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Identification of the Key Serum Biomarkers to Diagnose and Predict Metastasis of Osteosarcoma Based on the Analysis of Immune Cell Infiltration Characteristics


 Background: The potential functions of circular RNAs (circRNAs) and micro RNAs (miRNAs) in osteosarcoma (OS) have not been fully elucidated. Especially, the behavior and mechanism of immune responses in OS development and progression have not been fully demonstrated. It was reported that circRNAs and miRNAs can serve as biomarkers for the diagnosis, prognosis, and therapy of many cancers. This study aimed to identify novel key serum biomarkers to diagnose and predict metastasis of OS based on the analysis of immune cell infiltration characteristics.Methods: The differentially-expressed circRNAs (DEcircRNAs), differentially-expressed miRNAs (DEmiRNAs),and differentially-expressed mRNAs (DEmRNAs) of human OS were investigated based on the microarray data downloaded from Gene Expression Omnibus (GEO) datasets. Then, we analyzed immune characteristics pattern of tumor-infiltrating immune cells in OS. On this basis, we identified statistically-significant transcription factors and performed pathway enrichment analysis. Subsequently, we constructed protein-protein interaction (PPI) and competitive endogenous RNA (ceRNA) networks. Moreover, the biological characteristic of targets in ceRNA networks was proposed. Finally, the expression and diagnostic capability of these potential biomarkers from ceRNA network were confirmed by RT-qPCR in patients’ serum.Results: Seven differentially-expressed circRNAs (DEcircRNAs), 166 differentially-expressed miRNAs (DEmiRNAs) and 175 differentially-expressed mRNAs (DEmRNAs) were identified in total. The highest level of infiltration in OS patients were M0 macrophages, M2 macrophages and CD8+ T cells. Further, M0 macrophages and CD8+ T cells were showed the largest negative correlation coefficients. These significant immune characteristics pattern of tumor-infiltrating immune cells were revealed by the principal component analysis in OS. Moreover, we found 185 statistically-significant transcription factors in which the main significant molecules show the potential in immunotherapy of OS. Hsa-circ-0010220, hsa-miR-326, hsa-miR-338-3p, and FAM98A from ceRNA networks associated with immune cell infiltration were confirmed as the potential novel biomarkers for OS diagnosis, of which FAM98A could distinguish and predict metastasis. Most importantly, a novel diagnostic model consisting of the four promising biomarkers (hsa-circ-0010220, hsa-miR-326, hsa-miR-338-3p, and FAM98A) was highlighted with 0.928 AUC value.Conclusions: In summary, the potenial serum biomarkers to diagnose and predict metastasis of OS based on the analysis of immune cell infiltration characteristics were found, and a novel diagnostic model consisting of four promising serum biomarkers was proposed firstly. These results provided a new perspective for the immunotherapy of OS.