{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,9]],"date-time":"2025-10-09T00:43:44Z","timestamp":1759970624789,"version":"build-2065373602"},"reference-count":27,"publisher":"MDPI AG","issue":"3","license":[{"start":{"date-parts":[[2025,1,28]],"date-time":"2025-01-28T00:00:00Z","timestamp":1738022400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100005378","name":"Hacettepe University","doi-asserted-by":"publisher","award":["TSA-2021-19468"],"award-info":[{"award-number":["TSA-2021-19468"]}],"id":[{"id":"10.13039\/501100005378","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Skeletal dysplasias, characterized by bone, cartilage, and connective tissue abnormalities, often arise due to disruptions in extracellular matrix (ECM) dynamics and growth factor-dependent signaling pathways. RSPRY1, a secreted protein with RING and SPRY domains, has been implicated in bone development, yet its exact role remains to be determined. RSPRY1 gene mutations are associated with spondyloepimetaphyseal dysplasia (SEMD), a rare skeletal disorder characterized by severe epiphyseal and metaphyseal deformities. This study aimed to determine the molecular and cellular mechanisms by which RSPRY1 deficiency affects skeletal homeostasis. Transcriptome analysis of fibroblasts from patients with homozygous RSPRY1 mutations showed there was significant enrichment of transforming growth factor beta (TGF-\u03b2) signaling and ECM-related pathways. Functional wound healing assays showed that RSPRY1 knockout fibroblasts exhibited enhanced motility, a phenotype that was abrogated in RSPRY1 + SMAD3 double knockout fibroblasts, highlighting the SMAD3-dependence of RSPRY1\u2032s effects. The observed limited response to exogenous TGF-\u03b2 in RSPRY1-deficient cells indicated that there was constitutive pathway activation. These findings show that RSPRY1 is a critical regulator of TGF-\u03b2 signaling in ECM dynamics and cell motility, contributing to the pathophysiology of SEMD. An improvement in our understanding of the molecular roles of RSPRY1 might yield novel therapeutic strategies that target TGF-\u03b2 signaling in patients with SEMD and other skeletal dysplasias.<\/jats:p>","DOI":"10.3390\/ijms26031134","type":"journal-article","created":{"date-parts":[[2025,1,28]],"date-time":"2025-01-28T06:47:29Z","timestamp":1738046849000},"page":"1134","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Unraveling the Role of RSPRY1 in TGF-\u03b2 Pathway Dysregulation: Insights into the Pathogenesis of Spondyloepimetaphyseal Dysplasia"],"prefix":"10.3390","volume":"26","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-2556-0421","authenticated-orcid":false,"given":"Gozde","family":"Imren","sequence":"first","affiliation":[{"name":"Department of Medical Genetics, Faculty of Medicine, Hacettepe University, 06100 Ankara, T\u00fcrkiye"},{"name":"Department of Medical and Surgical Research, Institute of Health Sciences, Hacettepe University, 06100 Ankara, T\u00fcrkiye"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5564-4813","authenticated-orcid":false,"given":"Beren","family":"Karaosmanoglu","sequence":"additional","affiliation":[{"name":"Department of Medical Genetics, Faculty of Medicine, Hacettepe University, 06100 Ankara, T\u00fcrkiye"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0009-0001-2501-1668","authenticated-orcid":false,"given":"Bihter","family":"Muratoglu","sequence":"additional","affiliation":[{"name":"Department of Stem Cell Sciences, Institute of Health Sciences, Hacettepe University, 06100 Ankara, T\u00fcrkiye"},{"name":"Center for Stem Cell Research and Development (PEDI-STEM), Hacettepe University, 06100 Ankara, T\u00fcrkiye"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Cansu","family":"Ozdemir","sequence":"additional","affiliation":[{"name":"Department of Stem Cell Sciences, Institute of Health Sciences, Hacettepe University, 06100 Ankara, T\u00fcrkiye"},{"name":"Center for Stem Cell Research and Development (PEDI-STEM), Hacettepe University, 06100 Ankara, T\u00fcrkiye"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Gulen Eda","family":"Utine","sequence":"additional","affiliation":[{"name":"Department of Pediatric Genetics, Faculty of Medicine, Hacettepe University, 06100 Ankara, T\u00fcrkiye"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Pelin Ozlem","family":"Simsek-Kiper","sequence":"additional","affiliation":[{"name":"Department of Pediatric Genetics, Faculty of Medicine, Hacettepe University, 06100 Ankara, T\u00fcrkiye"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6040-6625","authenticated-orcid":false,"given":"Ekim Z.","family":"Taskiran","sequence":"additional","affiliation":[{"name":"Department of Medical Genetics, Faculty of Medicine, Hacettepe University, 06100 Ankara, T\u00fcrkiye"},{"name":"Department of Medical and Surgical Research, Institute of Health Sciences, Hacettepe University, 06100 Ankara, T\u00fcrkiye"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2025,1,28]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"319","DOI":"10.1016\/j.gene.2015.10.046","article-title":"Isolation, expression analysis and characterization of NEFA-interacting nuclear protein 30 and RING finger and SPRY domain containing 1 in skeletal muscle","volume":"576","author":"Waddell","year":"2016","journal-title":"Gene"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"187","DOI":"10.1002\/pro.3978","article-title":"The BioGRID database: A comprehensive biomedical resource of curated protein, genetic, and chemical interactions","volume":"30","author":"Oughtred","year":"2021","journal-title":"Protein Sci."},{"key":"ref_3","first-page":"D609","article-title":"UniProt: The Universal Protein Knowledgebase in 2025","volume":"53","author":"UniProt","year":"2024","journal-title":"Nucleic Acids Res."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"608","DOI":"10.1016\/j.ajhg.2015.08.007","article-title":"Identification of a Recognizable Progressive Skeletal Dysplasia Caused by RSPRY1 Mutations","volume":"97","author":"Faden","year":"2015","journal-title":"Am. J. Hum. Genet."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"2009","DOI":"10.1002\/ajmg.a.40427","article-title":"Further delineation of spondyloepimetaphyseal dysplasia Faden-Alkuraya type: A RSPRY1-associated spondylo-epi-metaphyseal dysplasia with cono-brachydactyly and craniosynostosis","volume":"176","author":"Taskiran","year":"2018","journal-title":"Am. J. Med. Genet. A"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"e63601","DOI":"10.1002\/ajmg.a.63601","article-title":"Two sisters with RSPRY1-related spondyloepimetaphyseal dysplasia","volume":"194","author":"Singh","year":"2024","journal-title":"Am. J. Med. Genet. A"},{"key":"ref_7","doi-asserted-by":"crossref","unstructured":"Lin, X., Patil, S., Gao, Y.G., and Qian, A. (2020). The Bone Extracellular Matrix in Bone Formation and Regeneration. Front. Pharmacol., 11.","DOI":"10.3389\/fphar.2020.00757"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"4195","DOI":"10.1242\/jcs.023820","article-title":"The extracellular matrix at a glance","volume":"123","author":"Frantz","year":"2010","journal-title":"J. Cell Sci."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"198","DOI":"10.1124\/pr.109.001289","article-title":"Extracellular matrix molecules: Potential targets in pharmacotherapy","volume":"61","author":"Jarvelainen","year":"2009","journal-title":"Pharmacol. Rev."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"1164","DOI":"10.1002\/ajmg.a.63132","article-title":"Nosology of genetic skeletal disorders: 2023 revision","volume":"191","author":"Unger","year":"2023","journal-title":"Am. J. Med. Genet A"},{"key":"ref_11","doi-asserted-by":"crossref","unstructured":"MacFarlane, E.G., Haupt, J., Dietz, H.C., and Shore, E.M. (2017). TGF-beta Family Signaling in Connective Tissue and Skeletal Diseases. Cold Spring Harb. Perspect. Biol., 9.","DOI":"10.1101\/cshperspect.a022269"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"101","DOI":"10.1038\/s41422-023-00918-9","article-title":"The roles and regulatory mechanisms of TGF-beta and BMP signaling in bone and cartilage development, homeostasis and disease","volume":"34","author":"Wu","year":"2024","journal-title":"Cell Res."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"211","DOI":"10.1046\/j.1523-1747.2002.01641.x","article-title":"Transforming growth factor-\u03b2 signaling through the Smad pathway:: Role in extracellular matrix gene expression and regulation","volume":"118","author":"Verrecchia","year":"2002","journal-title":"J. Investig. Dermatol."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"225","DOI":"10.1146\/annurev-genom-120922-094107","article-title":"TGF-beta and BMP Signaling Pathways in Skeletal Dysplasia with Short and Tall Stature","volume":"24","author":"Costantini","year":"2023","journal-title":"Annu. Rev. Genom. Hum. Genet."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"4007","DOI":"10.1016\/j.cell.2023.07.036","article-title":"TGF-j3 signaling in health and disease","volume":"186","author":"Sheppard","year":"2023","journal-title":"Cell"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"100245","DOI":"10.1016\/j.bonr.2020.100245","article-title":"Spondylo-epi-metaphyseal dysplasia due to a homozygous missense mutation in the gene encoding Matrilin-3 (T120M)","volume":"12","author":"Das","year":"2020","journal-title":"Bone Rep."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"26","DOI":"10.1097\/MCD.0000000000000241","article-title":"The second report on spondyloepimetaphyseal dysplasia, aggrecan type: A milder phenotype than originally reported","volume":"28","author":"Fukuhara","year":"2019","journal-title":"Clin. Dysmorphol."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"2832","DOI":"10.1172\/JCI22900","article-title":"mutation causes spondyloepimetaphyseal dysplasia, Missouri type (SEMD)","volume":"115","author":"Kennedy","year":"2005","journal-title":"J. Clin. Investig."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"60","DOI":"10.1038\/s41413-024-00364-2","article-title":"A monoallelic variant in causes an autosomal dominant spondyloepimetaphyseal dysplasia with low bone mass","volume":"12","author":"Li","year":"2024","journal-title":"Bone Res."},{"key":"ref_20","doi-asserted-by":"crossref","unstructured":"Rypdal, K.B., Apte, S.S., and Lunde, I.G. (2024). Emerging roles for the ADAMTS-like family of matricellular proteins in cardiovascular disease through regulation of the extracellular microenvironment. Mol. Biol. Rep., 51.","DOI":"10.1007\/s11033-024-09255-5"},{"key":"ref_21","doi-asserted-by":"crossref","unstructured":"Chen, E.Y., Tan, C.M., Kou, Y., Duan, Q., Wang, Z., Meirelles, G.V., Clark, N.R., and Ma\u2019ayan, A. (2013). Enrichr: Interactive and collaborative HTML5 gene list enrichment analysis tool. BMC Bioinform., 14.","DOI":"10.1186\/1471-2105-14-128"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"1830","DOI":"10.1093\/bioinformatics\/btt285","article-title":"Pathview: An R\/Bioconductor package for pathway-based data integration and visualization","volume":"29","author":"Luo","year":"2013","journal-title":"Bioinformatics"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"284","DOI":"10.1089\/omi.2011.0118","article-title":"clusterProfiler: An R Package for Comparing Biological Themes Among Gene Clusters","volume":"16","author":"Yu","year":"2012","journal-title":"Omics"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"31","DOI":"10.1089\/crispr.2018.0047","article-title":"Single-Step, High-Efficiency CRISPR-Cas9 Genome Editing in Primary Human Disease-Derived Fibroblasts","volume":"2","author":"Martufi","year":"2019","journal-title":"CRISPR J."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"W171","DOI":"10.1093\/nar\/gkz365","article-title":"CHOPCHOP v3: Expanding the CRISPR web toolbox beyond genome editing","volume":"47","author":"Labun","year":"2019","journal-title":"Nucleic Acids Res."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"W242","DOI":"10.1093\/nar\/gky354","article-title":"CRISPOR: Intuitive guide selection for CRISPR\/Cas9 genome editing experiments and screens","volume":"46","author":"Concordet","year":"2018","journal-title":"Nucleic Acids Res."},{"key":"ref_27","doi-asserted-by":"crossref","unstructured":"Suarez-Arnedo, A., Torres Figueroa, F., Clavijo, C., Arbelaez, P., Cruz, J.C., and Munoz-Camargo, C. (2020). An image J plugin for the high throughput image analysis of in vitro scratch wound healing assays. PLoS ONE, 15.","DOI":"10.1101\/2020.04.20.050831"}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/26\/3\/1134\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,8]],"date-time":"2025-10-08T10:37:41Z","timestamp":1759919861000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/26\/3\/1134"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,1,28]]},"references-count":27,"journal-issue":{"issue":"3","published-online":{"date-parts":[[2025,2]]}},"alternative-id":["ijms26031134"],"URL":"https:\/\/doi.org\/10.3390\/ijms26031134","relation":{},"ISSN":["1422-0067"],"issn-type":[{"type":"electronic","value":"1422-0067"}],"subject":[],"published":{"date-parts":[[2025,1,28]]}}}