{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,3,6]],"date-time":"2026-03-06T01:12:14Z","timestamp":1772759534250,"version":"3.50.1"},"reference-count":40,"publisher":"Wiley","issue":"2","license":[{"start":{"date-parts":[[2022,2,7]],"date-time":"2022-02-07T00:00:00Z","timestamp":1644192000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc-nd\/4.0\/"}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["J cachexia sarcopenia muscle"],"published-print":{"date-parts":[[2022,4]]},"abstract":"Abstract<\/jats:title>\n \n Background<\/jats:title>\n The lack of dystrophin expression in Duchenne muscular dystrophy (DMD) induces muscle fibre and replacement by fibro\u2010adipose tissue. Although the role of some growth factors in the process of fibrogenesis has been studied, pathways activated by PDGF\u2010AA have not been described so far. Our aim was to study the molecular role of PDGF\u2010AA in the fibrotic process of DMD.<\/jats:p>\n <\/jats:sec>\n \n Methods<\/jats:title>\n \n Skeletal muscle fibro\u2010adipogenic progenitor cells (FAPs) from three DMD treated with PDGF\u2010AA at 50\u00a0ng\/mL were analysed by quantitative mass spectrometry\u2010based proteomics. Western\u2010blot, immunofluorescence, and G\u2010LISA were used to confirm the mass spectrometry results. We evaluated the effects of PDGF\u2010AA on the activation of RhoA pathway using two inhibitors, C3\u2010exoenzyme and fasudil. Cell proliferation and migration were determined by BrdU and migration assay. Actin reorganization and collagen synthesis were measured by phalloidin staining and Sircol assay, respectively. In an\n in vivo<\/jats:italic>\n proof of concept study, we treated dba\/2J\u2010mdx mice with fasudil for 6\u00a0weeks. Muscle strength was assessed with the grip strength. Immunofluorescence and flow cytometry analyses were used to study fibrotic and inflammatory markers in muscle tissue.\n <\/jats:p>\n <\/jats:sec>\n \n Results<\/jats:title>\n \n Mass spectrometry revealed that RhoA pathway proteins were up\u2010regulated in treated compared with non\u2010treated DMD FAPs (\n n<\/jats:italic>\n \u00a0=\u00a03, mean age\u00a0=\u00a08\u00a0\u00b1\u00a01.15\u00a0years old). Validation of proteomic data showed that Arhgef2 expression was significantly increased in DMD muscles compared with healthy controls by a 7.7\u2010fold increase (\n n<\/jats:italic>\n \u00a0=\u00a02, mean age\u00a0=\u00a08\u00a0\u00b1\u00a01.14\u00a0years old).\n In vitro<\/jats:italic>\n studies showed that RhoA\/ROCK2 pathway was significantly activated by PDGF\u2010AA (\n n<\/jats:italic>\n \u00a0=\u00a03, 1.88\u2010fold increase,\n P<\/jats:italic>\n \u00a0<\u00a00.01) and both C3\u2010exoenzyme and fasudil blocked that activation (\n n<\/jats:italic>\n \u00a0=\u00a03,\n P<\/jats:italic>\n \u00a0<\u00a00.05 and\n P<\/jats:italic>\n \u00a0<\u00a00.001, respectively). The activation of RhoA pathway by PDGF\u2010AA promoted a significant increase in proliferation and migration of FAPs (\n n<\/jats:italic>\n \u00a0=\u00a03,\n P<\/jats:italic>\n \u00a0<\u00a00.001), while C3\u2010exoenzyme and fasudil inhibited FAPs proliferation at 72\u00a0h and migration at 48 and 72\u00a0h (\n n<\/jats:italic>\n \u00a0=\u00a03,\n P<\/jats:italic>\n \u00a0<\u00a00.001).\n In vivo<\/jats:italic>\n studies showed that fasudil improved muscle function (\n n<\/jats:italic>\n \u00a0=\u00a05 non\u2010treated dba\/2J\u2010mdx and\n n<\/jats:italic>\n \u00a0=\u00a06 treated dba\/2J\u2010mdx, 1.76\u2010fold increase,\n P<\/jats:italic>\n \u00a0<\u00a00.013), and histological studies demonstrated a 23% reduction of collagen\u2010I expression area (\n n<\/jats:italic>\n \u00a0=\u00a05 non\u2010treated dba\/2J\u2010mdx and\n n<\/jats:italic>\n \u00a0=\u00a06 treated dba\/2J\u2010mdx,\n P<\/jats:italic>\n \u00a0<\u00a00.01).\n <\/jats:p>\n <\/jats:sec>\n \n Conclusions<\/jats:title>\n Our results suggest that PDGF\u2010AA promotes the activation of RhoA pathway in FAPs from DMD patients. This pathway could be involved in FAPs activation promoting its proliferation, migration, and actin reorganization, which represents the beginning of the fibrotic process. The inhibition of RhoA pathway could be considered as a potential therapeutic target for muscle fibrosis in patients with muscular dystrophies.<\/jats:p>\n <\/jats:sec>","DOI":"10.1002\/jcsm.12923","type":"journal-article","created":{"date-parts":[[2022,2,7]],"date-time":"2022-02-07T22:40:21Z","timestamp":1644273621000},"page":"1373-1384","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":23,"title":["RhoA\/ROCK2 signalling is enhanced by PDGF\u2010AA in fibro\u2010adipogenic progenitor cells: implications for Duchenne muscular dystrophy"],"prefix":"10.1002","volume":"13","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-4804-6553","authenticated-orcid":false,"given":"Esther","family":"Fern\u00e1ndez\u2010Sim\u00f3n","sequence":"first","affiliation":[{"name":"Neuromuscular Diseases Unit, Neurology Department Hospital de la Santa Creu i Sant Pau and Biomedical Research Institute Sant Pau (IIB Sant Pau) Barcelona 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