Shard Chloe, Jones Anya C, Fouladzadeh Anahita, Palethorpe Helen M, Francis Abbie, Boyle Yasmin, Ormsby Rebecca J, Dewdney Brittany, Yeow Yen, Mahajan Ishika, Barker Matthew, Kuznetsova Irina, Jones Matthew E, Patil Ashwini, Rezaeiravesh Sara, Ying Ng Zi, Poonnoose Santosh I, Bosco Anthony, Valvi Santosh, Forrest Alistair R R, Johns Terrance G, Gomez Guillermo A, Fletcher Emily V
Centre for Cancer Biology, South Australia Pathology and University of South Australia, Adelaide, South Australia, 5000, Australia.
Cancer Centre, The Kids Research Institute Australia, Perth, Western Australia, 6009, Australia.
Neuro Oncol. 2025 Jun 11. doi: 10.1093/neuonc/noaf143.
Ion channel activity underlying biological processes that drive high-grade gliomas (HGG) is largely unknown. We aimed to determine the networking of ion channel genes and validate their expression within HGG patient tumors, to identify ion channel-targeting drugs that would inhibit tumor-promoting processes.
We used weighted gene co-expression network analysis (WGCNA) of RNAseq data to identify ion channel gene hubs in diffuse midline glioma (DMG) and glioblastoma. Using scRNA-seq, spatial transcriptomics, and immunohistochemistry, we characterized the expression of identified hubs within patient tumors, validating their role by testing the efficacy of ion channel inhibitors alone or in combination with radiation and temozolomide on the growth and invasion of patient-derived glioblastoma explant organoids (GBOs).
Network analysis revealed a preserved HGG "neuronal regulation" module, containing the greatest number of ion channels, with its corresponding genes concentrated at the tumor's leading-edge. Hubs within this module included γ-Aminobutyric-acid type A receptor (GABAAR) genes GABRA1 (α1) and GABRG2 (γ2), which immunohistochemically colocalized with GABAergic synaptic markers at the leading-edge. GBOs failed to retain this synaptic architecture but expressed a glioblastoma hub GABRA5 (α5), a component of extrasynaptic GABAARs. S44819, an α5-GABAAR antagonist strongly inhibited GBO invasion, with GABA(A)-compound 1b, a partial antagonist of GABAARs, robustly inhibiting GBO proliferation and invasion. Moreover, combined with standard of care (SOC) regimens, the anti-invasive properties of both compounds were enhanced in GBOs.
Our co-expression network analysis identified key ion channels at the leading-edge in HGGs, which can be targeted by GABAAR-acting drugs to disrupt tumor progression.
驱动高级别胶质瘤（HGG）的生物学过程背后的离子通道活性在很大程度上尚不清楚。我们旨在确定离子通道基因的网络，并在HGG患者肿瘤中验证其表达，以识别能够抑制肿瘤促进过程的靶向离子通道的药物。
我们使用RNA测序数据的加权基因共表达网络分析（WGCNA）来识别弥漫性中线胶质瘤（DMG）和胶质母细胞瘤中的离子通道基因枢纽。通过单细胞RNA测序、空间转录组学和免疫组织化学，我们对患者肿瘤中已识别的枢纽的表达进行了表征，通过测试离子通道抑制剂单独或与放疗和替莫唑胺联合使用对患者来源的胶质母细胞瘤外植体类器官（GBO）生长和侵袭的疗效来验证它们的作用。
网络分析揭示了一个保留的HGG“神经元调节”模块，其中包含最多数量的离子通道，其相应基因集中在肿瘤的前沿。该模块中的枢纽包括γ-氨基丁酸A型受体（GABAAR）基因GABRA1（α1）和GABRG2（γ2），它们在免疫组织化学上与前沿的GABA能突触标记物共定位。GBO未能保留这种突触结构，但表达了胶质母细胞瘤枢纽GABRA5（α5），这是突触外GABAAR的一个组成部分。α5-GABAAR拮抗剂S44819强烈抑制GBO侵袭，GABA（A）-化合物1b作为GABAAR的部分拮抗剂，有力地抑制GBO增殖和侵袭。此外，与标准治疗（SOC）方案联合使用时，两种化合物在GBO中的抗侵袭特性均得到增强。
我们的共表达网络分析确定了HGG前沿的关键离子通道，作用于GABAAR的药物可以靶向这些通道来破坏肿瘤进展。
