IGFBP2 and IGFBP3 As Compensatory Biomarkers for CA19-9 in Early-Stage Pancreatic Cancer Using a Combination of Antibody-Based and LC-MS/MS-Based Proteomics

Toshihiro Yoneyama; Sumio Ohtsuki; Kazufumi Honda; Makoto Kobayashi; Motoki Iwasaki; Yasuo Uchida; Takuji Okusaka; Shoji Nakamori; Masashi Shimahara; Takaaki Ueno; Akihiko Tsuchida; Naohiro Sata; Tatsuya Ioka; Yohichi Yasunami; Tomoo Kosuge; Takashi Kaneda; Takao Kato; Kazuhiro Yagihara; Shigeyuki Fujita; Wilber Huang; Tesshi Yamada; Masanori Tachikawa; Tetsuya Terasaki

doi:10.1371/journal.pone.0161009

Identification of IGFBP2 and IGFBP3 As Compensatory Biomarkers for CA19-9 in Early-Stage Pancreatic Cancer Using a Combination of Antibody-Based and LC-MS/MS-Based Proteomics

PLoS One. 2016 Aug 31;11(8):e0161009. doi: 10.1371/journal.pone.0161009. eCollection 2016.

Authors

Toshihiro Yoneyama¹, Sumio Ohtsuki^{2

3}, Kazufumi Honda^{4

3}, Makoto Kobayashi⁴, Motoki Iwasaki⁵, Yasuo Uchida¹, Takuji Okusaka⁶, Shoji Nakamori⁷, Masashi Shimahara⁸, Takaaki Ueno⁸, Akihiko Tsuchida⁹, Naohiro Sata¹⁰, Tatsuya Ioka¹¹, Yohichi Yasunami¹², Tomoo Kosuge¹³, Takashi Kaneda¹⁴, Takao Kato¹⁵, Kazuhiro Yagihara¹⁶, Shigeyuki Fujita¹⁷, Wilber Huang¹⁸, Tesshi Yamada⁴, Masanori Tachikawa¹, Tetsuya Terasaki¹

Affiliations

¹ Division of Membrane Transport and Drug Targeting, Graduate School of Pharmaceutical Sciences, Tohoku University, Sendai, Japan.
² Department of Pharmaceutical Microbiology, Faculty of Life Sciences, Kumamoto University, Kumamoto, Japan.
³ Japan Agency for Medical Research and Development (AMED) CREST, Tokyo, Japan.
⁴ Division of Chemotherapy and Clinical Research, National Cancer Center Research Institute, Tokyo, Japan.
⁵ Division of Epidemiology, Research Center for Cancer Prevention and Screening, National Cancer Center, Tokyo, Japan.
⁶ Department of Hepatobiliary and Pancreatic Oncology, National Cancer Center Hospital, Tokyo, Japan.
⁷ Departments of Hepato-Biliary-Pancreatic Surgery, Osaka National Hospital, National Hospital Organization, Osaka, Japan.
⁸ Department of Oral Surgery, Osaka Medical College, Osaka, Japan.
⁹ Department of Gastrointestinal and Pediatric Surgery, Tokyo Medical University, Tokyo, Japan.
¹⁰ Department of Surgery, Jichi Medical University, Tochigi, Japan.
¹¹ Department of Hepatobiliary and Pancreatic Oncology, Osaka Medical Center for Cancer and Cardiovascular Diseases, Osaka, Japan.
¹² Islet Institute, Fukuoka University, Fukuoka, Japan.
¹³ Hepatobiliary and Pancreatic Surgery Division, National Cancer Center Hospital, Tokyo, Japan.
¹⁴ Department of Radiology, Nihon University School of Dentistry at Matsudo, Chiba, Japan.
¹⁵ Department of Oral Implant, Nihon University School of Dentistry at Matsudo, Chiba, Japan.
¹⁶ Department of Oral Surgery, Saitama Cancer Center, Saitama, Japan.
¹⁷ Department of Oral and Maxillofacial Surgery, Wakayama Medical University, Wakayama, Japan.
¹⁸ Abnova, Taipei City, Taiwan.

Abstract

Pancreatic cancer is one of the most lethal tumors, and reliable detection of early-stage pancreatic cancer and risk diseases for pancreatic cancer is essential to improve the prognosis. As 260 genes were previously reported to be upregulated in invasive ductal adenocarcinoma of pancreas (IDACP) cells, quantification of the corresponding proteins in plasma might be useful for IDACP diagnosis. Therefore, the purpose of the present study was to identify plasma biomarkers for early detection of IDACP by using two proteomics strategies: antibody-based proteomics and liquid chromatography-tandem mass spectrometry (LC-MS/MS)-based proteomics. Among the 260 genes, we focused on 130 encoded proteins with known function for which antibodies were available. Twenty-three proteins showed values of the area under the curve (AUC) of more than 0.8 in receiver operating characteristic (ROC) analysis of reverse-phase protein array (RPPA) data of IDACP patients compared with healthy controls, and these proteins were selected as biomarker candidates. We then used our high-throughput selected reaction monitoring or multiple reaction monitoring (SRM/MRM) methodology, together with an automated sample preparation system, micro LC and auto analysis system, to quantify these candidate proteins in plasma from healthy controls and IDACP patients on a large scale. The results revealed that insulin-like growth factor-binding protein (IGFBP)2 and IGFBP3 have the ability to discriminate IDACP patients at an early stage from healthy controls, and IGFBP2 appeared to be increased in risk diseases of pancreatic malignancy, such as intraductal papillary mucinous neoplasms (IPMNs). Furthermore, diagnosis of IDACP using the combination of carbohydrate antigen 19-9 (CA19-9), IGFBP2 and IGFBP3 is significantly more effective than CA19-9 alone. This suggests that IGFBP2 and IGFBP3 may serve as compensatory biomarkers for CA19-9. Early diagnosis with this marker combination may improve the prognosis of IDACP patients.

Publication types

Clinical Trial
Multicenter Study

MeSH terms

Adult
Aged
Biomarkers, Tumor / blood*
CA-19-9 Antigen / blood*
Carcinoma, Pancreatic Ductal / blood*
Carcinoma, Pancreatic Ductal / diagnosis
Chromatography, Liquid
Female
Humans
Insulin-Like Growth Factor Binding Protein 2 / blood*
Insulin-Like Growth Factor Binding Protein 3 / blood*
Male
Mass Spectrometry
Middle Aged
Neoplasm Proteins / blood*
Neoplasm Staging
Pancreatic Neoplasms / blood*
Pancreatic Neoplasms / diagnosis
Pancreatic Neoplasms / pathology
Prognosis
Proteomics*

Substances

Biomarkers, Tumor
CA-19-9 Antigen
IGFBP2 protein, human
IGFBP3 protein, human
Insulin-Like Growth Factor Binding Protein 2
Insulin-Like Growth Factor Binding Protein 3
Neoplasm Proteins

Grants and funding

This study was performed as a research program in the Project for Development of Innovative Research on Cancer Therapeutics (P-Direct, 14089014), Ministry of Education, Culture, Sports, Science and Technology of Japan (http://www.mext.go.jp). This study was also supported in part by JSPS KAKENHI Grant Numbers 14J05219, 15K15751, 23390469, and by the Industrial Technology Research Grant Program of the New Energy and the Industrial Technology Development Organization of Japan (P00041), and the Funding Program for Next Generation World-Leading Researchers (LS005) by the Cabinet Office, Government of Japan (http://www.cao.go.jp/index-e.html), and the Third-Term Comprehensive Control Research for Cancer, Research on Biological Markers for New Drug Development and Health Labour Sciences Research, and Applied Research for Innovative Treatment of Cancer grant by the Ministry of Health, Labour and Welfare of Japan, and Practical Research for Innovation Cancer Control (15ck0106101h0002), and AMED-CREST (10801082) by Japan Agency for Medical Research and Development (http://www.amed.go.jp/). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. One of our authors, W.H., is a CEO of a commercial company: Abnova. The funder provided support in the form of salaries for W.H. and research materials (antibodies for RPPA), but did not have any additional role in the study design, data collection and analysis, decision to publish, or preparation of the manuscript.