title: Secretome profiling of oral squamous cell carcinoma-associated fibroblasts reveals organization and disassembly of extracellular matrix and collagen metabolic process signatures.
abstract: An important role has been attributed to cancer-associated fibroblasts (CAFs) in the tumorigenesis of oral squamous cell carcinoma (OSCC), the most common tumor of the oral cavity. Previous studies demonstrated that CAF-secreted molecules promote the proliferation and invasion of OSCC cells, inducing a more aggressive phenotype. In this study, we searched for differences in the secretome of CAFs and normal oral fibroblasts (NOF) using mass spectrometry-based proteomics and biological network analysis. Comparison of the secretome profiles revealed that upregulated proteins involved mainly in extracellular matrix organization and disassembly and collagen metabolism. Among the upregulated proteins were fibronectin type III domain-containing 1 (FNDC1), serpin peptidase inhibitor type 1 (SERPINE1), and stanniocalcin 2 (STC2), the upregulation of which was validated by quantitative PCR and ELISA in an independent set of CAF cell lines. The transition of transforming growth factor beta 1 (TGF-β1)-mediating NOFs into CAFs was accompanied by significant upregulation of FNDC1, SERPINE1, and STC2, confirming the participation of these proteins in the CAF-derived secretome. Type I collagen, the main constituent of the connective tissue, was also associated with several upregulated biological processes. The immunoexpression of type I collagen N-terminal propeptide (PINP) was significantly correlated in vivo with CAFs in the tumor front and was associated with significantly shortened survival of OSCC patients. Presence of CAFs in the tumor stroma was also an independent prognostic factor for OSCC disease-free survival. These results demonstrate the value of secretome profiling for evaluating the role of CAFs in the tumor microenvironment and identify potential novel therapeutic targets such as FNDC1, SERPINE1, and STC2. Furthermore, type I collagen expression by CAFs, represented by PINP levels, may be a prognostic marker of OSCC outcome.
authors.affiliation: Department of Oral Diagnosis, School of Dentistry, State University of Campinas, Av. Limeira 901, CEP 13414-018, Piracicaba, SP, Brazil.
authors.forename: Elizabete
authors.initials: E
authors.lastname: Bagordakis
authors.affiliation: Oral Pathology and Oral Medicine, Dentistry School, Western Paraná State University, Cascavel, PR, Brazil.
authors.forename: Iris
authors.initials: I
authors.lastname: Sawazaki-Calone
authors.affiliation: Department of Oral Diagnosis, School of Dentistry, State University of Campinas, Av. Limeira 901, CEP 13414-018, Piracicaba, SP, Brazil.
authors.forename: Carolina Carneiro Soares
authors.initials: CC
authors.lastname: Macedo
authors.affiliation: Brazilian Biociences National Laboratory-CNPEM, CEP 13083-970, Campinas, SP, Brazil.
authors.forename: Carolina M
authors.initials: CM
authors.lastname: Carnielli
authors.affiliation: Department of Oral Diagnosis, School of Dentistry, State University of Campinas, Av. Limeira 901, CEP 13414-018, Piracicaba, SP, Brazil.
authors.forename: Carine Ervolino
authors.initials: CE
authors.lastname: de Oliveira
authors.affiliation: Department of Oral Diagnosis, School of Dentistry, State University of Campinas, Av. Limeira 901, CEP 13414-018, Piracicaba, SP, Brazil.
authors.forename: Priscila Campioni
authors.initials: PC
authors.lastname: Rodrigues
authors.affiliation: Oral Pathology and Oral Medicine, Dentistry School, Western Paraná State University, Cascavel, PR, Brazil.
authors.forename: Ana Lucia C A
authors.initials: AL
authors.lastname: Rangel
authors.affiliation: Laboratory of Surgical Pathology, Dental School, Federal University of Bahia-UFBA, Salvador, BA, Brazil.
authors.forename: Jean Nunes
authors.initials: JN
authors.lastname: Dos Santos
authors.affiliation: Cancer and Translational Medicine Research Unit, University of Oulu, Oulu, Finland.|Medical Research Center, Oulu University Hospital, Oulu, Finland.
authors.forename: Juha
authors.initials: J
authors.lastname: Risteli
authors.affiliation: Department of Oral Diagnosis, School of Dentistry, State University of Campinas, Av. Limeira 901, CEP 13414-018, Piracicaba, SP, Brazil.
authors.forename: Edgard
authors.initials: E
authors.lastname: Graner
authors.affiliation: Cancer and Translational Medicine Research Unit, University of Oulu, Oulu, Finland.|Medical Research Center, Oulu University Hospital, Oulu, Finland.|Oral and Maxillofacial Diseases Unit, University of Helsinki, Helsinki, Finland.|Helsinki University Hospital, Helsinki, Finland.
authors.forename: Tuula
authors.initials: T
authors.lastname: Salo
authors.affiliation: Brazilian Biociences National Laboratory-CNPEM, CEP 13083-970, Campinas, SP, Brazil. adriana.paesleme@lnbio.cnpem.br.
authors.forename: Adriana Franco
authors.initials: AF
authors.lastname: Paes Leme
authors.affiliation: Department of Oral Diagnosis, School of Dentistry, State University of Campinas, Av. Limeira 901, CEP 13414-018, Piracicaba, SP, Brazil. coletta@fop.unicamp.br.
authors.forename: Ricardo D
authors.initials: RD
authors.lastname: Coletta
mesh_terms: D002294:Carcinoma, Squamous Cell / Q000378:metabolism* / Q000473:pathology; D002460:Cell Line; D045744:Cell Line, Tumor; D024042:Collagen Type I / Q000378:metabolism*; D018572:Disease-Free Survival; D005109:Extracellular Matrix / Q000378:metabolism* / Q000473:pathology; D005260:Female; D005347:Fibroblasts / Q000378:metabolism* / Q000473:pathology; D006023:Glycoproteins / Q000378:metabolism; D006801:Humans; D036341:Intercellular Signaling Peptides and Proteins / Q000378:metabolism; D008297:Male; D009062:Mouth Neoplasms / Q000378:metabolism* / Q000473:pathology; D009363:Neoplasm Proteins / Q000378:metabolism; D010446:Peptide Fragments / Q000378:metabolism; D017395:Plasminogen Activator Inhibitor 1 / Q000378:metabolism; D011347:Procollagen / Q000378:metabolism; D053773:Transforming Growth Factor beta1 / Q000378:metabolism; D059016:Tumor Microenvironment / Q000502:physiology; D015854:Up-Regulation / Q000502:physiology
chemical_list: D024042:Collagen Type I; C505811:FNDC1 protein, human; D006023:Glycoproteins; D036341:Intercellular Signaling Peptides and Proteins; D009363:Neoplasm Proteins; D010446:Peptide Fragments; D017395:Plasminogen Activator Inhibitor 1; D011347:Procollagen; C051321:SERPINE1 protein, human; C114997:STC2 protein, human; D053773:Transforming Growth Factor beta1; C047202:procollagen Type I N-terminal peptide
keywords: Cancer-associated fibroblasts; Extracellular matrix; FNDC1; SERPINE1; STC2; Secretome; Type I collagen