Next-Generation Sequencing in Translational Sarcoma Research: Biological Insights and Emerging Directions

Sarcomas are rare and heterogeneous malignancies for which molecular diagnostics increasingly complement histopathology, immunohistochemistry, FISH, and RT-PCR. This review summarizes the main research applications of next-generation sequencing (NGS) in sarcoma, with particular attention to mutation and variant discovery, fusion detection, biomarker development, assay validation, multi-omics integration, and the boundaries between exploratory research and clinical implementation.

NGS-based approaches have expanded the detection of recurrent and rare alterations across sarcoma subtypes, including subtype-defining fusions, copy-number changes, actionable kinase rearrangements, and alterations in tumor suppressor and cell-cycle pathways. However, the clinical utility of NGS remains context-dependent and should be interpreted according to tumor subtype, sample quality, available orthogonal methods, multidisciplinary expertise, and current precision-oncology recommendations. NGS is a powerful tool in translational sarcoma research and, in selected clinical scenarios, supports diagnosis, molecular classification, and therapeutic decision-making. Its use should be presented with appropriate caution, acknowledging technical limitations, the need for expert reference-center interpretation, and the complementary role of established diagnostic techniques.