Protein and glycomic plasma markers for early detection of adenoma and colon cancer

Objective To discover and confirm blood-based colon cancer early-detection markers. Design We created a high-density antibody microarray to detect differences in protein levels in plasma from individuals diagnosed with colon cancer <3 years after blood was drawn (ie, prediagnostic) and cancer-free, matched controls. Potential markers were tested on plasma samples from people diagnosed with adenoma or cancer, compared with controls. Components of an optimal 5-marker panel were tested via immunoblotting using a third sample set, Luminex assay in a large fourth sample set and immunohistochemistry (IHC) on tissue microarrays. Results In the prediagnostic samples, we found 78 significantly (t-test) increased proteins, 32 of which were confirmed in the diagnostic samples. From these 32, optimal 4-marker panels of BAG family molecular chaperone regulator 4 (BAG4), interleukin-6 receptor subunit beta (IL6ST), von Willebrand factor (VWF) and CD44 or epidermal growth factor receptor (EGFR) were established. Each panel member and the panels also showed increases in the diagnostic adenoma and cancer samples in independent third and fourth sample sets via immunoblot and Luminex, respectively. IHC results showed increased levels of BAG4, IL6ST and CD44 in adenoma and cancer tissues. Inclusion of EGFR and CD44 sialyl Lewis-A and Lewis-X content increased the panel performance. The protein/glycoprotein panel was statistically significantly higher in colon cancer samples, characterised by a range of area under the curves from 0.90 (95% CI 0.82 to 0.98) to 0.86 (95% CI 0.83 to 0.88), for the larger second and fourth sets, respectively. Conclusions A panel including BAG4, IL6ST, VWF, EGFR and CD44 protein/glycomics performed well for detection of early stages of colon cancer and should be further examined in larger studies.

[1]  M. Phipps,et al.  Screening for Colorectal Cancer: US Preventive Services Task Force Recommendation Statement. , 2016, JAMA.

[2]  Yuzheng Zhang,et al.  Cross-Species Antibody Microarray Interrogation Identifies a 3-Protein Panel of Plasma Biomarkers for Early Diagnosis of Pancreas Cancer , 2015, Clinical Cancer Research.

[3]  S. Winawer Long-term colorectal-cancer mortality after adenoma removal. , 2014, The New England journal of medicine.

[4]  M. Hollingsworth,et al.  Spatiotemporal Proteomic Analyses during Pancreas Cancer Progression Identifies Serine/Threonine Stress Kinase 4 (STK4) as a Novel Candidate Biomarker for Early Stage Disease* , 2014, Molecular & Cellular Proteomics.

[5]  D. Ransohoff,et al.  Multitarget stool DNA testing for colorectal-cancer screening. , 2014, The New England journal of medicine.

[6]  J. Stave,et al.  Discovery of sialyl Lewis A and Lewis X modified protein cancer biomarkers using high density antibody arrays. , 2014, Journal of proteomics.

[7]  Reiko Nishihara,et al.  Long-term colorectal-cancer incidence and mortality after lower endoscopy. , 2013, The New England journal of medicine.

[8]  P. Lampe,et al.  High-throughput screening for native autoantigen-autoantibody complexes using antibody microarrays. , 2013, Journal of proteome research.

[9]  Thomas Rösch,et al.  Prospective evaluation of methylated SEPT9 in plasma for detection of asymptomatic colorectal cancer , 2013, Gut.

[10]  S. Hanash,et al.  Discovery and preliminary confirmation of novel early detection biomarkers for triple-negative breast cancer using preclinical plasma samples from the Women’s Health Initiative observational study , 2012, Breast Cancer Research and Treatment.

[11]  S. Thibodeau,et al.  The stool DNA test is more accurate than the plasma septin 9 test in detecting colorectal neoplasia. , 2012, Clinical gastroenterology and hepatology : the official clinical practice journal of the American Gastroenterological Association.

[12]  S. Hanash,et al.  Increased Plasma Levels of the APC-Interacting Protein MAPRE1, LRG1, and IGFBP2 Preceding a Diagnosis of Colorectal Cancer in Women , 2012, Cancer Prevention Research.

[13]  Chien-Yu Chen,et al.  Sialylation and fucosylation of epidermal growth factor receptor suppress its dimerization and activation in lung cancer cells , 2011, Proceedings of the National Academy of Sciences.

[14]  N. Urban,et al.  Use of a Single-Chain Antibody Library for Ovarian Cancer Biomarker Discovery* , 2010, Molecular & Cellular Proteomics.

[15]  R. Kopp,et al.  Frequent expression of the high molecular, 673-bp CD44v3,v8-10 variant in colorectal adenomas and carcinomas. , 2009, International journal of molecular medicine.

[16]  Richard J. Simpson,et al.  Proteomics Analysis of A33 Immunoaffinity-purified Exosomes Released from the Human Colon Tumor Cell Line LIM1215 Reveals a Tissue-specific Protein Signature* , 2009, Molecular & Cellular Proteomics.

[17]  Holly Janes,et al.  Pivotal Evaluation of the Accuracy of a Biomarker Used for Classification or Prediction: Standards for Study Design , 2008, Journal of the National Cancer Institute.

[18]  R. Kannagi,et al.  Clinical application of functional glycoproteomics – dissection of glycotopes carried by soluble CD44 variants in sera of patients with cancers , 2008, Proteomics.

[19]  Xin-Yun Huang,et al.  Identification of Novel Direct Stat3 Target Genes for Control of Growth and Differentiation* , 2008, Journal of Biological Chemistry.

[20]  Philip R. Gafken,et al.  Phosphorylation at S365 is a gatekeeper event that changes the structure of Cx43 and prevents down-regulation by PKC , 2007, The Journal of cell biology.

[21]  J. Delrow,et al.  Use of high density antibody arrays to validate and discover cancer serum biomarkers , 2007, Molecular oncology.

[22]  M Westwood,et al.  Diagnostic accuracy of faecal occult blood tests used in screening for colorectal cancer: a systematic review , 2007, Journal of medical screening.

[23]  G. Stark,et al.  Unphosphorylated STAT3 accumulates in response to IL-6 and activates transcription by binding to NFkappaB. , 2007, Genes & development.

[24]  Dean Brenner,et al.  Multiplexed analysis of glycan variation on native proteins captured by antibody microarrays , 2007, Nature Methods.

[25]  A. Siriwardena,et al.  Systematic review of carbohydrate antigen (CA 19-9) as a biochemical marker in the diagnosis of pancreatic cancer. , 2007, European journal of surgical oncology : the journal of the European Society of Surgical Oncology and the British Association of Surgical Oncology.

[26]  J. Davis Bioinformatics and Computational Biology Solutions Using R and Bioconductor , 2007 .

[27]  Alicia Oshlack,et al.  Normalization of boutique two-color microarrays with a high proportion of differentially expressed probes , 2007, Genome Biology.

[28]  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.

[29]  Nancy Breen,et al.  Patterns of Colorectal Cancer Screening Uptake among Men and Women in the United States , 2006, Cancer Epidemiology Biomarkers & Prevention.

[30]  K. Kinzler,et al.  Somatic mutations of EGFR in colorectal cancers and glioblastomas. , 2004, The New England journal of medicine.

[31]  L. Seeff,et al.  Is there endoscopic capacity to provide colorectal cancer screening to the unscreened population in the United States? , 2004, Gastroenterology.

[32]  M. Lakshman,et al.  CD44 promotes resistance to apoptosis in human colon cancer cells. , 2004, Experimental and molecular pathology.

[33]  Terry Speed,et al.  Normalization of cDNA microarray data. , 2003, Methods.

[34]  J. Potter,et al.  Long-term efficacy of sigmoidoscopy in the reduction of colorectal cancer incidence. , 2003, Journal of the National Cancer Institute.

[35]  John D. Storey A direct approach to false discovery rates , 2002 .

[36]  John Calvin Reed,et al.  Molecular chaperone targeting and regulation by BAG family proteins , 2001, Nature Cell Biology.

[37]  M S Pepe,et al.  Phases of biomarker development for early detection of cancer. , 2001, Journal of the National Cancer Institute.

[38]  M. Duffy,et al.  Carcinoembryonic antigen as a marker for colorectal cancer: is it clinically useful? , 2001, Clinical chemistry.

[39]  P. Brown,et al.  Protein microarrays for highly parallel detection and quantitation of specific proteins and antibodies in complex solutions , 2001, Genome Biology.

[40]  D. Allred,et al.  Prognostic and predictive factors in breast cancer by immunohistochemical analysis. , 1998, Modern pathology : an official journal of the United States and Canadian Academy of Pathology, Inc.

[41]  J. Cuzick,et al.  Prevention of colorectal cancer by once-only sigmoidoscopy , 1993, The Lancet.

[42]  J Cuzick,et al.  Long-term risk of colorectal cancer after excision of rectosigmoid adenomas. , 1992, The New England journal of medicine.

[43]  R. Kronmal,et al.  The Cardiovascular Health Study: design and rationale. , 1991, Annals of epidemiology.

[44]  Hye-Jeong Song,et al.  Exploring Multiple Biomarker Combination by Logistic Regression for Early Screening of Ovarian Cancer , 2013 .

[45]  Ruth M Pfeiffer,et al.  Proteomic biomarkers in combination with CA 125 for detection of epithelial ovarian cancer using prediagnostic serum samples from the Prostate, Lung, Colorectal, and Ovarian (PLCO) Cancer Screening Trial , 2012, Cancer.

[46]  Gordon K. Smyth,et al.  limma: Linear Models for Microarray Data , 2005 .

[47]  T. Ørntoft,et al.  Reference values and biological variation for tumor marker CA 19-9 in serum for different Lewis and secretor genotypes and evaluation of secretor and Lewis genotyping in a Caucasian population. , 1999, Clinical chemistry.

[48]  F. V. von Eyben,et al.  Colorectal cancer screening: clinical guidelines and rationale. , 1997, Gastroenterology.

[49]  Robert J. Mayer,et al.  Erratum: Colorectal cancer screening: Clinical guidelines and rationale (Gastroenterology (1997) 112 (594-642)) , 1997 .