Background:Osteosarcoma (OSA) is the most common type of bone cancer in canines. Novel therapies are required to prevent the growth, survival, and metastatic progression of this cancer, to increase life expectancy of patients. Immunohistochemical (IHC) studies and RNA sequencing help us gain a deeper understanding into the molecular mechanisms of the disease.Methods:We previously compared canine OSA tissues with patient matched non-tumour tissues, revealing 442 overexpressed genes within the samples. The present research used IHC staining for four of these genes in OSA tissues: G protein-coupled receptor 64 (GPR64), TOX High Mobility Group Box Family Member 3 (TOX3), Matrix Metallopeptidase 12 (MMP-12), and Forkhead Box F1 (FOXF1). H-scoring was performed to quantitatively assess protein expression and qualitatively contextualise staining locations. Additional analyses addressed whether gender or anatomical location of lesions (axial or appendicular tumours) affected protein expression. cBioPortal was employed to analyse expression and genetic alterations in patients.Results:GPR64,TOX3,MMP-12, andFOXF1showed high mRNA expression and genetic alterations in people with OSA. GPR64, TOX3, MMP-12, and FOXF1 were all expressed in canine OSA with novel findings regarding cellular expression. Additionally, differential sex expression was revealed for GPR64 and TOX3. Potential biomarkers or therapeutic targets were identified.Conclusions:These studies, and subsequent analysis, have provided insights into the molecular mechanisms associated with OSA progression and revealed potential biomarkers for diagnostic and prognostic purposes. A deeper understanding of genetic and protein interactions will support and progress novel pathways towards diagnostic, prognostic, and treatment interventions for OSA in both veterinary and human medicine.
背景：骨肉瘤（OSA）是犬类中最常见的骨癌类型。需要新的治疗方法来抑制该肿瘤的生长、存活及转移进程，以延长患者的预期寿命。免疫组化（IHC）研究和RNA测序有助于我们更深入地理解该疾病的分子机制。
方法：我们先前比较了犬骨肉瘤组织与患者匹配的非肿瘤组织，在样本中揭示了442个过表达基因。本研究通过免疫组化染色检测了骨肉瘤组织中其中四个基因的表达：G蛋白偶联受体64（GPR64）、TOX高迁移率族框家族成员3（TOX3）、基质金属肽酶12（MMP-12）以及叉头框F1（FOXF1）。采用H评分对蛋白表达进行定量评估，并对染色定位进行定性分析。进一步分析了性别或病变解剖位置（中轴骨或四肢骨肿瘤）是否影响蛋白表达。利用cBioPortal平台分析患者的基因表达及遗传变异情况。
结果：在人类骨肉瘤患者中，GPR64、TOX3、MMP-12和FOXF1显示出较高的mRNA表达水平和基因变异。在犬骨肉瘤组织中，GPR64、TOX3、MMP-12和FOXF1均有表达，并在细胞表达层面有新发现。此外，GPR64和TOX3的表达存在性别差异。研究识别了潜在的生物标志物或治疗靶点。
结论：这些研究及后续分析深入揭示了与骨肉瘤进展相关的分子机制，并发现了可用于诊断及预后评估的潜在生物标志物。对基因与蛋白相互作用的进一步理解，将推动兽医和人类医学中骨肉瘤诊断、预后及治疗干预新途径的发展。
Identification and Characterisation of Canine Osteosarcoma Biomarkers and Therapeutic Targets
肿瘤(癌症)患者之家