Duan Wei, Chen Yu, Shan Jinlu, Li Qian
Department of Oncology, Jingzhou Central Hospital, The Second Clinical Medical College, Yangtze University, Jingzhou, Hubei, 434020, China.
Department of Radiology, Mianyang Central Hospital, School of Medicine, University of Electronic Science and Technology of China, Mianyang, 621000, China.
Recent Pat Anticancer Drug Discov. 2024;19(5):635-651. doi: 10.2174/0115748928259095231010055507.
Osteosarcoma (OS) is a common primary malignant bone tumor that mainly occurs in children and adolescents. The use of IL-8 inhibitor compounds has been reported in patents, which can be used to treat and/or prevent osteosarcoma, but the pathogenesis of osteosarcoma remains to be investigated. At present, osteoblasts and osteoclasts play an important role in the occurrence and development of OS. However, the relationship between osteoblasts and osteoclasts in the specific participation mechanism and inflammatory response of OS patients has not been further studied.
The transcriptome, clinical data, and other data related to OS were downloaded from the GEO database to analyze them with 200 known inflammatory response genes. We set the screening conditions as p < 0.05 and | logFC| > 0.50, screened the differentially expressed genes (DEGs) related to OS, tested the correlation coefficient between the OS INF gene and clinical risk, and analyzed the survival prognosis. We further enriched and analyzed the DEGs and inflammatory response genes of OS with GO/KEGG to explore the potential biological function and signal pathway mechanism of OS inflammatory response genes. Moreover, the virtual screening of drug sensitivity of OS based on the FDA drug library was also carried out to explore potential therapeutic drugs targeted to regulate OS osteogenesis and osteoclast inflammation, and finally, the molecular dynamics simulation verification of OS core protein and potential drugs was carried out to explore the binding stability and mechanism between potential drugs and core protein.
Through differential analysis of GSE39058, GSE36001, GSE87624, and three other data sets closely related to OS osteoblasts and osteoclasts, we found that there was one upregulated gene (CADM1) and one down-regulated gene (PHF15) related to OS. In addition, GSEA enrichment analysis of the DEGs of OS showed that it was mainly involved in the progress of OS through biological functions, such as oxidative photosynthesis, acute junction, and epithelial-mesenchymal transition. The enrichment analysis of OS DEGs revealed that they mainly affect the occurrence and progress of OS by participating in the regulation of the actin skeleton, PI3K Akt signal pathway, complement and coagulation cascade. According to the expression of CSF3R in OS patients, a risk coefficient model and a diagnostic model were established. It was found that the more significant the difference in the CSF3R gene in OS patients, the greater the risk coefficient of disease (p < 0.05). The AUC under the curve of the CSF3R gene was greater than 0.65, which had a good diagnostic significance for OS. The above results showed that the prognosis risk gene CSF3R related to OS inflammation was closely related to the survival status of OS patients. Finally, through the virtual screening of the ZINC drug library and molecular dynamics simulation, it was found that the docking model formed by the core protein CSF3R and the compounds, Leucovorin and Methotrexate, were the most stable, which revealed that the compounds Leucovorin and Methotrexate might play a role in the treatment of OS by combining with the inflammatory response related factor CSF3R of OS.
CSF3R participates in the occurrence and development of OS bone destruction by regulating the inflammatory response of osteoblasts and osteoclasts and can affect the survival prognosis of OS patients.
骨肉瘤（OS）是一种常见的原发性恶性骨肿瘤，主要发生于儿童和青少年。专利中已报道了白细胞介素-8抑制剂化合物的用途，其可用于治疗和/或预防骨肉瘤，但骨肉瘤的发病机制仍有待研究。目前，成骨细胞和破骨细胞在骨肉瘤的发生发展中起重要作用。然而，成骨细胞与破骨细胞在骨肉瘤患者具体参与机制及炎症反应方面的关系尚未得到进一步研究。
从基因表达综合数据库（GEO数据库）下载与骨肉瘤相关的转录组、临床数据等数据，与200个已知炎症反应基因进行分析。设定筛选条件为p < 0.05且|logFC| > 0.50，筛选出与骨肉瘤相关的差异表达基因（DEGs），检测骨肉瘤炎症相关基因与临床风险之间的相关系数，并分析生存预后。进一步用基因本体论（GO）/京都基因与基因组百科全书（KEGG）对骨肉瘤的差异表达基因和炎症反应基因进行富集分析，以探索骨肉瘤炎症反应基因的潜在生物学功能和信号通路机制。此外，还基于美国食品药品监督管理局（FDA）药物库对骨肉瘤药物敏感性进行虚拟筛选，以探索靶向调节骨肉瘤成骨和破骨细胞炎症的潜在治疗药物，最后对骨肉瘤核心蛋白与潜在药物进行分子动力学模拟验证，以探索潜在药物与核心蛋白之间的结合稳定性及机制。
通过对GSE39058、GSE36001、GSE87624以及其他三个与骨肉瘤成骨细胞和破骨细胞密切相关的数据集进行差异分析发现，与骨肉瘤相关的上调基因有1个（细胞粘附分子1，CADM1），下调基因有1个（植物同源结构域蛋白15，PHF15）。此外，骨肉瘤差异表达基因的基因集富集分析（GSEA）表明，其主要通过氧化光合作用、急性连接和上皮-间质转化等生物学功能参与骨肉瘤的进展。骨肉瘤差异表达基因的富集分析显示，它们主要通过参与肌动蛋白骨架、磷脂酰肌醇-3-激酶（PI3K）-蛋白激酶B（Akt）信号通路、补体和凝血级联反应的调控来影响骨肉瘤的发生和进展。根据骨肉瘤患者中集落刺激因子3受体（CSF3R）的表达情况，建立了风险系数模型和诊断模型。发现骨肉瘤患者中CSF3R基因差异越显著，疾病风险系数越大（p < 0.05）。CSF3R基因的曲线下面积（AUC）大于0.65，对骨肉瘤具有良好的诊断意义。上述结果表明，与骨肉瘤炎症相关的预后风险基因CSF3R与骨肉瘤患者的生存状况密切相关。最后，通过ZINC药物库虚拟筛选和分子动力学模拟发现，核心蛋白CSF3R与化合物亚叶酸钙和甲氨蝶呤形成的对接模型最稳定，这表明亚叶酸钙和甲氨蝶呤这两种化合物可能通过与骨肉瘤炎症反应相关因子CSF3R结合而在骨肉瘤治疗中发挥作用。
CSF3R通过调节成骨细胞和破骨细胞的炎症反应参与骨肉瘤骨破坏的发生发展，并可影响骨肉瘤患者的生存预后。
