Zou Yunpeng, Kang Jianning, Zhu Shaopeng, Ren Xuechen, Li Zheng, Niu Jiayao, Qin Xuanzhe, Li Hongbo, Xiang Lu, Jiang Wei, Zhong Jiangbo, Zhang Ying, Zhao Kai
School of Clinical Medicine, Shandong Second Medical University, Weifang, Shandong, China.
Central Hospital Affiliated to Shandong First Medical University, Shandong First Medical University & Shandong Academy of Medical Sciences, Jinan, Shandong, China.
Front Immunol. 2025 Aug 15;16:1651858. doi: 10.3389/fimmu.2025.1651858. eCollection 2025.
Osteosarcoma (OS) is a malignant bone tumor with high metastatic potential and poor long-term survival. The tumor immune microenvironment and metabolic reprogramming are increasingly recognized as key drivers of OS progression, yet the molecular links between these systems remain unclear. This study aimed to identify immune-metabolic biomarkers in OS, focusing on pleckstrin (PLEK) as a potential regulatory hub.
We conducted differential expression and survival analyses using OS transcriptomic datasets and TCGA/GTEx data. Protein-protein interaction networks, GO/KEGG enrichment, and CytoHubba algorithms identified core hub genes. Tumor-infiltrating immune cells were profiled via TIMER. Single-cell RNA-seq (GSE162454) was used for immune and metabolic landscape mapping. PLEK was further validated by qRT-PCR and Western blot in OS samples, and its function assessed via siRNA knockdown in macrophages within TME co-cultured with OS cells. Cell proliferation, migration, and invasion assays evaluated phenotypic effects in OS cells.
Nine hub genes were identified, with PLEK significantly upregulated in OS tissues. High PLEK expression correlated with improved survival and increased infiltration of macrophages, dendritic cells, and CD4 T cells. Single-cell analysis showed PLEK enrichment in macrophage-dominated clusters with active glycolytic and oxidative phosphorylation pathways. Downregulation of PLEK in macrophages enhanced OS cell proliferation, migration and invasion. These findings suggest PLEK is linked to a pro-immune, metabolically active microenvironment and may act as a tumor suppressor.
Our study identifies PLEK as a prognostic biomarker and functional regulator in OS. It promotes an immune-infiltrated, metabolically active tumor microenvironment and is associated with attenuated malignant phenotypes . These findings highlight PLEK as a promising target for immunometabolic modulation in OS.
骨肉瘤（OS）是一种具有高转移潜能且长期生存率低的恶性骨肿瘤。肿瘤免疫微环境和代谢重编程日益被认为是骨肉瘤进展的关键驱动因素，但这些系统之间的分子联系仍不清楚。本研究旨在识别骨肉瘤中的免疫代谢生物标志物，重点关注普列克底物蛋白（PLEK）作为一个潜在的调控枢纽。
我们使用骨肉瘤转录组数据集以及TCGA/GTEx数据进行差异表达和生存分析。蛋白质-蛋白质相互作用网络、GO/KEGG富集分析以及CytoHubba算法确定了核心枢纽基因。通过TIMER对肿瘤浸润免疫细胞进行分析。利用单细胞RNA测序（GSE162454）绘制免疫和代谢图谱。通过qRT-PCR和蛋白质免疫印迹在骨肉瘤样本中进一步验证PLEK，并通过在与骨肉瘤细胞共培养的肿瘤微环境中的巨噬细胞中进行siRNA敲低来评估其功能。细胞增殖、迁移和侵袭实验评估了对骨肉瘤细胞表型的影响。
确定了9个枢纽基因，其中PLEK在骨肉瘤组织中显著上调。PLEK高表达与生存率提高以及巨噬细胞、树突状细胞和CD4 T细胞浸润增加相关。单细胞分析显示PLEK在以巨噬细胞为主的簇中富集，这些簇具有活跃的糖酵解和氧化磷酸化途径。巨噬细胞中PLEK的下调增强了骨肉瘤细胞的增殖、迁移和侵袭。这些发现表明PLEK与促免疫、代谢活跃的微环境相关，可能作为一种肿瘤抑制因子发挥作用。
我们的研究将PLEK鉴定为骨肉瘤中的一种预后生物标志物和功能调节因子。它促进免疫浸润、代谢活跃的肿瘤微环境，并与恶性表型减弱相关。这些发现突出了PLEK作为骨肉瘤免疫代谢调节的一个有前景的靶点。
