TMIC-37. GEMISTOCYTIC TUMOR CELLS AND IMMUNE REACTIVE MICROGLIA PROGRAMMED FOR GLIAL SCARRING CHARACTERIZE T CELL CONTAINMENT IN IDH-MUTANT ASTROCYTOMA

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Abstract Isocitrate dehydrogenase 1/2 mutant (IDHmt) astrocytoma is considered a T cell-deprived tumor, yet little is known regarding the phenotype that relates to T cell exclusion. To aid better understanding of this mechanism, we systematically characterized the IDHmt astrocytoma immune microenvironment in a large cohort by multiplex immunofluorescence (IF) (n=75), bulk RNA sequencing (n=158), single nucleus RNA sequencing (n=7) and spatial RNA and protein profiling (n=12). We integrated and analyzed all datasets to acquire a robust characterization of the microenvironment organization and validated our results using IF stainings. We demonstrated that in IDHmt astrocytomas, T cells were contained in the perivascular space (i.e., T cell cuffs). In the tumor stroma surrounding T cell cuffs, we found a dense accumulation of gemistocytic tumor cells (GTCs), a histologically distinct tumor cell subtype. GTC-high tumors demonstrated enhanced expression of immune cell migration and activation pathways that were reliably expressed by TAMs and T cells. Notably, we found that GTCs formed a unique transcriptional cell sub-cluster that expressed markers including CD44, TNC and CRYAB, which are specific for glial scarring, a central nervous system specific mechanism for immune exclusion. Surrounding T cell cuffs, GTCs co-localized with immune-reactive microglia that expressed the chemoattractants CCL3 and CCL4. Together, co-localizing GTCs and reactive microglia expressed receptor-ligand pairs that take part in reactive astrogliosis and glial scarring, including Osteopontin – CD44 and IL-1β – IL1R1. Validation whole-slide multiplex IF stainings for this spatial relationship showed specific local accumulation of CD44 positive GTCs and TAMs around T cell cuffs in GTC-high tumors, which was absent in GTC-low tumors. Together, we show that IDHmt astrocytomas organize a network of tumor cells and leukocytes that mimics glial scarring and can aid T cell exclusion. Elucidating such specific immune exclusion mechanisms is vital for more appropriate target selection in novel immune therapies.