title: Novel highly specific anti-periostin antibodies uncover the functional importance of the fascilin 1-1 domain and highlight preferential expression of periostin in aggressive breast cancer.
abstract: Periostin (POSTN), a secreted homodimeric protein that binds integrins αvβ3, αvβ5, and α6β4, was originally found to be expressed in fetal tissues and in the adult upon injury particularly bone fractures due to its role in remodelling and repair. Recently it was found to be over-expressed in human breast cancer and a variety of other tumour types including head and neck squamous cell carcinoma, where its overexpression correlates with increased tumour invasion. Progress in studying its functional role in tumour pathogenesis has been hampered by the paucity of antibodies for its specific and sensitive detection. It has proven very difficult to obtain monoclonal antibodies (mAbs) against this highly conserved protein but we report here that combining infection of mice with lactate dehydrogenase elevating virus (LDV), a B cell activating arterivirus, with conjugation of human POSTN to ovalbumin as an immunogenic carrier, enabled us to develop six mAbs recognizing both human and mouse POSTN and inhibiting its binding to αvβ3 integrin. Two of the mAbs, MPB4B1 and MPC5B4, were tested and found to inhibit POSTN-induced migration of human endothelial colony forming cells. All six mAbs recognized amino acids 136-51 (APSNEAWDNLDSDIRR) within the POSTN fascilin (FAS) 1-1 domain revealing the functional importance of this motif; this was further highlighted by the ability of aa 136-151 peptide to inhibit integrin-mediated cell migration. Immunohistochemistry using MPC5B4, indicated that breast tumour cell POSTN expression was a strong prognostic indicator, along with tumour size, lymph node, and human epidermal growth factor receptor 2 (HER2) status.
authors.affiliation: University of Oxford Branch, Ludwig Cancer Research, Oxford, United Kingdom.|Department of Neurodegenerative Disease, UCL Institute of Neurology, Queen Square, London, United Kingdom.
authors.forename: Sarah
authors.initials: S
authors.lastname: Field
authors.affiliation: Ludwig Cancer Research, Brussels Branch, Brussels, Belgium.|de Duve Institute, Université Catholique De Louvain, Brussels, Belgium.
authors.forename: Catherine
authors.initials: C
authors.lastname: Uyttenhove
authors.affiliation: Ludwig Cancer Research, Brussels Branch, Brussels, Belgium.
authors.forename: Vincent
authors.initials: V
authors.lastname: Stroobant
authors.affiliation: de Duve Institute, Université Catholique De Louvain, Brussels, Belgium.
authors.forename: Paméla
authors.initials: P
authors.lastname: Cheou
authors.affiliation: de Duve Institute, Université Catholique De Louvain, Brussels, Belgium.
authors.forename: Dominique
authors.initials: D
authors.lastname: Donckers
authors.affiliation: de Duve Institute, Université Catholique De Louvain, Brussels, Belgium.
authors.forename: Jean-Paul
authors.initials: JP
authors.lastname: Coutelier
authors.affiliation: The University of Queensland, UQ Centre for Clinical Research, Herston, Brisbane, Australia.|Cancer Genetics Laboratory, QIMR Berghofer Medical Research Institute, Queensland, Herston, Australia.|The University of Queensland, School of Medicine, Discipline of Molecular & Cellular Pathology, Herston, Brisbane, Australia.
authors.forename: Peter T
authors.initials: PT
authors.lastname: Simpson
authors.affiliation: The University of Queensland, UQ Centre for Clinical Research, Herston, Brisbane, Australia.|The University of Queensland, School of Medicine, Discipline of Molecular & Cellular Pathology, Herston, Brisbane, Australia.|Pathology Queensland, The Royal Brisbane & Women's Hospital, Brisbane, Australia.
authors.forename: Margaret C
authors.initials: MC
authors.lastname: Cummings
authors.affiliation: The University of Queensland, UQ Centre for Clinical Research, Herston, Brisbane, Australia.|Cancer Genetics Laboratory, QIMR Berghofer Medical Research Institute, Queensland, Herston, Australia.
authors.forename: Jodi M
authors.initials: JM
authors.lastname: Saunus
authors.affiliation: The University of Queensland, UQ Centre for Clinical Research, Herston, Brisbane, Australia.|Cancer Genetics Laboratory, QIMR Berghofer Medical Research Institute, Queensland, Herston, Australia.
authors.forename: Lynne E
authors.initials: LE
authors.lastname: Reid
authors.affiliation: The University of Queensland, UQ Centre for Clinical Research, Herston, Brisbane, Australia.|Cancer Genetics Laboratory, QIMR Berghofer Medical Research Institute, Queensland, Herston, Australia.
authors.forename: Jamie R
authors.initials: JR
authors.lastname: Kutasovic
authors.affiliation: The University of Queensland, UQ Centre for Clinical Research, Herston, Brisbane, Australia.|The University of Queensland, School of Medicine, Discipline of Molecular & Cellular Pathology, Herston, Brisbane, Australia.
authors.forename: Anne Marie
authors.initials: AM
authors.lastname: McNicol
authors.affiliation: Medical Research Centre for Ischemic Tissue Regeneration, School of Medicine, Pusan National University, Yangsan, Gyeongsangnam-do, Republic of Korea.|Department of Physiology, School of Medicine, Pusan National University, Yangsan, Gyeongsangnam-do, Republic of Korea.
authors.forename: Ba Reun
authors.initials: BR
authors.lastname: Kim
authors.affiliation: Medical Research Centre for Ischemic Tissue Regeneration, School of Medicine, Pusan National University, Yangsan, Gyeongsangnam-do, Republic of Korea.|Department of Physiology, School of Medicine, Pusan National University, Yangsan, Gyeongsangnam-do, Republic of Korea.|Research Institute of Convergence Biomedical Science and Technology, Pusan National University Yangsan Hospital, Yangsan, Gyeongsangnam-do, Republic of Korea.
authors.forename: Jae Ho
authors.initials: JH
authors.lastname: Kim
authors.affiliation: The University of Queensland, UQ Centre for Clinical Research, Herston, Brisbane, Australia.|The University of Queensland, School of Medicine, Discipline of Molecular & Cellular Pathology, Herston, Brisbane, Australia.|Pathology Queensland, The Royal Brisbane & Women's Hospital, Brisbane, Australia.
authors.forename: Sunil R
authors.initials: SR
authors.lastname: Lakhani
authors.affiliation: Ludwig Cancer Research, New York, NY.
authors.forename: A Munro
authors.initials: AM
authors.lastname: Neville
authors.affiliation: Ludwig Cancer Research, Brussels Branch, Brussels, Belgium.|de Duve Institute, Université Catholique De Louvain, Brussels, Belgium.
authors.forename: Jacques
authors.initials: J
authors.lastname: Van Snick
authors.affiliation: Department of Neurodegenerative Disease, UCL Institute of Neurology, Queen Square, London, United Kingdom.|MRC Prion Unit, UCL Institute of Neurology, Queen Square, London, United Kingdom.
authors.forename: Parmjit S
authors.identifier: http://orcid.org/0000-0002-3354-0594
authors.initials: PS
authors.lastname: Jat
mesh_terms: D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D020816:Amino Acid Motifs; D000818:Animals; D000911:Antibodies, Monoclonal; D000918:Antibody Specificity; D001666:Binding Sites, Antibody; D014408:Biomarkers, Tumor / Q000032:analysis*; D001943:Breast Neoplasms / Q000378:metabolism / Q000473:pathology*; D015815:Cell Adhesion Molecules / Q000378:metabolism*; D002465:Cell Movement / Q000502:physiology; D005260:Female; D006801:Humans; D007150:Immunohistochemistry; D051379:Mice; D008875:Middle Aged; D046888:Tissue Array Analysis
chemical_list: D000911:Antibodies, Monoclonal; D014408:Biomarkers, Tumor; D015815:Cell Adhesion Molecules; C512835:POSTN protein, human
keywords: FAS1-1 domain; breast cancer; diagnostic marker; extracellular matrix protein; monoclonal antibodies; periostin