While the poor prognosis of glioblastoma arises from the invasion of a subset of tumor cells, little is known of the metabolic alterations within these cells that fuel invasion. We integrated spatially addressable hydrogel biomaterial platforms, patient site–directed biopsies, and multiomics analyses to define metabolic drivers of invasive glioblastoma cells. Metabolomics and lipidomics revealed elevations in the redox buffers cystathionine, hexosylceramides, and glucosyl ceramides in the invasive front of both hydrogel-cultured tumors and patient site–directed biopsies, with immunofluorescence indicating elevated reactive oxygen species (ROS) markers in invasive cells. Transcriptomics confirmed upregulation of ROS-producing and response genes at the invasive front in both hydrogel models and patient tumors. Among oncologic ROS, H₂O₂ specifically promoted glioblastoma invasion in 3D hydrogel spheroid cultures. A CRISPR metabolic gene screen revealed cystathionine γ-lyase (CTH), which converts cystathionine to the nonessential amino acid cysteine in the transsulfuration pathway, to be essential for glioblastoma invasion. Correspondingly, supplementing CTH knockdown cells with exogenous cysteine rescued invasion. Pharmacologic CTH inhibition suppressed glioblastoma invasion, while CTH knockdown slowed glioblastoma invasion in vivo. Our studies highlight the importance of ROS metabolism in invasive glioblastoma cells and support further exploration of the transsulfuration pathway as a mechanistic and therapeutic target.

中文翻译：

---

侵袭性 GBM 细胞的多组学筛选揭示了可靶向的转硫途径改变

虽然胶质母细胞瘤的不良预后是由一部分肿瘤细胞的侵袭引起的，但人们对这些细胞内促进侵袭的代谢变化知之甚少。我们整合了空间可寻址水凝胶生物材料平台、患者定点活检和多组学分析，以确定侵袭性胶质母细胞瘤细胞的代谢驱动因素。代谢组学和脂质组学揭示了水凝胶培养肿瘤和患者定点活检的侵袭前沿中氧化还原缓冲液胱硫醚、己糖神经酰胺和葡萄糖神经酰胺的升高，免疫荧光表明侵袭细胞中的活性氧（ROS）标记物升高。转录组学证实了水凝胶模型和患者肿瘤中侵袭前沿的 ROS 产生和反应基因上调。在肿瘤学 ROS 中，H ₂ O ₂在 3D 水凝胶球体培养物中特异性促进胶质母细胞瘤侵袭。 CRISPR 代谢基因筛选显示，胱硫醚 γ-裂解酶 (CTH) 在转硫途径中将胱硫醚转化为非必需氨基酸半胱氨酸，对于胶质母细胞瘤的侵袭至关重要。相应地，用外源半胱氨酸补充 CTH 敲低细胞可挽救侵袭。药理学 CTH 抑制可抑制胶质母细胞瘤侵袭，而 CTH 敲除可减缓体内胶质母细胞瘤侵袭。我们的研究强调了活性氧代谢在侵袭性胶质母细胞瘤细胞中的重要性，并支持进一步探索转硫途径作为机制和治疗靶点。