Abstract 1375: The intra-tumoral spatial heterogeneity of T-cell antigens in glioblastoma: An integrated multi-omics approach
Authors: Wacker Marcel , Medici Gioele , Dubbelaar Marissa , Bauer Jens , Nelde Annika , Regli Luca , Weller Michael , Salih Helmut R. , Rammensee Hans-Georg , Neidert Marian C. , Walz Juliane S. 
Published: June 2022

Abstract
Glioblastoma is the most common primary malignant neoplasm of the central nervous system in adults. Current treatment options comprise maximal surgical resection followed by radiation and/or chemotherapy with temozolomide. However, these procedures are unable to eliminate all tumor cells, which in turn lead to disease recurrence accounting for the poor prognosis. Glioblastoma is a highly infiltrative tumor in which recurrence originates from the unresectable peritumoral infiltration zone. Thus, novel treatment options specifically targeting the tumor cells in the infiltration zone are needed to prevent relapse and enable long-lasting remission. In this work, we performed multi-omics spatial analysis of the necrotic center (NEC), the gadolinium contrast-enhanced region (T1), and the infiltration zone (INF) to assess immunological relevant aspects of tumor heterogeneity. By integrating mass spectrometry-based immunopeptidome analysis with next generation sequencing methods (whole exome and RNA sequencing, DNA methylation), we aimed to identify the intra-tumoral regional heterogeneity of T cell antigens with a special focus on the peritumoral infiltration zone. For multi-omics analysis, HLA peptides and genetic material from 15 glioblastoma patients were extracted and analyzed from the three zones NEC, T1, INF and for four patients additionally from adjacent benign (BEN) brain tissue. A total of 24,699 unique HLA class I and 17,394 HLA class II peptides were identified. Comparative profiling of peptides from our study and a benign tissue database (in-house (n=429) combined with HLA ligand atlas (https://hla-ligand-atlas.org)) revealed that 6% (970/15,579), 5% (829/16,442) and 5% (725/13,891) of HLA class I peptides were exclusively presented in the INF, T1 and NEC zone, respectively. 6% (394/7,150), 7% (589/8,736) and 13% (1,725/12,908) of HLA class II peptides showed exclusive representation in the INF, T1 and NEC zone, respectively. Importantly, due to the spatial analysis of the immunopeptidome, we revealed 970 HLA class I and 394 HLA class II peptides exclusively presented in the INF zone. Interestingly, one INF-associated HLA class I peptide, which showed frequent presentation in 36% of glioblastoma immunopeptidomes, is derived from the brain and acute leukemia cytoplasmic protein (BAALC), a highly expressed cell cycle inducer (via MEK kinase-1) in several cancers, including glioblastoma. Integrated RNA/DNA sequencing enabled a greater understanding of spatial tumor antigen presentation and lead to the identification of INF-specific neoepitopes derived from tumor-specific mutations. In summary, we identified the intra-tumoral regional heterogeneity of tumor antigens, which could be used in the future for specific immunotherapy approaches targeting the infiltration zone of glioblastoma.
Citation Format: Marcel Wacker, Gioele Medici, Marissa Dubbelaar, Jens Bauer, Annika Nelde, Luca Regli, Michael Weller, Helmut R. Salih, Hans-Georg Rammensee, Marian C. Neidert, Juliane S. Walz. The intra-tumoral spatial heterogeneity of T-cell antigens in glioblastoma: An integrated multi-omics approach [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 1375.