Analysis of the urine proteome in patients with pancreatic ductal adenocarcinoma

Pancreatic ductal adenocarcinoma (PDAC) accounts for over 213 000 deaths worldwide each year, largely due to late diagnosis. One of the risk factors for the development of PDAC is chronic pancreatitis (CP); the intense desmoplastic reaction makes differentiation between the two conditions extremely difficult. In order to identify biomarkers for noninvasive diagnosis, we performed 2‐D DIGE analysis of urine samples from healthy individuals and patients with PDAC and CP. Despite considerable intersample heterogeneity, a total of 127 statistically valid (p<0.05), differentially expressed protein spots were detected, 101 of which were identified using MALDI‐TOF MS. A number of these, including annexin A2, gelsolin and CD59 have already been associated with PDAC, however, their validation using immunoblotting proved challenging. This is probably due to extensive PTMs and processing thus indicating the need for raising specific antibodies for urinary proteins. Despite this, our study clearly demonstrates that urine is a valid source of noninvasive biomarkers in patients with pancreatic diseases.

[1]  Claude Chelala,et al.  Pancreatic Expression database: a generic model for the organization, integration and mining of complex cancer datasets , 2007, BMC Genomics.

[2]  J. Barratt,et al.  Urine proteomics: the present and future of measuring urinary protein components in disease , 2007, Canadian Medical Association Journal.

[3]  Eithne Costello,et al.  The expression of S100A8 in pancreatic cancer‐associated monocytes is associated with the Smad4 status of pancreatic cancer cells , 2007, Proteomics.

[4]  K. Faber,et al.  Comparative proteomic analysis of human pancreatic juice: Methodological study , 2007, Proteomics.

[5]  P. Angel,et al.  S100A8 and S100A9 in inflammation and cancer. , 2006, Biochemical pharmacology.

[6]  Daniel F Hayes,et al.  ASCO 2006 update of recommendations for the use of tumor markers in gastrointestinal cancer. , 2006, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[7]  M. Mann,et al.  The human urinary proteome contains more than 1500 proteins, including a large proportion of membrane proteins , 2006, Genome Biology.

[8]  I. Ellis,et al.  Expression of the membrane complement regulatory protein CD59 (protectin) is associated with reduced survival in colorectal cancer patients , 2006, Cancer Immunology, Immunotherapy.

[9]  Eithne Costello,et al.  Pancreatic cancer cells overexpress gelsolin family-capping proteins, which contribute to their cell motility , 2006, Gut.

[10]  R. Aebersold,et al.  Quantitative proteomic profiling of pancreatic cancer juice , 2006, Proteomics.

[11]  John F Timms,et al.  A parallel proteomic and metabolomic analysis of the hydrogen peroxide‐ and Sty1p‐dependent stress response in Schizosaccharomyces pombe , 2006, Proteomics.

[12]  H. Friess,et al.  Tenascin C and annexin II expression in the process of pancreatic carcinogenesis , 2006, The Journal of pathology.

[13]  B. Comte,et al.  Detection of bile salt-dependent lipase, a 110 kDa pancreatic protein, in urines of healthy subjects. , 2006, Kidney international.

[14]  H. Frierson,et al.  Discovery and validation of new protein biomarkers for urothelial cancer: a prospective analysis. , 2006, The Lancet. Oncology.

[15]  R. Berkowitz,et al.  Proteomic-Based Discovery and Characterization of Glycosylated Eosinophil-Derived Neurotoxin and COOH-Terminal Osteopontin Fragments for Ovarian Cancer in Urine , 2006, Clinical Cancer Research.

[16]  Kiyoko F. Aoki-Kinoshita,et al.  From genomics to chemical genomics: new developments in KEGG , 2005, Nucleic Acids Res..

[17]  A. Olshen,et al.  Differential exoprotease activities confer tumor-specific serum peptidome patterns. , 2005, The Journal of clinical investigation.

[18]  Jue Wang,et al.  Human urine proteome analysis by three separation approaches , 2005, Proteomics.

[19]  Kenoki Ohuchida,et al.  The Role of S100A6 in Pancreatic Cancer Development and Its Clinical Implication as a Diagnostic Marker and Therapeutic Target , 2005, Clinical Cancer Research.

[20]  Fiona Campbell,et al.  Proteomic analysis of chronic pancreatitis and pancreatic adenocarcinoma. , 2005, Gastroenterology.

[21]  Ruedi Aebersold,et al.  Pancreatic cancer proteome: the proteins that underlie invasion, metastasis, and immunologic escape. , 2005, Gastroenterology.

[22]  A. Rustgi,et al.  Characterization of proteins in human pancreatic cancer serum using differential gel electrophoresis and tandem mass spectrometry. , 2005, Journal of proteome research.

[23]  M. Waterfield,et al.  Proteomic analysis of redox‐ and ErbB2‐dependent changes in mammary luminal epithelial cells using cysteine‐ and lysine‐labelling two‐dimensional difference gel electrophoresis , 2005, Proteomics.

[24]  G. Klöppel,et al.  Application of fluorescence difference gel electrophoresis saturation labelling for the analysis of microdissected precursor lesions of pancreatic ductal adenocarcinoma , 2005, Proteomics.

[25]  R. Tonge,et al.  Development of a high‐throughput method for preparing human urine for two‐dimensional electrophoresis , 2005, Proteomics.

[26]  E. Nexo,et al.  Glycosylation independent measurement of the cobalamin binding protein haptocorrin. , 2005, Clinica chimica acta; international journal of clinical chemistry.

[27]  Shui-Tein Chen,et al.  Different techniques for urinary protein analysis of normal and lung cancer patients , 2005, Proteomics.

[28]  V. Thongboonkerd Genomics, proteomics and integrative ‘omics’ in hypertension research , 2005, Current opinion in nephrology and hypertension.

[29]  Kevin R Coombes,et al.  Plasma protein profiling for diagnosis of pancreatic cancer reveals the presence of host response proteins. , 2005, Clinical cancer research : an official journal of the American Association for Cancer Research.

[30]  James L. Abbruzzese,et al.  Protein Expression Profiles in Pancreatic Adenocarcinoma Compared with Normal Pancreatic Tissue and Tissue Affected by Pancreatitis as Detected by Two-Dimensional Gel Electrophoresis and Mass Spectrometry , 2004, Cancer Research.

[31]  D. Zurakowski,et al.  ADAM 12 Cleaves Extracellular Matrix Proteins and Correlates with Cancer Status and Stage* , 2004, Journal of Biological Chemistry.

[32]  Zhaohui Lu,et al.  Differential expression profiling of human pancreatic adenocarcinoma and healthy pancreatic tissue , 2004, Proteomics.

[33]  N. Stergiopulos,et al.  Gelsolin superfamily proteins: key regulators of cellular functions , 2004, Cellular and Molecular Life Sciences CMLS.

[34]  Troels Z. Kristiansen,et al.  Comprehensive proteomic analysis of human pancreatic juice. , 2004, Journal of proteome research.

[35]  E. Birney,et al.  The International Protein Index: An integrated database for proteomics experiments , 2004, Proteomics.

[36]  F. Motoi,et al.  Clinicopathological Aspects of Small Pancreatic Cancer , 2004, Pancreas.

[37]  Rembert Pieper,et al.  Characterization of the human urinary proteome: A method for high‐resolution display of urinary proteins on two‐dimensional electrophoresis gels with a yield of nearly 1400 distinct protein spots , 2004, Proteomics.

[38]  H. Ploegh,et al.  Zinc-finger protein A20, a regulator of inflammation and cell survival, has de-ubiquitinating activity. , 2004, The Biochemical journal.

[39]  Frank Schmidt,et al.  Iterative data analysis is the key for exhaustive analysis of peptide mass fingerprints from proteins separated by two-dimensional electrophoresis , 2003, Journal of the American Society for Mass Spectrometry.

[40]  S. Pinder,et al.  Loss of CD59 expression in breast tumours correlates with poor survival , 2003, The Journal of pathology.

[41]  Tatjana Crnogorac-Jurcevic,et al.  Expression profiling of microdissected pancreatic adenocarcinomas , 2002, Oncogene.

[42]  C. Rosty,et al.  Early detection of pancreatic carcinoma. , 2002, Hematology/oncology clinics of North America.

[43]  R. Wahl,et al.  Towards defining the urinary proteome using liquid chromatography‐tandem mass spectrometry I.Profiling an unfractionated tryptic digest , 2001, Proteomics.

[44]  D. Winchester,et al.  Pancreatic cancer: a report of treatment and survival trends for 100,313 patients diagnosed from 1985-1995, using the National Cancer Database. , 1999, Journal of the American College of Surgeons.

[45]  P. Sims,et al.  Elimination of Potential Sites of Glycosylation Fails to Abrogate Complement Regulatory Function of Cell Surface CD59* , 1996, The Journal of Biological Chemistry.

[46]  D. Waisman Annexin II tetramer: structure and function , 1995, Molecular and Cellular Biochemistry.

[47]  E. Ulvestad,et al.  Identification of a soluble Fcy‐binding molecule (annexin II) in huan serum using a competitive ELISA , 1994 .

[48]  N. Anderson,et al.  Proteins of human urine. I. Concentration and analysis by two-dimensional electrophoresis. , 1979, Clinical chemistry.

[49]  Charles Darwin,et al.  Experiments , 1800, The Medical and physical journal.

[50]  A. Jemal,et al.  Cancer Statistics, 2007 , 2007, CA: a cancer journal for clinicians.

[51]  K. Ohuchida,et al.  The Role of S 100 A 6 in Pancreatic Cancer Development and Its Clinical Implication as a DiagnosticMarker and Therapeutic Target , 2005 .

[52]  P. Nickerson,et al.  Urine protein profiling with surface-enhanced laser-desorption/ionization time-of-flight mass spectrometry. , 2004, Kidney international.

[53]  P. V. Bottini,et al.  Electrophoretic pattern of concentrated urine: comparison between 24-hour collection and random samples. , 2002, American journal of kidney diseases : the official journal of the National Kidney Foundation.

[54]  E. R. Taylor,et al.  Isolation and characterization of two novel A20-like proteins. , 2001, The Biochemical journal.

[55]  E. Ulvestad,et al.  Identification of a soluble Fc gamma-binding molecule (annexin II) in human serum using a competitive ELISA. , 1994, APMIS : acta pathologica, microbiologica, et immunologica Scandinavica.

[56]  N. Dubrawsky Cancer statistics , 2022 .