Expression of genes with biomarker potential identified in skin from DSLD-affected horses increases with age.
Abstract: 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 BMP2, FOS 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 KRT83 and BMP-2 in hair follicles in DSLD cases, as well as abnormally high expression of FOS in the epidermis, especially in aging horses. Because statistically relevant specificity and sensitivity was documented only for FOS and BMP2, but not KRT83 we recommend the use of FOS and BMP2 panel to diagnose DSLD. We conclude that a panel of two markers from the studied group (BMP2 and FOS) 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.
Copyright: © 2023 Roberts et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Publication Date: 2023-07-14 PubMed ID: 37450486PubMed Central: PMC10348567DOI: 10.1371/journal.pone.0287740Google Scholar: Lookup
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Summary
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The research article discusses the discovery of a potential biomarker panel for early detection and diagnosis of the progressive equine disease, Degenerative Suspensory Ligament Desmitis (DSLD) in living horses, particularly in older ones.
Overview of Degenerative Suspensory Ligament Desmitis (DSLD)
- DSLD is a systemic disorder that negatively affects connective tissues in horses, leading to chronic pain and lameness. This distressing condition also impacts multiple body systems of horses, including tendons, sclerae, and aorta.
- Currently, diagnosis of DSLD is only confirmed by post-mortem histological examination of a tendon or suspensory ligament. The histology shows abnormal accumulations of substances known as proteoglycans in affected tissues.
- Unfortunately, in living horses, there’s no reliable method existing for diagnosing DSLD.
Discovery of Potential Biomarkers for DSLD Diagnosis
- Bone morphogenetic protein 2 (BMP2) was recently identified to be present in active lesions of DSLD, marking its potential involvement in the disorder.
- Furthermore, data from RNA sequencing (RNA-seq) revealed overexpression of numerous genes, particularly BMP2, FOS (a known oncogene), and various keratin genes, in skin biopsies-derived RNA from horses with DSLD.
- The study hypothesizes that some of these overexpressed genes could be applied as diagnostic biomarkers in a panel for DSLD.
Verification of Biomarker Panel
- The overexpression of potential biomarkers was verified using quantitative real-time PCR, Immunohistochemistry, and RNAscope in-situ hybridization (ISH) assays.
- The ISH assay proved to be more reliable and specific than immunohistochemistry for this particular study.
- The ISH assay confirmed a significant increase in two of the potential biomarkers; KRT83 (a keratin gene) and BMP-2 in hair follicles, and an unusually high expression of FOS in the epidermis- mainly in ageing horses.
- However, the specificity and sensitivity were statistically significant only for FOS and BMP2 – not for KRT83 – endorsing the use of a FOS and BMP2 panel in DSLD diagnosis.
Conclusion and Future Implications
- The research concludes that a panel consisting of two markers (BMP2 and FOS) can act as an additional diagnostic tool in living horses, particularly older ones.
- However, further research is required to confirm if this biomarker panel would be prospective enough to detect DSLD in horses as they age.
Cite This Article
APA
Roberts JH, Zhang J, David F, McLean A, Blumenshine K, Müller-Alander E, Halper J.
(2023).
Expression of genes with biomarker potential identified in skin from DSLD-affected horses increases with age.
PLoS One, 18(7), e0287740.
https://doi.org/10.1371/journal.pone.0287740 Publication
Researcher Affiliations
- Department of Pathology, College of Veterinary Medicine, University of Georgia, Athens, Georgia, United States of America.
- Department of Pathology, College of Veterinary Medicine, University of Georgia, Athens, Georgia, United States of America.
- Equine Care Group, Mazy, Gembloux, Belgium.
- Equine Veterinary Medical Center-A member of Qatar Foundation, Doha, Qatar.
- Department of Animal Science, College of Agricultural and Environmental Science, University of California at Davis, Davis, California, United States of America.
- Santa Barbara Equine Practice, Santa Barbara, California, United States of America.
- Pferdepraxis, Overath, Germany.
- Department of Pathology, College of Veterinary Medicine, University of Georgia, Athens, Georgia, United States of America.
- Basic Science Department, AU/UGA Medical Partnership, Athens, Georgia, United States of America.
MeSH Terms
- Animals
- Horses
- Ligaments / pathology
- Skin / pathology
- Arthritis / pathology
- Proteoglycans
- Horse Diseases / diagnosis
- Horse Diseases / genetics
- Horse Diseases / pathology
- Lameness, Animal / pathology
Conflict of Interest Statement
The authors have declared that no competing interests exists.
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Citations
This article has been cited 2 times.- Jafari H, Abebe BK, Cong L, Ahmed Z, Zhaofei W, Sun M, Muhatai G, Chuzhao L, Dang R. Review: Genomic insights into the adaptive traits and stress resistance in modern horses. Stress Biol 2026 Jan 12;6(1):5.
- Roberts JH, Zhang J, David F, McLean A, Blumenshine K, Müller-Alander E, Halper J. Correction: Expression of genes with biomarker potential identified in skin from DSLD-affected horses increases with age. PLoS One 2025;20(6):e0326448.
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