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.
View: Text | PDF
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

×

Figure 3

Gene-expression profiling demonstrates upregulated pathways involved in adapting to oxidative stress in invasive GBM cells.

Options: View larger image (or click on image) Download as PowerPoint
Gene-expression profiling demonstrates upregulated pathways involved in ...
(A and B) RNA from invasive and core (A) GBM43 cells from hydrogel invasion devices or (B) site-directed biopsies of patient GBMs were assessed using the NanoString nCounter panel, which analyzes expression of 770 genes from 34 metabolic pathways, with GSEA revealing enriched metabolic pathways, including 5 shared between GBM43 cells in hydrogels and patient specimens (green). Volcano plots (P and FC = probability of significance and fold change invasive versus core) are shown for genes in 2 of these pathways — mitochondrial respiration (left) and ROS response genes (right) — highlighting genes in invasive (log2FC > 0) and core (log2FC < 0) samples. (C–F) Bulk RNA-Seq on invasive and core GBM43 cells isolated from hydrogels revealed the following: (C) Of 2,172 up- or downregulated (Padjusted < 0.05) genes in invasive versus core GBM43 cells (gray dots on volcano plot), 344 (16%) were involved in cellular metabolism (green dots = upregulated genes, pink dots = downregulated genes). (D) Among 2,172 up- or downregulated (Padjusted < 0.05) genes in invasive versus core GBM43 cells (gray dots on volcano plot), shown are the 10 most up- and downregulated metabolic genes (green dots = upregulated genes, pink dots = downregulated genes and listed accordingly in the graph to the right). (E) KEGG pathway analysis of genes enriched in invasive GBM cells implicated pathways involved in the production of and response to ROS. (F) Gene-expression changes overlaid on an oxidative phosphorylation schematic revealed upregulated genes encoding mitochondrial complexes II–V in invasive GBM43 cells versus those in the core.