title: Inhibition of TGFβ signaling decreases osteogenic differentiation of fibrodysplasia ossificans progressiva fibroblasts in a novel in vitro model of the disease.
abstract: Fibrodysplasia ossificans progressiva is a rare genetic disorder characterized by progressive heterotopic ossification. FOP patients develop soft tissue lumps as a result of inflammation-induced flare-ups which leads to the irreversible replacement of skeletal muscle tissue with bone tissue. Classical FOP patients possess a mutation (c.617G>A; R206H) in the ACVR1-encoding gene which leads to dysregulated BMP signaling. Nonetheless, not all FOP patients with this mutation exhibit equal severity in symptom presentation or disease progression which indicates a strong contribution by environmental factors. Given the pro-inflammatory role of TGFβ, we studied the role of TGFβ in the progression of osteogenic differentiation in primary dermal fibroblasts from five classical FOP patients based on a novel method of platelet lysate-based osteogenic transdifferentiation. During the course of transdifferentiation the osteogenic properties of the cells were evaluated by the mRNA expression of Sp7/Osterix, Runx2, Alp, OC and the presence of mineralization. During transdifferentiation the expression of osteoblast markers Runx2 (p<0.05) and Alp were higher in patient cells compared to healthy controls. All cell lines exhibited increase in mineralisation. FOP fibroblasts also expressed higher baseline Sp7/Osterix levels (p<0.05) confirming their higher osteogenic potential. The pharmacological inhibition of TGFβ signaling during osteogenic transdifferentiation resulted in the attenuation of osteogenic transdifferentiation in all cell lines as shown by the decrease in the expression of Runx2 (p<0.05), Alp and mineralization. We suggest that blocking of TGFβ signaling can decrease the osteogenic transdifferentiation of FOP fibroblasts.
authors.affiliation: Department of Clinical Genetics, VU University Medical Center, Amsterdam, The Netherlands. Electronic address: d.micha@vumc.nl.
authors.forename: Dimitra
authors.initials: D
authors.lastname: Micha
authors.affiliation: Department of Clinical Genetics, VU University Medical Center, Amsterdam, The Netherlands. Electronic address: elisevoermans@hotmail.com.
authors.forename: Elise
authors.initials: E
authors.lastname: Voermans
authors.affiliation: Internal Medicine, Endocrinology Section, VU University Medical Center, Amsterdam, The Netherlands. Electronic address: EMW.Eekhoff@vumc.nl.
authors.forename: Marelise E W
authors.initials: MEW
authors.lastname: Eekhoff
authors.affiliation: Department of Clinical Chemistry, VU University Medical Center, MOVE Research Institute, Amsterdam, The Netherlands. Electronic address: HW.vanEssen@vumc.nl.
authors.forename: Huib W
authors.initials: HW
authors.lastname: van Essen
authors.affiliation: Department of Oral Cell Biology, Academic Centre for Dentistry Amsterdam (ACTA), University of Amsterdam and VU University, MOVE Research Institute, Amsterdam, The Netherlands. Electronic address: bzandiehdoulabi@acta.nl.
authors.forename: Behrouz
authors.initials: B
authors.lastname: Zandieh-Doulabi
authors.affiliation: Internal Medicine, Endocrinology Section, VU University Medical Center, Amsterdam, The Netherlands. Electronic address: cnetelen@planet.nl.
authors.forename: Coen
authors.initials: C
authors.lastname: Netelenbos
authors.affiliation: Department of Dermatology, VU University Medical Centre, Amsterdam, The Netherlands. Electronic address: t.rustemeyer@vumc.nl.
authors.forename: Thomas
authors.initials: T
authors.lastname: Rustemeyer
authors.affiliation: Department of Clinical Genetics, VU University Medical Center, Amsterdam, The Netherlands. Electronic address: e.sistermans@vumc.nl.
authors.forename: E A
authors.initials: EA
authors.lastname: Sistermans
authors.affiliation: Department of Clinical Genetics, VU University Medical Center, Amsterdam, The Netherlands. Electronic address: g.pals@vumc.nl.
authors.forename: Gerard
authors.initials: G
authors.lastname: Pals
authors.affiliation: Department of Clinical Chemistry, VU University Medical Center, MOVE Research Institute, Amsterdam, The Netherlands. Electronic address: n.bravenboer@vumc.nl.
authors.forename: Nathalie
authors.initials: N
authors.lastname: Bravenboer
mesh_terms: D030201:Activin Receptors, Type I / Q000378:metabolism; D000293:Adolescent; D000328:Adult; D000469:Alkaline Phosphatase / Q000378:metabolism; D000818:Animals; D001549:Benzamides / Q000494:pharmacology; D001792:Blood Platelets / Q000378:metabolism; D002113:Calcification, Physiologic / Q000187:drug effects / Q000235:genetics; D054338:Cell Transdifferentiation / Q000187:drug effects / Q000235:genetics; D002478:Cells, Cultured; D005260:Female; D005347:Fibroblasts / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D005786:Gene Expression Regulation / Q000187:drug effects; D006801:Humans; D019394:Ki-67 Antigen / Q000378:metabolism; D008297:Male; D008875:Middle Aged; D008954:Models, Biological; D009221:Myositis Ossificans / Q000235:genetics / Q000378:metabolism* / Q000473:pathology*; D010012:Osteogenesis / Q000187:drug effects / Q000235:genetics; D011720:Pyrazoles / Q000494:pharmacology; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects / Q000235:genetics; D051785:Smad Proteins / Q000378:metabolism; D016212:Transforming Growth Factor beta / Q000378:metabolism*; D055815:Young Adult
chemical_list: C509927:4-(4-(3-(pyridin-2-yl)-1H-pyrazol-4-yl)pyridin-2-yl)-N-(tetrahydro-2H-pyran-4-yl)benzamide; D001549:Benzamides; D019394:Ki-67 Antigen; D011720:Pyrazoles; D012333:RNA, Messenger; D051785:Smad Proteins; D016212:Transforming Growth Factor beta; D030201:Activin Receptors, Type I; D000469:Alkaline Phosphatase
keywords: Fibroblast; Fibrodyplasia ossificans progressiva; Inflammation; Ossification; Osteogenic differentiation; Transforming growth factor β