A Functional Single-Nucleotide Polymorphism Upstream of the Collagen Type III Gene Is Associated with Catastrophic Fracture Risk in Thoroughbred Horses

Palomino Lago, Esther; Baird, Arabella; Blott, Sarah C.; McPhail, Rhona E.; Ross, Amy C.; Durward-Akhurst, Sian A.; Guest, Deborah J.

Abstract

Fractures caused by bone overloading are a leading cause of euthanasia in Thoroughbred racehorses. The risk of fatal fracture has been shown to be influenced by both environmental and genetic factors but, to date, no specific genetic mechanisms underpinning fractures have been identified. In this study, we utilised a genome-wide polygenic risk score to establish an in vitro cell system to study bone gene regulation in horses at high and low genetic risk of fracture. Candidate gene expression analysis revealed differential expression of COL3A1 and STAT1 genes in osteoblasts derived from high- and low-risk horses. Whole-genome sequencing of two fracture cases and two control horses revealed a single-nucleotide polymorphism (SNP) upstream of COL3A1 that was confirmed in a larger cohort to be significantly associated with fractures. Bioinformatics tools predicted that this SNP may impact the binding of the transcription factor SOX11. Gene modulation demonstrated SOX11 is upstream of COL3A1, and the region binds to nuclear proteins. Furthermore, luciferase assays demonstrated that the region containing the SNP has promoter activity. However, the specific effect of the SNP depends on the broader genetic background of the cells and suggests other factors may also be involved in regulating COL3A1 expression. In conclusion, we have identified a novel SNP that is significantly associated with fracture risk and provide new insights into the regulation of the COL3A1 gene.

Citation

Palomino Lago, E., Baird, A., Blott, S. C., McPhail, R. E., Ross, A. C., Durward-Akhurst, S. A., & Guest, D. J. (2024). A Functional Single-Nucleotide Polymorphism Upstream of the Collagen Type III Gene Is Associated with Catastrophic Fracture Risk in Thoroughbred Horses. Animals, 14(1), Article 116. https://doi.org/10.3390/ani14010116

• Journal Article Type: Article
• Acceptance Date: Dec 25, 2023
• Online Publication Date: Dec 28, 2023
• Publication Date: Jan 1, 2024
• Deposit Date: Jan 4, 2024
• Publicly Available Date: Jan 5, 2024
• Journal: Animals
• Electronic ISSN: 2076-2615
• Publisher: MDPI
• Peer Reviewed: Peer Reviewed
• Volume: 14
• Issue: 1
• Article Number: 116
• DOI: https://doi.org/10.3390/ani14010116
• Keywords: fracture risk; horse; genetic variation; bone; osteoblast; in vitro model
• Public URL: https://nottingham-repository.worktribe.com/output/29001519
• Publisher URL: https://www.mdpi.com/2076-2615/14/1/116