Abstract
Background: Testicular seminomas, a common germ cell tumor, poses clinical challenges due to its molecular heterogeneity and limited biomarkers for precise diagnosis and prognosis. Leveraging multiomics approaches enables the comprehensive dissection of tumor complexity and facilitates the identification of key molecules influencing disease progression and therapeutic response. Methods: Single-cell RNA transcriptomic sequencing (scRNA-seq) was utilized to explore the cellular and transcriptional heterogeneity of testicular seminomas. High-dimensional weighted gene coexpression network analysis (hdWGCNA) identified gene modules linked to tumor progression. Public datasets were integrated for gene expression and survival analyses, and drug sensitivity patterns were assessed using the GDSC database. Results: scRNA-seq analysis revealed heterogeneous epithelial populations, with Epi1 cells exhibiting SLC5A5 and SPTBN4 as risk factors for advanced progression of seminomas. hdWGCNA identified nine gene modules, with the M6 module significantly enriched in Epi1 cells, implicating pathways such as negative regulation of ERAD and selective mRNA degradation. SPTBN4 was markedly upregulated in seminoma compared to nonseminomatous tumors and normal tissues, and its high expression was associated with poorer clinical outcomes and immunosuppressive microenvironments. Immune pathway analyses highlighted reduced antigen presentation and increased neutrophil extracellular trap (NET) formation in the SPTBN4-high group, suggesting diminished immunotherapeutic efficacy. Conversely, the SPTBN4-high group exhibited increased sensitivity to multiple chemotherapeutic agents, including thapsigargin and sorafenib, indicating its potential as a predictive marker for chemotherapy. Conclusion: In conclusion, this multiomics study identifies SPTBN4 as a central biomarker in testicular seminomas, encompassing diagnostic, prognostic, and therapeutic dimensions. The integration of single-cell transcriptomics, hdWGCNA, and drug sensitivity analyses underscores the molecular complexity of seminomas and highlights the translational potential of SPTBN4 in guiding personalized treatment strategies. These findings provide a foundation for leveraging multiomics approaches to advance the clinical management of testicular seminomas and other heterogeneous malignancies.