BackgroundOsteosarcoma is an aggressive and painful bone neoplasm in dogs. Previous studies have reported epidemiological associations suggesting that large body mass, long bone length and the genetics of certain breeds including the Rottweiler are associated with elevated osteosarcoma risk. However, these studies were often limited by selection bias and confounding, and have rarely offered insights into breed-associated protection as well as breed-associated predisposition for osteosarcoma. The current study includes 1756 osteosarcoma cases presenting to VPG Histology (Bristol, UK) that are compared against a control population of 905,211 dogs without osteosarcoma taken from primary care electronic patient records in the VetCompass™ dataset. Methods and Study DesignRetrospective, case-control study. Multivariable logistic regression analysis explored associations between demographic risk factors (including breed, chondrodystrophy, age, sex/neuter status, skull-shape, and body mass) and osteosarcoma. ResultsWe identified several breeds with increased and reduced odds of osteosarcoma. At highest risk were the Rottweiler and Great Dane, with >10 times the odds of osteosarcoma compared with crossbreeds, and the Rhodesian Ridgeback; which has not featured in previous lists of at-risk breeds for osteosarcoma, and had an odds ratio of 11.31 (95% confidence interval 7.37 - 17.35). Breeds at lowest risk of osteosarcoma (protected breeds) included the Bichon Frise, the French Bulldog and the Cavalier King Charles Spaniel, all with odd ratios of less than 0.30 compared with crossbreeds. Body mass was strongly associated with osteosarcoma risk; dogs over 40kg exhibited osteosarcoma odds of 45.44 (95% confidence interval 33.74 - 61.20) compared with dogs less than 10kg. Chondrodystrophic breeds had an osteosarcoma odds ratio of 0.13 (95% confidence interval 0.11 - 0.16) compared with non-chondrodystrophic breeds. ConclusionsOverall, this study provides evidence of strong breed-associated osteosarcoma risk and protection, suggesting a genetic basis for osteosarcoma pathogenesis. The study findings highlight that breeds bred for long legs or large body mass are generally overrepresented amongst at-risk breeds, whilst those bred for short leg length or small body mass are generally protected. These findings could inform genetic studies to identify osteosarcoma risk alleles in canines and humans; as well as increasing awareness amongst vets and owners, resulting in improved breeding practices and clinical management of osteosarcoma in dogs.