Abstract 1296: Integrated single cell spatial multi-omics landscape of WHO grades 2-4 diffuse gliomas identifies locoregional metabolomic correlates of glioma plasticity and proliferation.

Abstract Introduction: Diffuse infiltrating gliomas are aggressive tumors of the central nervous system driven by intra-tumoral heterogeneity and aberrant normal-tumor cell-cell interactions. Grade-specific and locoregional metabolic dependencies associated with aberrant transcriptional cell-states linked to treatment resistance and proliferation of gliomas remain elusive. We hypothesize that identifying metabolic surrogates of plasticity- and treatment-resistant glioma transcriptional cell states at the recurrence-prone tumor edge will lead to new therapeutic strategies for gliomas. Methods: We applied spatial transcriptomics (stRNAseq), imaging mass cytometry (IMC) and mass spectrometry imaging (MSI; metabolites, peptides and glycans) to the core and edge tumor tissue from patients with World Health Organization (WHO) grades 2-4 diffuse infiltrating gliomas including isocitrate dehydrogenase (IDH) mutant (mut) oligodendrogliomas (WHO Grades 2 and 3, n= 5 ) and IDH wildtype (wt) astrocytomas including ‘anaplastic’ astrocytoma (prior 2016 WHO histological grade 3, n = 3) and IDHwt glioblastoma (GBM, WHO grade 4). To determine if any differentially expressed genes between the core and edge of IDHwt GBM influenced overall survival (OS), we analyzed the association between OS and gene expression levels (low vs. high) in IDHwt GBM samples from the cancer genome atlas (TCGA). To identify grade specific and tumor edge unique metabolites that correlate with plasticity- and therapy-resistance-associated glioma transcriptional cell states and proliferation (Ki-67+), we integrated data from stRNAseq, IMC and MSI within each diffuse glioma grade. Results: High expression of Neuronatin (NNAT) and Heparin Sulfate-Glucosamine 3-Sulfotransferase 2 (HS3ST2) mRNA, which were significantly over-expressed in the IDHwt GBM edge relative to the core, were associated with significantly worse OS (NNAThi HR=1.8, p-value= 0.0043 and HS3ST2hi HR=2, p-value=0.0096). Integration of stRNA seq and MSI-derived metabolite expression identified edge unique metabolites overlapping with glioma transcriptional cell states and proliferative states across gliomas profiled. Conclusion: This comprehensive pan-diffuse infiltrating glioma multi-omics study could serve as a resource for not only uncovering region-specific metabolic signatures of transcriptionally defined cell states across WHO 2-4 diffuse gliomas, but also guide surgical extent of resection in real time, by using metabolic signatures as surrogates for plasticity associated transcriptional cell states and proliferation especially in the infiltrative and non-enhancing portion of the glioma edge. Citation Format: Yanxia Ma, Shamini Ayyadhury, Sanjay Singh, Yashu Vashishath, Cagri Ozdemir, Keziah Liebenberg, Trevor McKee, Nhat Nguyen, Akshay V. Basi, Duncan Mak, Javier A. Gomez, Jason T. Huse, Sana Noor, Sharon Wariko, Dan Winskowski, Regan Baird, Jeffrey S. Weinberg, Frederick F. Lang, Jared K. Burks, Livia S. Eberlin, Serdar Bozdag, Erin Seeley, Chibawanye I. Ene. Integrated single cell spatial multi-omics landscape of WHO grades 2-4 diffuse gliomas identifies locoregional metabolomic correlates of glioma plasticity and proliferation abstract. In: Proceedings of the American Association for Cancer Research Annual Meeting 2026; Part 1 (Regular Abstracts); 2026 Apr 17-22; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2026;86(7 Suppl):Abstract nr 1296.