{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,1,12]],"date-time":"2026-01-12T06:11:32Z","timestamp":1768198292149,"version":"3.49.0"},"reference-count":16,"publisher":"Public Library of Science (PLoS)","issue":"7","license":[{"start":{"date-parts":[[2023,7,14]],"date-time":"2023-07-14T00:00:00Z","timestamp":1689292800000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"USDA Animal Health Capacity Grant"},{"name":"USDA Animal Health Capacity Grant"},{"name":"Global Research Collaborative Grant"},{"name":"Global Research Collaborative Grant"},{"name":"ORFGA Grant, Summer Research Stipends and Grants"},{"name":"Foundation for Horses and Other Animals"},{"name":"Mr. and Mrs. Lou Gonda"},{"name":"Mr. and Mrs. Lou Gonda"},{"name":"Mr. and Mrs. Lou Gonda"},{"name":"El Campeon Farm"},{"name":"El Campeon Farm"}],"content-domain":{"domain":["www.plosone.org"],"crossmark-restriction":false},"short-container-title":["PLoS ONE"],"abstract":"<jats:p>Degenerative Suspensory Ligament Desmitis (DSLD) negatively impacts connective tissues in horses, which often leads to progressive chronic pain and lameness. DSLD has been shown to be a systemic disorder that affects multiple body systems, including tendons, sclerae, and the aorta. Currently, the diagnosis is confirmed by post mortem histological examination of a tendon or suspensory ligament. Histology reveals inappropriate accumulations of proteoglycans in the tendons and other tissues in DSLD-affected horses. Unfortunately, there is no reliable method to diagnose DSLD in living horses. Recently, bone morphogenetic protein 2 (BMP2) was identified in active DSLD lesions. In addition, recent data from RNA sequencing (RNA-seq) showed overexpression of numerous genes, among them <jats:italic>BMP2<\/jats:italic>, <jats:italic>FOS<\/jats:italic> and genes for keratins in DSLD skin biopsies-derived RNA. We hypothesized that some of these genes can be used as biomarkers for diagnosis of DSLD in a panel. Overexpression of some of them was verified in quantitative real time PCR. Immunohistochemistry and RNAscope in-situ hybridization (ISH) assays were used to determine the level of overexpression of specific genes in skin biopsies from control and DSLD-affected horses. The RNAscope ISH assay has shown to be more reliable and more specific that immunohistochemistry. ISH confirmed a significant increase in <jats:italic>KRT83<\/jats:italic> and <jats:italic>BMP-2<\/jats:italic> in hair follicles in DSLD cases, as well as abnormally high expression of <jats:italic>FOS<\/jats:italic> in the epidermis, especially in aging horses. Because statistically relevant specificity and sensitivity was documented only for <jats:italic>FOS<\/jats:italic> and <jats:italic>BMP2<\/jats:italic>, but not <jats:italic>KRT83<\/jats:italic> we recommend the use of <jats:italic>FOS<\/jats:italic> and <jats:italic>BMP2<\/jats:italic> panel to diagnose DSLD. We conclude that a panel of two markers from the studied group (<jats:italic>BMP2<\/jats:italic> and <jats:italic>FOS<\/jats:italic>) can serve as an additional diagnostic tool for DSLD in living horses, especially in older animals. Further studies are necessary to confirm if this biomarker panel could be used as a prospective tool to identify DSLD in horses as they age.<\/jats:p>","DOI":"10.1371\/journal.pone.0287740","type":"journal-article","created":{"date-parts":[[2023,7,14]],"date-time":"2023-07-14T17:36:52Z","timestamp":1689356212000},"page":"e0287740","update-policy":"https:\/\/doi.org\/10.1371\/journal.pone.corrections_policy","source":"Crossref","is-referenced-by-count":2,"title":["Expression of genes with biomarker potential identified in skin from DSLD-affected horses increases with age"],"prefix":"10.1371","volume":"18","author":[{"given":"Jennifer Hope","family":"Roberts","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jian","family":"Zhang","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1933-2219","authenticated-orcid":true,"given":"Florent","family":"David","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Amy","family":"McLean","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Karen","family":"Blumenshine","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Eva","family":"M\u00fcller-Alander","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0233-0567","authenticated-orcid":true,"given":"Jaroslava","family":"Halper","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"340","published-online":{"date-parts":[[2023,7,14]]},"reference":[{"key":"pone.0287740.ref001","doi-asserted-by":"crossref","first-page":"12","DOI":"10.1186\/1746-6148-2-12","article-title":"Degenerative suspensory ligament desmitis as a systemic disorder characterized by proteoglycan accumulation","volume":"2","author":"J. 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