title: Tumour-associated fibroblasts contribute to neoangiogenesis in human parathyroid neoplasia.
abstract: Components of the tumour microenvironment initiate and promote cancer development. In this study, we investigated the stromal component of parathyroid neoplasia. Immunohistochemistry for alpha-smooth muscle actin (α-SMA) showed an abundant periacinar distribution of α-SMA(+) cells in normal parathyroid glands (n=3). This pattern was progressively lost in parathyroid adenomas (PAds; n=6) where α-SMA(+)cells were found to surround new microvessels, as observed in foetal parathyroid glands (n=2). Moreover, in atypical adenomas (n=5) and carcinomas (n=4), α-SMA(+) cells disappeared from the parenchyma and accumulated in the capsula and fibrous bands. At variance with normal glands, parathyroid tumours (n=37) expressed high levels of fibroblast-activation protein (FAP) transcripts, a marker of tumour-associated fibroblasts. We analysed the ability of PAd-derived cells to activate fibroblasts using human bone-marrow mesenchymal stem cells (hBM-MSCs). PAd-derived cells induced a significant increase in FAP and vascular endothelial growth factor A (VEGFA) mRNA levels in co-cultured hBM-MSCs. Furthermore, the role of the calcium-sensing receptor (CASR) and of the CXCL12/CXCR4 pathway in the PAd-induced activation of hBM-MSCs was investigated. Treatment of co-cultures of hBM-MSCs and PAd-derived cells with the CXCR4 inhibitor AMD3100 reduced the stimulated VEGFA levels, while CASR activation by the R568 agonist was ineffective. PAd-derived cells co-expressing parathyroid hormone (PTH)/CXCR4 and PTH/CXCL12 were identified by FACS, suggesting a paracrine/autocrine signalling. Finally, CXCR4 blockade by AMD3100 reduced PTH gene expression levels in PAd-derived cells. In conclusion, i) PAd-derived cells activated cells of mesenchymal origin; ii) PAd-associated fibroblasts were involved in tumuor neoangiogenesis and iii) CXCL12/CXCR4 pathway was expressed and active in PAd cells, likely contributing to parathyroid tumour neoangiogenesis and PTH synthesis modulation.
authors.affiliation: Laboratory of Molecular BiologyIRCCS Policlinico San Donato, San Donato Milanese, Milan, ItalyDepartment of Human PathologySan Paolo Hospital, University of Milan, Milan, ItalyLaboratory of Stem Cells for Tissue EngineeringIRCCS Policlinico San Donato, San Donato Milanese, Milan, ItalyMedical GeneticsEndocrinology UnitIRCCS Hospital Casa Sollievo Sofferenza, San Giovanni Rotondo, Foggia, ItalyEndocrine SurgeryIRCCS Fondazione Cà Granda Ospedale Maggiore Policlinico, Milan, ItalySurgery UnitIRCCS Policlinico San Donato, San Donato Milanese, Milan, ItalySurgery UnitIRCCS Ospedale San Raffaele, Milan, ItalyDepartment of Health SciencesEndocrinology and Diabetology UnitDepartment of Clinical and Community Sciences, IRCCS Fondazione Cà Granda Ospedale Maggiore Policlinico, University of Milan, Milan, ItalyEndocrinology and Diabetology UnitDepartment of Biomedical Sciences for Health, IRCCS Policlinico San Donato, University of Milan, Via Morandi 30, 20097 San Donato Milanese, Milan, Italy.
authors.forename: C
authors.initials: C
authors.lastname: Verdelli
authors.affiliation: Laboratory of Molecular BiologyIRCCS Policlinico San Donato, San Donato Milanese, Milan, ItalyDepartment of Human PathologySan Paolo Hospital, University of Milan, Milan, ItalyLaboratory of Stem Cells for Tissue EngineeringIRCCS Policlinico San Donato, San Donato Milanese, Milan, ItalyMedical GeneticsEndocrinology UnitIRCCS Hospital Casa Sollievo Sofferenza, San Giovanni Rotondo, Foggia, ItalyEndocrine SurgeryIRCCS Fondazione Cà Granda Ospedale Maggiore Policlinico, Milan, ItalySurgery UnitIRCCS Policlinico San Donato, San Donato Milanese, Milan, ItalySurgery UnitIRCCS Ospedale San Raffaele, Milan, ItalyDepartment of Health SciencesEndocrinology and Diabetology UnitDepartment of Clinical and Community Sciences, IRCCS Fondazione Cà Granda Ospedale Maggiore Policlinico, University of Milan, Milan, ItalyEndocrinology and Diabetology UnitDepartment of Biomedical Sciences for Health, IRCCS Policlinico San Donato, University of Milan, Via Morandi 30, 20097 San Donato Milanese, Milan, Italy.
authors.forename: L
authors.initials: L
authors.lastname: Avagliano
authors.affiliation: Laboratory of Molecular BiologyIRCCS Policlinico San Donato, San Donato Milanese, Milan, ItalyDepartment of Human PathologySan Paolo Hospital, University of Milan, Milan, ItalyLaboratory of Stem Cells for Tissue EngineeringIRCCS Policlinico San Donato, San Donato Milanese, Milan, ItalyMedical GeneticsEndocrinology UnitIRCCS Hospital Casa Sollievo Sofferenza, San Giovanni Rotondo, Foggia, ItalyEndocrine SurgeryIRCCS Fondazione Cà Granda Ospedale Maggiore Policlinico, Milan, ItalySurgery UnitIRCCS Policlinico San Donato, San Donato Milanese, Milan, ItalySurgery UnitIRCCS Ospedale San Raffaele, Milan, ItalyDepartment of Health SciencesEndocrinology and Diabetology UnitDepartment of Clinical and Community Sciences, IRCCS Fondazione Cà Granda Ospedale Maggiore Policlinico, University of Milan, Milan, ItalyEndocrinology and Diabetology UnitDepartment of Biomedical Sciences for Health, IRCCS Policlinico San Donato, University of Milan, Via Morandi 30, 20097 San Donato Milanese, Milan, Italy.
authors.forename: P
authors.initials: P
authors.lastname: Creo
authors.affiliation: Laboratory of Molecular BiologyIRCCS Policlinico San Donato, San Donato Milanese, Milan, ItalyDepartment of Human PathologySan Paolo Hospital, University of Milan, Milan, ItalyLaboratory of Stem Cells for Tissue EngineeringIRCCS Policlinico San Donato, San Donato Milanese, Milan, ItalyMedical GeneticsEndocrinology UnitIRCCS Hospital Casa Sollievo Sofferenza, San Giovanni Rotondo, Foggia, ItalyEndocrine SurgeryIRCCS Fondazione Cà Granda Ospedale Maggiore Policlinico, Milan, ItalySurgery UnitIRCCS Policlinico San Donato, San Donato Milanese, Milan, ItalySurgery UnitIRCCS Ospedale San Raffaele, Milan, ItalyDepartment of Health SciencesEndocrinology and Diabetology UnitDepartment of Clinical and Community Sciences, IRCCS Fondazione Cà Granda Ospedale Maggiore Policlinico, University of Milan, Milan, ItalyEndocrinology and Diabetology UnitDepartment of Biomedical Sciences for Health, IRCCS Policlinico San Donato, University of Milan, Via Morandi 30, 20097 San Donato Milanese, Milan, Italy.
authors.forename: V
authors.initials: V
authors.lastname: Guarnieri
authors.affiliation: Laboratory of Molecular BiologyIRCCS Policlinico San Donato, San Donato Milanese, Milan, ItalyDepartment of Human PathologySan Paolo Hospital, University of Milan, Milan, ItalyLaboratory of Stem Cells for Tissue EngineeringIRCCS Policlinico San Donato, San Donato Milanese, Milan, ItalyMedical GeneticsEndocrinology UnitIRCCS Hospital Casa Sollievo Sofferenza, San Giovanni Rotondo, Foggia, ItalyEndocrine SurgeryIRCCS Fondazione Cà Granda Ospedale Maggiore Policlinico, Milan, ItalySurgery UnitIRCCS Policlinico San Donato, San Donato Milanese, Milan, ItalySurgery UnitIRCCS Ospedale San Raffaele, Milan, ItalyDepartment of Health SciencesEndocrinology and Diabetology UnitDepartment of Clinical and Community Sciences, IRCCS Fondazione Cà Granda Ospedale Maggiore Policlinico, University of Milan, Milan, ItalyEndocrinology and Diabetology UnitDepartment of Biomedical Sciences for Health, IRCCS Policlinico San Donato, University of Milan, Via Morandi 30, 20097 San Donato Milanese, Milan, Italy.
authors.forename: A
authors.initials: A
authors.lastname: Scillitani
authors.affiliation: Laboratory of Molecular BiologyIRCCS Policlinico San Donato, San Donato Milanese, Milan, ItalyDepartment of Human PathologySan Paolo Hospital, University of Milan, Milan, ItalyLaboratory of Stem Cells for Tissue EngineeringIRCCS Policlinico San Donato, San Donato Milanese, Milan, ItalyMedical GeneticsEndocrinology UnitIRCCS Hospital Casa Sollievo Sofferenza, San Giovanni Rotondo, Foggia, ItalyEndocrine SurgeryIRCCS Fondazione Cà Granda Ospedale Maggiore Policlinico, Milan, ItalySurgery UnitIRCCS Policlinico San Donato, San Donato Milanese, Milan, ItalySurgery UnitIRCCS Ospedale San Raffaele, Milan, ItalyDepartment of Health SciencesEndocrinology and Diabetology UnitDepartment of Clinical and Community Sciences, IRCCS Fondazione Cà Granda Ospedale Maggiore Policlinico, University of Milan, Milan, ItalyEndocrinology and Diabetology UnitDepartment of Biomedical Sciences for Health, IRCCS Policlinico San Donato, University of Milan, Via Morandi 30, 20097 San Donato Milanese, Milan, Italy.
authors.forename: L
authors.initials: L
authors.lastname: Vicentini
authors.affiliation: Laboratory of Molecular BiologyIRCCS Policlinico San Donato, San Donato Milanese, Milan, ItalyDepartment of Human PathologySan Paolo Hospital, University of Milan, Milan, ItalyLaboratory of Stem Cells for Tissue EngineeringIRCCS Policlinico San Donato, San Donato Milanese, Milan, ItalyMedical GeneticsEndocrinology UnitIRCCS Hospital Casa Sollievo Sofferenza, San Giovanni Rotondo, Foggia, ItalyEndocrine SurgeryIRCCS Fondazione Cà Granda Ospedale Maggiore Policlinico, Milan, ItalySurgery UnitIRCCS Policlinico San Donato, San Donato Milanese, Milan, ItalySurgery UnitIRCCS Ospedale San Raffaele, Milan, ItalyDepartment of Health SciencesEndocrinology and Diabetology UnitDepartment of Clinical and Community Sciences, IRCCS Fondazione Cà Granda Ospedale Maggiore Policlinico, University of Milan, Milan, ItalyEndocrinology and Diabetology UnitDepartment of Biomedical Sciences for Health, IRCCS Policlinico San Donato, University of Milan, Via Morandi 30, 20097 San Donato Milanese, Milan, Italy.
authors.forename: G B
authors.initials: GB
authors.lastname: Steffano
authors.affiliation: Laboratory of Molecular BiologyIRCCS Policlinico San Donato, San Donato Milanese, Milan, ItalyDepartment of Human PathologySan Paolo Hospital, University of Milan, Milan, ItalyLaboratory of Stem Cells for Tissue EngineeringIRCCS Policlinico San Donato, San Donato Milanese, Milan, ItalyMedical GeneticsEndocrinology UnitIRCCS Hospital Casa Sollievo Sofferenza, San Giovanni Rotondo, Foggia, ItalyEndocrine SurgeryIRCCS Fondazione Cà Granda Ospedale Maggiore Policlinico, Milan, ItalySurgery UnitIRCCS Policlinico San Donato, San Donato Milanese, Milan, ItalySurgery UnitIRCCS Ospedale San Raffaele, Milan, ItalyDepartment of Health SciencesEndocrinology and Diabetology UnitDepartment of Clinical and Community Sciences, IRCCS Fondazione Cà Granda Ospedale Maggiore Policlinico, University of Milan, Milan, ItalyEndocrinology and Diabetology UnitDepartment of Biomedical Sciences for Health, IRCCS Policlinico San Donato, University of Milan, Via Morandi 30, 20097 San Donato Milanese, Milan, Italy.
authors.forename: E
authors.initials: E
authors.lastname: Beretta
authors.affiliation: Laboratory of Molecular BiologyIRCCS Policlinico San Donato, San Donato Milanese, Milan, ItalyDepartment of Human PathologySan Paolo Hospital, University of Milan, Milan, ItalyLaboratory of Stem Cells for Tissue EngineeringIRCCS Policlinico San Donato, San Donato Milanese, Milan, ItalyMedical GeneticsEndocrinology UnitIRCCS Hospital Casa Sollievo Sofferenza, San Giovanni Rotondo, Foggia, ItalyEndocrine SurgeryIRCCS Fondazione Cà Granda Ospedale Maggiore Policlinico, Milan, ItalySurgery UnitIRCCS Policlinico San Donato, San Donato Milanese, Milan, ItalySurgery UnitIRCCS Ospedale San Raffaele, Milan, ItalyDepartment of Health SciencesEndocrinology and Diabetology UnitDepartment of Clinical and Community Sciences, IRCCS Fondazione Cà Granda Ospedale Maggiore Policlinico, University of Milan, Milan, ItalyEndocrinology and Diabetology UnitDepartment of Biomedical Sciences for Health, IRCCS Policlinico San Donato, University of Milan, Via Morandi 30, 20097 San Donato Milanese, Milan, Italy.
authors.forename: L
authors.initials: L
authors.lastname: Soldati
authors.affiliation: Laboratory of Molecular BiologyIRCCS Policlinico San Donato, San Donato Milanese, Milan, ItalyDepartment of Human PathologySan Paolo Hospital, University of Milan, Milan, ItalyLaboratory of Stem Cells for Tissue EngineeringIRCCS Policlinico San Donato, San Donato Milanese, Milan, ItalyMedical GeneticsEndocrinology UnitIRCCS Hospital Casa Sollievo Sofferenza, San Giovanni Rotondo, Foggia, ItalyEndocrine SurgeryIRCCS Fondazione Cà Granda Ospedale Maggiore Policlinico, Milan, ItalySurgery UnitIRCCS Policlinico San Donato, San Donato Milanese, Milan, ItalySurgery UnitIRCCS Ospedale San Raffaele, Milan, ItalyDepartment of Health SciencesEndocrinology and Diabetology UnitDepartment of Clinical and Community Sciences, IRCCS Fondazione Cà Granda Ospedale Maggiore Policlinico, University of Milan, Milan, ItalyEndocrinology and Diabetology UnitDepartment of Biomedical Sciences for Health, IRCCS Policlinico San Donato, University of Milan, Via Morandi 30, 20097 San Donato Milanese, Milan, Italy.
authors.forename: E
authors.initials: E
authors.lastname: Costa
authors.affiliation: Laboratory of Molecular BiologyIRCCS Policlinico San Donato, San Donato Milanese, Milan, ItalyDepartment of Human PathologySan Paolo Hospital, University of Milan, Milan, ItalyLaboratory of Stem Cells for Tissue EngineeringIRCCS Policlinico San Donato, San Donato Milanese, Milan, ItalyMedical GeneticsEndocrinology UnitIRCCS Hospital Casa Sollievo Sofferenza, San Giovanni Rotondo, Foggia, ItalyEndocrine SurgeryIRCCS Fondazione Cà Granda Ospedale Maggiore Policlinico, Milan, ItalySurgery UnitIRCCS Policlinico San Donato, San Donato Milanese, Milan, ItalySurgery UnitIRCCS Ospedale San Raffaele, Milan, ItalyDepartment of Health SciencesEndocrinology and Diabetology UnitDepartment of Clinical and Community Sciences, IRCCS Fondazione Cà Granda Ospedale Maggiore Policlinico, University of Milan, Milan, ItalyEndocrinology and Diabetology UnitDepartment of Biomedical Sciences for Health, IRCCS Policlinico San Donato, University of Milan, Via Morandi 30, 20097 San Donato Milanese, Milan, Italy.
authors.forename: A
authors.initials: A
authors.lastname: Spada
authors.affiliation: Laboratory of Molecular BiologyIRCCS Policlinico San Donato, San Donato Milanese, Milan, ItalyDepartment of Human PathologySan Paolo Hospital, University of Milan, Milan, ItalyLaboratory of Stem Cells for Tissue EngineeringIRCCS Policlinico San Donato, San Donato Milanese, Milan, ItalyMedical GeneticsEndocrinology UnitIRCCS Hospital Casa Sollievo Sofferenza, San Giovanni Rotondo, Foggia, ItalyEndocrine SurgeryIRCCS Fondazione Cà Granda Ospedale Maggiore Policlinico, Milan, ItalySurgery UnitIRCCS Policlinico San Donato, San Donato Milanese, Milan, ItalySurgery UnitIRCCS Ospedale San Raffaele, Milan, ItalyDepartment of Health SciencesEndocrinology and Diabetology UnitDepartment of Clinical and Community Sciences, IRCCS Fondazione Cà Granda Ospedale Maggiore Policlinico, University of Milan, Milan, ItalyEndocrinology and Diabetology UnitDepartment of Biomedical Sciences for Health, IRCCS Policlinico San Donato, University of Milan, Via Morandi 30, 20097 San Donato Milanese, Milan, Italy.
authors.forename: G P
authors.initials: GP
authors.lastname: Bulfamante
authors.affiliation: Laboratory of Molecular BiologyIRCCS Policlinico San Donato, San Donato Milanese, Milan, ItalyDepartment of Human PathologySan Paolo Hospital, University of Milan, Milan, ItalyLaboratory of Stem Cells for Tissue EngineeringIRCCS Policlinico San Donato, San Donato Milanese, Milan, ItalyMedical GeneticsEndocrinology UnitIRCCS Hospital Casa Sollievo Sofferenza, San Giovanni Rotondo, Foggia, ItalyEndocrine SurgeryIRCCS Fondazione Cà Granda Ospedale Maggiore Policlinico, Milan, ItalySurgery UnitIRCCS Policlinico San Donato, San Donato Milanese, Milan, ItalySurgery UnitIRCCS Ospedale San Raffaele, Milan, ItalyDepartment of Health SciencesEndocrinology and Diabetology UnitDepartment of Clinical and Community Sciences, IRCCS Fondazione Cà Granda Ospedale Maggiore Policlinico, University of Milan, Milan, ItalyEndocrinology and Diabetology UnitDepartment of Biomedical Sciences for Health, IRCCS Policlinico San Donato, University of Milan, Via Morandi 30, 20097 San Donato Milanese, Milan, Italy sabrina.corbetta@unimi.it.
authors.forename: S
authors.initials: S
authors.lastname: Corbetta
mesh_terms: D000236:Adenoma / Q000098:blood supply* / Q000378:metabolism / Q000473:pathology*; D001596:Benzylamines; D018920:Coculture Techniques; D000080027:Cyclams; D005347:Fibroblasts / Q000187:drug effects / Q000378:metabolism / Q000473:pathology*; D015870:Gene Expression; D015972:Gene Expression Regulation, Neoplastic; D006571:Heterocyclic Compounds / Q000494:pharmacology; D006801:Humans; D007150:Immunohistochemistry; D059630:Mesenchymal Stem Cells / Q000473:pathology; D009389:Neovascularization, Pathologic / Q000188:drug therapy / Q000378:metabolism / Q000473:pathology; D010282:Parathyroid Neoplasms / Q000098:blood supply* / Q000378:metabolism / Q000473:pathology*; D015398:Signal Transduction; D017154:Stromal Cells / Q000473:pathology; D014407:Tumor Cells, Cultured; D059016:Tumor Microenvironment
chemical_list: D001596:Benzylamines; D000080027:Cyclams; D006571:Heterocyclic Compounds; C088327:plerixafor
keywords: CXCR4; PTH; SDF1/CXCL12; VEGFA; parathyroid; tumour-associated fibroblasts