Identification of potential serum peptide biomarkers of biliary tract cancer using MALDI MS profiling

BMC Clinical Pathology

Stephen P Pereira¹  John F Timms²  Ross C Smith³  George J Webster¹  Michael H Chapman¹  Fausto Andreola⁴  Oleg Blyuss²  John Sinclair²  Stephane Camuzeaux²  Neomal S Sandanayake³ 

[1] Department of Gastroenterology, University College Hospitals NHS Foundation Trust, London, UK;Cancer Proteomics Laboratory, EGA Institute for Women’s Health, University College London, London, UK;Kolling Institute, University of Sydney, St Leonards, Australia;UCL Institute for Liver and Digestive Health, Royal Free Hospital Campus, University College London, London, UK

关键词: MALDI-TOF MS;    Serum peptide biomarker;    PSC;    Biliary tract cancer;    Cholangiocarcinoma;   

Others  :  819998
DOI  :  10.1186/1472-6890-14-7

 received in 2013-07-13, accepted in 2014-01-13,  发布年份 2014

【 摘 要 】

Background

The aim of this discovery study was the identification of peptide serum biomarkers for detecting biliary tract cancer (BTC) using samples from healthy volunteers and benign cases of biliary disease as control groups. This work was based on the hypothesis that cancer-specific exopeptidases exist and that their activities in serum can generate cancer-predictive peptide fragments from circulating proteins during coagulation.

Methods

This case control study used a semi-automated platform incorporating polypeptide extraction linked to matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry (MALDI-TOF MS) to profile 92 patient serum samples. Predictive models were generated to test a validation serum set from BTC cases and healthy volunteers.

Results

Several peptide peaks were found that could significantly differentiate BTC patients from healthy controls and benign biliary disease. A predictive model resulted in a sensitivity of 100% and a specificity of 93.8% in detecting BTC in the validation set, whilst another model gave a sensitivity of 79.5% and a specificity of 83.9% in discriminating BTC from benign biliary disease samples in the training set. Discriminatory peaks were identified by tandem MS as fragments of abundant clotting proteins.

Conclusions

Serum MALDI MS peptide signatures can accurately discriminate patients with BTC from healthy volunteers.

【 授权许可】

   
2014 Sandanayake et al.; licensee BioMed Central Ltd.

【 图 表 】

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【 参考文献 】

• [1]Rizvi S, Gores GJ: Pathogenesis, diagnosis, and management of cholangiocarcinoma. Gastroenterology 2013, 145(6):1215-29.
• [2]Khan SA, Taylor-Robinson SD, Toledano MB, Beck A, Elliott P, Thomas HC: Changing international trends in mortality rates for liver, biliary and pancreatic tumours. J Hepatol 2002, 37(6):806-813.
• [3]Patel T: Increasing incidence and mortality of primary intrahepatic cholangiocarcinoma in the United States. Hepatology 2001, 33(6):1353-1357.
• [4]Jarnagin WR, Fong Y, DeMatteo RP, Gonen M, Burke EC, Bodniewicz BJ, Youssef BM, Klimstra D, Blumgart LH: Staging, resectability, and outcome in 225 patients with hilar cholangiocarcinoma. Ann Surg 2001, 234(4):507-517. discussion 517–509
• [5]Nakagawa T, Kamiyama T, Kurauchi N, Matsushita M, Nakanishi K, Kamachi H, Kudo T, Todo S: Number of lymph node metastases is a significant prognostic factor in intrahepatic cholangiocarcinoma. World J Surg 2005, 29(6):728-733.
• [6]Ohtsuka M, Ito H, Kimura F, Shimizu H, Togawa A, Yoshidome H, Miyazaki M: Results of surgical treatment for intrahepatic cholangiocarcinoma and clinicopathological factors influencing survival. Br J Surg 2002, 89(12):1525-1531.
• [7]Skipworth JRA, Timms JF, Pereira SP: Novel diagnostic and prognostic biomarkers in biliary tract cancer. Expert Opin Med Diagn 2013, 7(5):487-99.
• [8]Lamerz R: Role of tumour markers, cytogenetics. Ann Oncol 1999, 10(Suppl 4):145-149.
• [9]Steinberg W: The clinical utility of the CA 19–9 tumor-associated antigen. Am J Gastroenterol 1990, 85(4):350-355.
• [10]Ebert MP, Meuer J, Wiemer JC, Schulz HU, Reymond MA, Traugott U, Malfertheiner P, Rocken C: Identification of gastric cancer patients by serum protein profiling. J Proteome Res 2004, 3(6):1261-1266.
• [11]Hingorani SR, Petricoin EF, Maitra A, Rajapakse V, King C, Jacobetz MA, Ross S, Conrads TP, Veenstra TD, Hitt BA, et al.: Preinvasive and invasive ductal pancreatic cancer and its early detection in the mouse. Cancer Cell 2003, 4(6):437-450.
• [12]Villanueva J, Philip J, Entenberg D, Chaparro CA, Tanwar MK, Holland EC, Tempst P: Serum peptide profiling by magnetic particle-assisted, automated sample processing and MALDI-TOF mass spectrometry. Anal Chem 2004, 76(6):1560-1570.
• [13]Villanueva J, Shaffer DR, Philip J, Chaparro CA, Erdjument-Bromage H, Olshen AB, Fleisher M, Lilja H, Brogi E, Boyd J, et al.: Differential exoprotease activities confer tumor-specific serum peptidome patterns. J Clin Invest 2006, 116(1):271-284.
• [14]Sobin LH, Fleming ID: TNM classification of malignant tumors, fifth edition (1997). Union Internationale Contre le Cancer and the American Joint Committee on Cancer. Cancer 1997, 80(9):1803-1804.
• [15]Timms JF, Cramer R, Camuzeaux S, Tiss A, Smith C, Burford B, Nouretdinov I, Devetyarov D, Gentry-Maharaj A, Ford J, et al.: Peptides generated ex vivo from serum proteins by tumor-specific exopeptidases are not useful biomarkers in ovarian cancer. Clin Chem 2010, 56(2):262-271.
• [16]Koomen JM, Li D, Xiao LC, Liu TC, Coombes KR, Abbruzzese J, Kobayashi R: Direct tandem mass spectrometry reveals limitations in protein profiling experiments for plasma biomarker discovery. J Proteome Res 2005, 4(3):972-981.
• [17]Tiss A, Smith C, Menon U, Jacobs I, Timms JF, Cramer R: A well-characterised peak identification list of MALDI MS profile peaks for human blood serum. Proteomics 2010, 10(18):3388-3392.
• [18]Petricoin EF, Ardekani AM, Hitt BA, Levine PJ, Fusaro VA, Steinberg SM, Mills GB, Simone C, Fishman DA, Kohn EC, et al.: Use of proteomic patterns in serum to identify ovarian cancer. Lancet 2002, 359(9306):572-577.
• [19]Aebersold R, Mann M: Mass spectrometry-based proteomics. Nature 2003, 422(6928):198-207.
• [20]Liotta LA, Ferrari M, Petricoin E: Clinical proteomics: written in blood. Nature 2003, 425(6961):905.
• [21]Rai AJ, Chan DW: Cancer proteomics: serum diagnostics for tumor marker discovery. Ann N Y Acad Sci 2004, 1022:286-294.
• [22]Coombes KR, Morris JS, Hu J, Edmonson SR, Baggerly KA: Serum proteomics profiling–a young technology begins to mature. Nat Biotechnol 2005, 23(3):291-292.
• [23]Diamandis EP: Analysis of serum proteomic patterns for early cancer diagnosis: drawing attention to potential problems. J Natl Cancer Inst 2004, 96(5):353-356.
• [24]Baggerly KA, Morris JS, Coombes KR: Reproducibility of SELDI-TOF protein patterns in serum: comparing datasets from different experiments. Bioinformatics 2004, 20(5):777-785.
• [25]Ransohoff DF: Lessons from controversy: ovarian cancer screening and serum proteomics. J Natl Cancer Inst 2005, 97(4):315-319.
• [26]Diamandis EP: Mass spectrometry as a diagnostic and a cancer biomarker discovery tool: opportunities and potential limitations. Mol Cell Proteomics 2004, 3(4):367-378.
• [27]Hortin GL: Can mass spectrometric protein profiling meet desired standards of clinical laboratory practice? Clin Chem 2005, 51(1):3-5.
• [28]Villanueva J, Lawlor K, Toledo-Crow R, Tempst P: Automated serum peptide profiling. Nat Protoc 2006, 1(2):880-891.
• [29]Villanueva J, Martorella AJ, Lawlor K, Philip J, Fleisher M, Robbins RJ, Tempst P: Serum peptidome patterns that distinguish metastatic thyroid carcinoma from cancer-free controls are unbiased by gender and age. Mol Cell Proteomics 2006, 5(10):1840-1852.
• [30]Lewis JT, Talwalkar JA, Rosen CB, Smyrk TC, Abraham SC: Precancerous bile duct pathology in end-stage primary sclerosing cholangitis, with and without cholangiocarcinoma. Am J Surg Pathol 2010, 34(1):27-34.
• [31]Fiedler GM, Leichtle AB, Kase J, Baumann S, Ceglarek U, Felix K, Conrad T, Witzigmann H, Weimann A, Schutte C, et al.: Serum peptidome profiling revealed platelet factor 4 as a potential discriminating Peptide associated with pancreatic cancer. Clin Cancer Res 2009, 15(11):3812-3819.
• [32]Zabron AA, der Sluis VM H-v, Wadsworth CA, Laird F, Gierula M, Thillainayagam AV, Vlavianos P, Westaby D, Taylor-Robinson SD, Edwards RJ, et al.: Elevated levels of neutrophil gelatinase-associated lipocalin in bile from patients with malignant pancreatobiliary disease. Am J Gastroenterol 2011, 106(9):1711-1717.
• [33]Mitsui T, Nomura S, Itakura A, Mizutani S: Role of aminopeptidases in the blood pressure regulation. Biol Pharm Bull 2004, 27(6):768-771.
• [34]Nesheim M, Wang W, Boffa M, Nagashima M, Morser J, Bajzar L: Thrombin, thrombomodulin and TAFI in the molecular link between coagulation and fibrinolysis. Thromb Haemost 1997, 78(1):386-391.
• [35]van Hensbergen Y, Broxterman HJ, Hanemaaijer R, Jorna AS, van Lent NA, Verheul HM, Pinedo HM, Hoekman K: Soluble aminopeptidase N/CD13 in malignant and nonmalignant effusions and intratumoral fluid. Clin Cancer Res 2002, 8(12):3747-3754.
• [36]Moffatt S, Wiehle S, Cristiano RJ: Tumor-specific gene delivery mediated by a novel peptide-polyethylenimine-DNA polyplex targeting aminopeptidase N/CD13. Hum Gene Ther 2005, 16(1):57-67.
• [37]Kehlen A, Lendeckel U, Dralle H, Langner J, Hoang-Vu C: Biological significance of aminopeptidase N/CD13 in thyroid carcinomas. Cancer Res 2003, 63(23):8500-8506.
• [38]Rocken C, Carl-McGrath S, Grantzdorffer I, Mantke R, Roessner A, Lendeckel U: Ectopeptidases are differentially expressed in hepatocellular carcinomas. Int J Oncol 2004, 24(3):487-495.
• [39]Pasqualini R, Koivunen E, Kain R, Lahdenranta J, Sakamoto M, Stryhn A, Ashmun RA, Shapiro LH, Arap W, Ruoslahti E: Aminopeptidase N is a receptor for tumor-homing peptides and a target for inhibiting angiogenesis. Cancer Res 2000, 60(3):722-727.
• [40]Theodorescu D, Wittke S, Ross MM, Walden M, Conaway M, Just I, Mischak H, Frierson HF: Discovery and validation of new protein biomarkers for urothelial cancer: a prospective analysis. Lancet Oncol 2006, 7(3):230-240.
• [41]Bergen HR 3rd, Vasmatzis G, Cliby WA, Johnson KL, Oberg AL, Muddiman DC: Discovery of ovarian cancer biomarkers in serum using NanoLC electrospray ionization TOF and FT-ICR mass spectrometry. Dis Markers 2003, 19(4–5):239-249.
• [42]Orvisky E, Drake SK, Martin BM, Abdel-Hamid M, Ressom HW, Varghese RS, An Y, Saha D, Hortin GL, Loffredo CA, et al.: Enrichment of low molecular weight fraction of serum for MS analysis of peptides associated with hepatocellular carcinoma. Proteomics 2006, 6(9):2895-2902.
• [43]Ebert MP, Niemeyer D, Deininger SO, Wex T, Knippig C, Hoffmann J, Sauer J, Albrecht W, Malfertheiner P, Rocken C: Identification and confirmation of increased fibrinopeptide a serum protein levels in gastric cancer sera by magnet bead assisted MALDI-TOF mass spectrometry. J Proteome Res 2006, 5(9):2152-2158.