胶质母细胞瘤（GB）是最常见和最具侵袭性的脑肿瘤，对当前疗法表现出内在抵抗力，导致临床结果不佳。癌症进展部分归因于驱动癌细胞生长的蛋白质翻译机制的失调。在这项研究中，我们展示了 GB 的翻译组景观作为宝贵的数据资源。使用核糖体分析和信使 RNA (mRNA) 测序对八个患者来源的 GB 球培养物 (GSC) 进行了分析。我们通过翻译组和转录组水平的差异表达分析研究了细胞系间的差异。在 30 和 60 分钟时评估放疗后的转化变化。非编码 RNA (ncRNA) 的翻译使用内部和公共质谱 (MS) 数据进行验证，而 RNA 表达则通过定量 PCR (qPCR) 进行确认。我们的研究结果表明，核糖体测序提供了比 MS 或转录分析更详细的信息。 GSC 之间的转录相似性与翻译相似性相关，与先前定义的亚型（如原神经亚型和间充质亚型）一致。此外，我们在编码和非编码 mRNA 区域中发现了广泛的开放阅读框类型，包括长非编码 RNA (lncRNA) 和正在进行主动翻译的假基因。 ncRNA 到肽的翻译由内部数据和外部 MS 数据独立证实。我们还观察到组蛋白（下调）和剪接因子（上调）的翻译调节发生在放射治疗的反应中。这些数据为全基因组蛋白质合成提供了新的见解，识别了正常和放射治疗条件下 GB 中的翻译调控基因和替代翻译起始位点，为 GB 研究提供了丰富的资源。需要对放疗后差异表达基因进行进一步的功能验证。了解 GB 中的翻译控制可以揭示机制见解并识别当前未知的生物标志物，最终增强这种侵袭性脑癌的诊断和治疗。© 2024 作者。约翰·威利出版的《分子肿瘤学》

Glioblastoma (GB), the most common and aggressive brain tumor, demonstrates intrinsic resistance to current therapies, resulting in poor clinical outcomes. Cancer progression can be partially attributed to the deregulation of protein translation mechanisms that drive cancer cell growth. In this study, we present the translatome landscape of GB as a valuable data resource. Eight patient-derived GB sphere cultures (GSCs) were analyzed using ribosome profiling and messenger RNA (mRNA) sequencing. We investigated inter-cell-line differences through differential expression analysis at both the translatome and transcriptome levels. Translational changes post-radiotherapy were assessed at 30 and 60 min. The translation of non-coding RNAs (ncRNAs) was validated using in-house and public mass spectrometry (MS) data, whereas RNA expression was confirmed by quantitative PCR (qPCR). Our findings demonstrate that ribosome sequencing provides more detailed information than MS or transcriptional analyses. Transcriptional similarities among GSCs correlate with translational similarities, aligning with previously defined subtypes such as proneural and mesenchymal. Additionally, we identified a broad spectrum of open reading frame types in both coding and non-coding mRNA regions, including long non-coding RNAs (lncRNAs) and pseudogenes undergoing active translation. Translation of ncRNAs into peptides was independently confirmed by in-house data and external MS data. We also observed that translational regulation of histones (downregulated) and splicing factors (upregulated) occurs in response to radiotherapy. These data offer new insights into genome-wide protein synthesis, identifying translationally regulated genes and alternative translation initiation sites in GB under normal and radiotherapeutic conditions, providing a rich resource for GB research. Further functional validation of differentially expressed genes after radiotherapy is needed. Understanding translational control in GB can reveal mechanistic insights and identify currently unknown biomarkers, ultimately enhancing the diagnosis and treatment of this aggressive brain cancer.© 2024 The Author(s). Molecular Oncology published by John Wiley & Sons Ltd on behalf of Federation of European Biochemical Societies.