title: High-throughput screening for modulators of ACVR1 transcription: discovery of potential therapeutics for fibrodysplasia ossificans progressiva.
abstract: The ACVR1 gene encodes a type I receptor of bone morphogenetic proteins (BMPs). Activating mutations in ACVR1 are responsible for fibrodysplasia ossificans progressiva (FOP), a rare disease characterized by congenital toe malformation and progressive heterotopic endochondral ossification leading to severe and cumulative disability. Until now, no therapy has been available to prevent soft-tissue swelling (flare-ups) that trigger the ossification process. With the aim of finding a new therapeutic strategy for FOP, we developed a high-throughput screening (HTS) assay to identify inhibitors of ACVR1 gene expression among drugs already approved for the therapy of other diseases. The screening, based on an ACVR1 promoter assay, was followed by an in vitro and in vivo test to validate and characterize candidate molecules. Among compounds that modulate the ACVR1 promoter activity, we selected the one showing the highest inhibitory effect, dipyridamole, a drug that is currently used as a platelet anti-aggregant. The inhibitory effect was detectable on ACVR1 gene expression, on the whole Smad-dependent BMP signaling pathway, and on chondrogenic and osteogenic differentiation processes by in vitro cellular assays. Moreover, dipyridamole reduced the process of heterotopic bone formation in vivo Our drug repositioning strategy has led to the identification of dipyridamole as a possible therapeutic tool for the treatment of FOP. Furthermore, our study has also defined a pipeline of assays that will be useful for the evaluation of other pharmacological inhibitors of heterotopic ossification.
authors.affiliation: Department of Neurosciences, Rehabilitation, Ophthalmology, Genetics, Maternal and Child Health and CEBR, Università degli Studi di Genova, Genova 16132, Italy.
authors.forename: Serena
authors.initials: S
authors.lastname: Cappato
authors.affiliation: Department of Neurosciences, Rehabilitation, Ophthalmology, Genetics, Maternal and Child Health and CEBR, Università degli Studi di Genova, Genova 16132, Italy.
authors.forename: Laura
authors.initials: L
authors.lastname: Tonachini
authors.affiliation: Department of Neurosciences, Rehabilitation, Ophthalmology, Genetics, Maternal and Child Health and CEBR, Università degli Studi di Genova, Genova 16132, Italy.
authors.forename: Francesca
authors.initials: F
authors.lastname: Giacopelli
authors.affiliation: Division of Immunology, Transplantation and Infectious Diseases, San Raffaele Scientific Institute, Milano 20132, Italy School of Medicine and Surgery, University of Milano-Bicocca, Monza 20900, Italy.
authors.forename: Mario
authors.initials: M
authors.lastname: Tirone
authors.affiliation: Medical Genetics Unit, IRCCS Istituto Giannina Gaslini, Genova 16147, Italy.
authors.forename: Luis J V
authors.initials: LJ
authors.lastname: Galietta
authors.affiliation: School of Medicine and Surgery, University of Milano-Bicocca, Monza 20900, Italy.
authors.forename: Martina
authors.initials: M
authors.lastname: Sormani
authors.affiliation: School of Medicine and Surgery, University of Milano-Bicocca, Monza 20900, Italy.
authors.forename: Anna
authors.initials: A
authors.lastname: Giovenzana
authors.affiliation: Medical Physics Department and Centre for Experimental Imaging, San Raffaele Scientific Institute, Milano 20132, Italy.
authors.forename: Antonello E
authors.initials: AE
authors.lastname: Spinelli
authors.affiliation: Dipartimento di Medicina Sperimentale, Università di Genova & IRCCS AOU San Martino-IST, Istituto Nazionale per la Ricerca sul Cancro, 16132 Genova, Italy.
authors.forename: Barbara
authors.initials: B
authors.lastname: Canciani
authors.affiliation: School of Medicine and Surgery, University of Milano-Bicocca, Monza 20900, Italy.
authors.forename: Silvia
authors.initials: S
authors.lastname: Brunelli
authors.affiliation: Department of Neurosciences, Rehabilitation, Ophthalmology, Genetics, Maternal and Child Health and CEBR, Università degli Studi di Genova, Genova 16132, Italy Medical Genetics Unit, IRCCS Istituto Giannina Gaslini, Genova 16147, Italy.
authors.forename: Roberto
authors.initials: R
authors.lastname: Ravazzolo
authors.affiliation: Department of Neurosciences, Rehabilitation, Ophthalmology, Genetics, Maternal and Child Health and CEBR, Università degli Studi di Genova, Genova 16132, Italy Medical Genetics Unit, IRCCS Istituto Giannina Gaslini, Genova 16147, Italy bocciardi@unige.it.
authors.forename: Renata
authors.identifier: 0000-0001-8415-3802
authors.initials: R
authors.lastname: Bocciardi
mesh_terms: D030201:Activin Receptors, Type I / Q000235:genetics*; D000818:Animals; D015415:Biomarkers / Q000378:metabolism; D019485:Bone Morphogenetic Proteins / Q000378:metabolism; D002118:Calcium / Q000378:metabolism; D002454:Cell Differentiation / Q000187:drug effects; D002460:Cell Line; D020219:Chondrogenesis / Q000187:drug effects; D004176:Dipyridamole / Q000494:pharmacology / Q000627:therapeutic use; D004195:Disease Models, Animal; D057166:High-Throughput Screening Assays / Q000379:methods*; D051379:Mice; D009221:Myositis Ossificans / Q000188:drug therapy* / Q000378:metabolism / Q000473:pathology; D009999:Ossification, Heterotopic / Q000000981:diagnostic imaging / Q000473:pathology; D010006:Osteoblasts / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D010012:Osteogenesis / Q000187:drug effects; D015203:Reproducibility of Results; D015398:Signal Transduction / Q000187:drug effects; D051785:Smad Proteins / Q000378:metabolism; D014158:Transcription, Genetic / Q000187:drug effects
chemical_list: D015415:Biomarkers; D019485:Bone Morphogenetic Proteins; D051785:Smad Proteins; D004176:Dipyridamole; D030201:Activin Receptors, Type I; D002118:Calcium
keywords: ACVR1; BMP signaling pathway; Dipyridamole; Drug repositioning; FOP; High-throughput screening; Transcriptional regulation