胶质母细胞瘤（GBM）是最具侵袭性和致命性的原发性脑肿瘤。传统治疗在改善生存率方面并没有取得突破。因此，需要寻找新的分子靶点和生物标志物。作为细胞膜上的信号传导坞站，四跨膜蛋白（TSPANs）与各种肿瘤相关；然而，关于它们在GBM中的作用的研究还极为有限。我们从GEPIA、CGGA、HPA、cBioPortal和GSCA数据库获取了基因表达和临床病理特征数据，分析了TSPANs在GBM中的mRNA和蛋白质表达水平、预后价值、临床相关性、突变状态和靶向药物敏感性。通过基因集富集分析（GSEA）、基因本体学（GO）和基因组学百科全书（KEGG）分析来进行生物学过程富集。TCGA和TCIA的数据用于构建TSPANs的肿瘤免疫微环境景观。采用不同的R软件算法分析免疫评分、免疫细胞浸润和免疫检查点相关性。对TSPAN4进行了单变量和多变量分析，它具有最显著的预测预后价值，并构建了一个预测个体结果的顺序图模型。在体内验证了TSPAN4的表达和功能。GBM中TSPAN3/4/6/11/12/18/23/24/25/26/27/28/29/30/31的表达显著上调，而TSPAN3/4/6/11/18/24/25/26/29/30与预后强相关。多个TSPAN的表达显著与1p/19q共缺失状态、IDH突变状态、复发、年龄和肿瘤分级相关。GSEA和GO分析揭示了TSPANs在细胞粘附和迁移中的潜在作用。免疫相关分析显示，TSPANs与GBM肿瘤微环境（TME）的形成有关，并可能影响免疫治疗结果。TSPAN4是一个独立的预后因子，TSPAN4敲除已被证明能够显著抑制体外胶质瘤细胞的增殖、侵袭和迁移。我们全面阐述了GBM中不同表达TSPANs的预后价值和潜在作用，其中包括科学家之前忽视的分子。该研究为GBM的病理机制和个体化肿瘤免疫治疗的未来方向提供了新颖而全面的视角，可能是GBM恶性进展与TME重塑之间的关键环节。© 2023. Springer Nature Limited.

Glioblastoma (GBM) is the most aggressive and lethal primary brain tumor. Conventional treatments have not achieved breakthroughs in improving survival. Therefore, novel molecular targets and biomarkers need to be identified. As signal transduction docks on the cell membrane, tetraspanins (TSPANs) are associated with various tumors; however, research on their role in GBM remains extremely scarce. Gene expression and clinicopathological characteristic data were obtained from GEPIA, CGGA, HPA, cBioPortal, and GSCA databases to analyze the mRNA and protein expression levels, prognostic value, clinical relevance, mutation status, and targeted drug sensitivity of TSPANs in GBM. Gene set enrichment analysis (GSEA), Gene Ontology (GO), and the Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis were used for biological process enrichment. Data from TCGA and TCIA were used to construct the tumor immune microenvironment landscape of TSPANs. Different R software algorithms were used to analyze the immune score, immune cell infiltration, and immune checkpoint correlation. Univariate and multivariate analyses were performed for TSPAN4, which had the most significant predictive prognostic value, and a nomogram model was constructed to predict individual outcomes. The expression and function of TSPAN4 were verified in vitro. TSPAN3/4/6/11/12/18/23/24/25/26/27/28/29/30/31expressions were significantly upregulated in GBM, and TSPAN3/4/6/11/18/24/25/26/29/30 were strongly correlated with prognosis. The expression of multiple TSPANs significantly correlated with 1p/19q co-deletion status, IDH mutation status, recurrence, age, and tumor grade. GSEA and GO analyses revealed the potential contribution of TSPANs in cell adhesion and migration. Immune correlation analysis revealed that TSPANs are related to the formation of the GBM tumor microenvironment (TME) and may influence immunotherapy outcomes. TSPAN4 is an independent prognostic factor and TSPAN4 knockdown has been demonstrated to strongly inhibit glioma cell proliferation, invasion, and migration in vitro. We comprehensively elaborated the prognostic value and potential role of differentially expressed TSPANs in GBM, including molecules that scientists have previously overlooked. This study provides a novel and comprehensive perspective on the pathological mechanisms of GBM and the future direction of individualized tumor immunotherapy, which may be a critical link between GBM malignant progression and TME remodeling.© 2023. Springer Nature Limited.