title: Plasma clusterin as a candidate pre-diagnosis marker of colorectal cancer risk in the Florence cohort of the European Prospective Investigation into Cancer and Nutrition: a pilot study.
abstract: BACKGROUND
  Colorectal cancer is one of the major causes of cancer mortality world-wide. Prevention would improve if at-risk subjects could be identified. The aim of this study was to characterise plasma protein biomarkers associated with the risk of colorectal cancer in samples collected prospectively, before the disease diagnosis.
  
  
  METHODS
  After an exploratory study on the comprehensive plasma proteome analysis by liquid chromatography-tandem mass spectrometry from ten colorectal cancer cases enrolled at diagnosis, and ten matched controls (Phase 1), a similar preliminary study was performed on prospective plasma samples from ten colorectal cancer cases, enrolled years before disease development, and ten matched controls identified in a nested case-control study within the Florence cohort of the European Prospective Investigation into Cancer and Nutrition (EPIC) study (Phase 2); in Phase 3 the validation of the candidate biomarkers by targeted proteomics on 48 colorectal cancer cases and 48 matched controls from the Florence-EPIC cohort, and the evaluation of the disease risk were performed.
  
  
  RESULTS
  Systems biology tools indicated that both in the Phase 1 and Phase 2 studies circulating protein levels differing in cases more than 1.5 times from controls, were involved in inflammation and/or immune response. Eight proteins including apolipoprotein C-II, complement C4-B, complement component C9, clusterin, alpha-2-HS-glycoprotein, mannan-binding lectin serine-protease, mannose-binding protein C, and N-acetylmuramoyl-L-alanine amidase were selected as promising candidate biomarkers. Targeted proteomics of the selected proteins in the EPIC samples showed significantly higher clusterin levels in cases than controls, but only in men (mean ± SD, 1.98 ± 0.46 and 1.61 ± 0.43 nmol/mL respectively, Mann-Whitney U, two-tailed P = 0.0173). The remaining proteins were unchanged. Using multivariate logistic models a significant positive association emerged for clusterin, with an 80% increase in the colorectal cancer risk with protein's unit increase, but only in men.
  
  
  CONCLUSIONS
  The results show that plasma proteins can be altered years before colorectal cancer detection. The high circulating clusterin in pre-diagnostic samples suggests this biomarker can improve the identification of people at risk of colorectal cancer and might help in designing preventive interventions.
authors.affiliation: Department of Environmental Health Sciences, IRCCS-Istituto di Ricerche Farmacologiche Mario Negri, Via Giuseppe La Masa 19, 20156, Milano, Italy. michela.bertuzzi@teletu.it.
authors.forename: Michela
authors.initials: M
authors.lastname: Bertuzzi
authors.affiliation: Department of Environmental Health Sciences, IRCCS-Istituto di Ricerche Farmacologiche Mario Negri, Via Giuseppe La Masa 19, 20156, Milano, Italy. cristina_marelli@yahoo.it.
authors.forename: Cristina
authors.initials: C
authors.lastname: Marelli
authors.affiliation: Department of Environmental Health Sciences, IRCCS-Istituto di Ricerche Farmacologiche Mario Negri, Via Giuseppe La Masa 19, 20156, Milano, Italy. renzo.bagnati@marionegri.it.
authors.forename: Renzo
authors.initials: R
authors.lastname: Bagnati
authors.affiliation: Department of Environmental Health Sciences, IRCCS-Istituto di Ricerche Farmacologiche Mario Negri, Via Giuseppe La Masa 19, 20156, Milano, Italy. co02ale@yahoo.it.
authors.forename: Alessandro
authors.initials: A
authors.lastname: Colombi
authors.affiliation: Department of Environmental Health Sciences, IRCCS-Istituto di Ricerche Farmacologiche Mario Negri, Via Giuseppe La Masa 19, 20156, Milano, Italy. 1roberto.fanelli@marionegri.it.
authors.forename: Roberto
authors.initials: R
authors.lastname: Fanelli
authors.affiliation: Molecular and Nutritional Epidemiology Unit, ISPO - Cancer Research and Prevention Institute, Florence, Italy. c.saieva@ispo.toscana.it.
authors.forename: Calogero
authors.initials: C
authors.lastname: Saieva
authors.affiliation: Molecular and Nutritional Epidemiology Unit, ISPO - Cancer Research and Prevention Institute, Florence, Italy. m.ceroti@ispo.toscana.it.
authors.forename: Marco
authors.initials: M
authors.lastname: Ceroti
authors.affiliation: Molecular and Nutritional Epidemiology Unit, ISPO - Cancer Research and Prevention Institute, Florence, Italy. b.bendinelli@ispo.toscana.it.
authors.forename: Benedetta
authors.initials: B
authors.lastname: Bendinelli
authors.affiliation: Molecular and Nutritional Epidemiology Unit, ISPO - Cancer Research and Prevention Institute, Florence, Italy. s.caini@ispo.toscana.it.
authors.forename: Saverio
authors.initials: S
authors.lastname: Caini
authors.affiliation: Department of Environmental Health Sciences, IRCCS-Istituto di Ricerche Farmacologiche Mario Negri, Via Giuseppe La Masa 19, 20156, Milano, Italy. luisa.airoldi@marionegri.it.
authors.forename: Luisa
authors.initials: L
authors.lastname: Airoldi
authors.affiliation: Molecular and Nutritional Epidemiology Unit, ISPO - Cancer Research and Prevention Institute, Florence, Italy. d.palli@ispo.toscana.it.
authors.forename: Domenico
authors.initials: D
authors.lastname: Palli
mesh_terms: D000368:Aged; D014408:Biomarkers, Tumor / Q000097:blood*; D016022:Case-Control Studies; D051152:Clusterin / Q000097:blood*; D015179:Colorectal Neoplasms / Q000097:blood / Q000175:diagnosis*; D005260:Female; D006801:Humans; D007558:Italy; D008297:Male; D013058:Mass Spectrometry / Q000379:methods*; D008875:Middle Aged; D010865:Pilot Projects; D011446:Prospective Studies; D020543:Proteome / Q000378:metabolism; D040901:Proteomics / Q000379:methods*; D012307:Risk Factors; D012737:Sex Factors
chemical_list: D014408:Biomarkers, Tumor; C495242:CLU protein, human; D051152:Clusterin; D020543:Proteome