Multiomic screening of invasive GBM cells reveals targetable transsulfuration pathway alterations
Joseph H. Garcia, … , Sanjay Kumar, Manish K. Aghi
Joseph H. Garcia, … , Sanjay Kumar, Manish K. Aghi
Published November 16, 2023
Citation Information: J Clin Invest. 2024;134(3):e170397. https://doi.org/10.1172/JCI170397.
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Research Article Metabolism Oncology

Multiomic screening of invasive GBM cells reveals targetable transsulfuration pathway alterations

Abstract

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.

Authors

Joseph H. Garcia, Erin A. Akins, Saket Jain, Kayla J. Wolf, Jason Zhang, Nikita Choudhary, Meeki Lad, Poojan Shukla, Jennifer Rios, Kyounghee Seo, Sabraj A. Gill, William H. Carson, Luis R. Carette, Allison C. Zheng, David R. Raleigh, Sanjay Kumar, Manish K. Aghi

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Figure 2

Lipidomic profiling indicates increased oxidative stress, lipid peroxidation, and apoptotic signaling at the invasive GBM front.

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Lipidomic profiling indicates increased oxidative stress, lipid peroxida...
Shown are results from unbiased lipidomic analysis of cells from the invasive front and tumor core of GBM43 cells in 3D hydrogels and site-directed biopsies of patient GBMs. (A) Volcano plots displaying relative fold change for individual lipid abundance at the invasive front of hydrogels (left) and patient specimens (right) versus tumor core. (B) Heatmaps displaying relative abundance of lipids in hydrogels (left) and patient specimens (right) organized by lipid classification. (C) Relative fold change of hexosylceramide and glucosylceramide species at the invasive tumor front in hydrogels (left) and patient tumors (right). Data are represented as mean ± SD. *P < 0.05, t test. (D) Illustration of pathways enabling hexosylceramide and glucosylceramide species to protect against apoptosis in invasive GBM cells exposed to oxidative stress. (E) KEGG pathway enrichment analysis of untargeted lipidomics displaying lipid pathways upregulated at the invasive tumor front of hydrogels (left) and patient tumors (right) using bubble plots.