Liu Chaxian, Liu Yue, Lin Hao, Zhang Chufan, Zhang Bilong, Song Haikun, Fan Xiaomin, Lyu Yi, Yang Hui, Mao Ying
Department of Neurosurgery, Huashan Hospital, Fudan University, Shanghai, China.
Institute for Translational Brain Research, Shanghai Medical College, Fudan University, Shanghai, China.
Front Immunol. 2025 Apr 16;16:1587009. doi: 10.3389/fimmu.2025.1587009. eCollection 2025.
H3K27-altered diffuse midline glioma (DMG) is a highly aggressive glioma subtype, accounting for approximately 60% of pediatric high-grade gliomas, with a median survival of less than 12 months. Due to its predominant localization in the brainstem, conventional surgical resection is often unfeasible, underscoring the urgent need for alternative therapeutic strategies. While previous studies on DMG have primarily focused on regulatory mechanisms at the protein level, the role of alternative splicing in DMG remains largely unexplored. Given its potential impact on gene regulation and tumor progression, a comprehensive analysis of alternative splicing could provide novel insights into targeted or immune therapeutic strategies, complementing existing transcriptomic studies of DMG.
To investigate the alternative splicing landscape of DMG, we performed transcriptome sequencing (RNA-seq) on patient-derived H3WT and H3K27-altered DMG cell lines, integrating these data with RNA-seq and single-cell transcriptomic (scRNA-seq) datasets from published sources. This comprehensive approach enabled us to delineate the alternative splicing landscape of H3K27-altered DMG and validate its distinct features at the cellular level.
Our multi-omics analysis revealed significant transcriptional alterations in H3K27-altered DMG compared to H3WT DMG, particularly in pathways related to neuro-regulation, metabolism, and immunity. Further in-depth analysis identified extensive alternative splicing changes in H3K27-altered DMG, predominantly associated with RNA modifications and key alterations in extracellular matrix and nucleotide metabolism. Integrating these findings, we characterized five RNA-associated proteins that enabled a binary classification of DMG into neural and immune subtypes, with each subtype exhibiting distinct prognostic and transcriptomic features. Notably, we identified as a potential key regulator in DMG progression.
Our findings indicate that H3K27-altered DMG exhibits significant alternative splicing alterations, which play crucial roles in tumorigenesis and progression. Additionally, our study identified an RNA-binding protein-based classification of DMG and characterized as a potential regulatory factor, highlighting its potential as a novel therapeutic target.
H3K27改变的弥漫性中线胶质瘤（DMG）是一种高度侵袭性的胶质瘤亚型，约占儿童高级别胶质瘤的60%，中位生存期不到12个月。由于其主要位于脑干，传统手术切除往往不可行，这凸显了对替代治疗策略的迫切需求。虽然先前关于DMG的研究主要集中在蛋白质水平的调控机制，但可变剪接在DMG中的作用仍 largely未被探索。鉴于其对基因调控和肿瘤进展的潜在影响，对可变剪接的全面分析可为靶向或免疫治疗策略提供新的见解，补充现有的DMG转录组学研究。
为了研究DMG的可变剪接情况，我们对患者来源的H3WT和H3K27改变的DMG细胞系进行了转录组测序（RNA-seq），并将这些数据与已发表来源的RNA-seq和单细胞转录组学（scRNA-seq）数据集整合。这种综合方法使我们能够描绘H3K27改变的DMG的可变剪接情况，并在细胞水平验证其独特特征。
我们的多组学分析显示，与H3WT DMG相比，H3K27改变的DMG存在显著的转录改变，特别是在与神经调节、代谢和免疫相关的途径中。进一步深入分析发现，H3K27改变的DMG中存在广泛的可变剪接变化，主要与RNA修饰以及细胞外基质和核苷酸代谢的关键改变有关。综合这些发现，我们鉴定了五种与RNA相关的蛋白质，它们能够将DMG分为神经和免疫亚型，每个亚型具有不同的预后和转录组特征。值得注意的是，我们鉴定出 作为DMG进展中的一个潜在关键调节因子。
我们的研究结果表明，H3K27改变的DMG表现出显著的可变剪接改变，这些改变在肿瘤发生和进展中起关键作用。此外，我们的研究确定了基于RNA结合蛋白的DMG分类，并将 鉴定为一个潜在的调节因子，突出了其作为新型治疗靶点的潜力。
