Data-Independent Acquisition-Based Mass Spectrometry (DIA-MS) Reveals Exosome Yes1 derived from cancer cells promotes osteosarcoma tumorigenesis via MAPK pathway

Abstract Background Osteosarcoma (OS) stands as the predominant primary bone tumor impacting children and young adults. Exosomes, vital for cellular communication, emerge as promising markers for diagnosing and prognosticating tumors. While few proteomic studies have delved into osteosarcoma's exosomal protein secretion, examining the differences in exosomal proteomes from OS cells can unveil insights into bone tumor development and spread. Methods This investigation employed Data-Independent Acquisition-Based Mass Spectrometry to analyze the exosomal proteomes of osteoblast and osteosarcoma cells. Exosomal YES1, belonging to the proto-oncogene tyrosine-protein kinase Src family kinases and linked with cancer genesis, tumor environments, and patient survival across various cancers, especially osteosarcoma, was a focal point. The study's experimental approach included in vitro functional assays (Edu, wound healing, transwell assays, flow cytometry) and in vivo experiments to assess the effects of exosomes on the malignant traits of OS cells. Immunohistochemistry was used to compare YES1 expression in human osteosarcoma tissues against normal tissues, complemented by bioinformatic analyses correlating YES1 expression levels with patient survival and prognosis. Results The research found that exosomal YES1 from osteosarcoma cells is integral to the immune microenvironment and cancer-promoting activities in OS through the MAPK pathway. It was observed that YES1 expression was notably higher in osteosarcoma tissues compared to normal ones, aligning with bioinformatics findings that linked elevated YES1 expression with lower overall survival rates and poorer prognoses in patients. Conclusions This study highlighted the critical role of osteosarcoma-derived exosomal YES1 in tumor immunology and oncogenesis via the MAPK pathway, presenting new perspectives on the function of exosomal proteins in osteosarcoma tumorigenesis. It also suggests the potential of exosomal YES1 as a biomarker for osteosarcoma therapy, emphasizing its importance in understanding tumor behavior and improving patient outcomes.