Abstract
BACKGROUND: Chondrosarcoma, a rare and heterogeneous malignant bone tumor, presents significant clinical challenges due to its complex molecular underpinnings and limited treatment options. In this study, we employ single-cell RNA sequencing (scRNA-seq) and bioinformatics analyses to delineate cell subtypes, decipher signaling networks, and identify gene expression patterns, thereby providing novel insights into potential therapeutic targets and their implications in cancer biology. METHODS: scRNA-seq was performed on both clinical and experimental chondrosarcoma samples. Dimensionality reduction techniques (UMAP/t-SNE) were used to cluster cell subtypes, followed by Gene Ontology (GO) and pathway analyses to elucidate their biological functions. Cell-cell interaction networks, including the MIF signaling network, were reconstructed to map intercellular communications. Pseudotime analysis charted differentiation trajectories, while machine learning models evaluated the classification accuracy of gene expression patterns. GSEA was conducted to identify state-specific differential expression profiles. RESULTS: Over ten distinct cell subtypes were identified, including endothelial cells, fibroblasts, and epithelial cells. Key signaling pathways, such as TGF-beta signaling, focal adhesion, and actin cytoskeleton regulation, were found to mediate intercellular interactions. The MIF signaling network underscored the critical roles of immune cells within the tumor microenvironment. Pseudotime analysis revealed dynamic differentiation states, while state-specific gene expression patterns emerged from GSEA. Machine learning models demonstrated robust classification performance across training and external validation datasets. CONCLUSIONS: This comprehensive analysis uncovers the cellular heterogeneity and complex intercellular networks in chondrosarcoma, elucidating critical molecular pathways and identifying novel therapeutic targets. By integrating gene expression, signaling networks, and advanced computational methods, this study contributes to the broader understanding of cancer biology and highlights the potential for precision medicine strategies in treating chondrosarcoma.