IMMU-58. CHARACTERIZATION OF MYELOID CELL POPULATIONS IN PEDIATRIC HIGH-GRADE BRAIN TUMORS: THE MIMIC PROGRAM

Abstract

Abstract BACKGROUND Pediatric high-grade brain tumors (pHGBT) pose treatment challenges with limited options. Myeloid cells, particularly tumor-associated macrophages (TAMs) in the tumor-microenvironment (TME), exhibit potent immunosuppressive features inhibiting effective anti-tumor T-cell responses. Targeting these cells holds promise to improve outcomes, however the underlying mechanisms of myeloid cell dysfunction remain poorly understood. This study aimed to comprehensively characterize myeloid cell populations in pHGBT and investigate their functional phenotype. METHODS Spectral flow cytometry was performed on tumor tissue (biopsies or resections) and tumor fragments obtained by cavitron ultrasonic surgical aspiration (CUSA) in pHGBT patients (High-grade glioma n=2, Ependymoma n=1, Medulloblastoma n=6, Pilocytic Astrocytoma n=2) enrolled in the MIMIC program. This allowed quantification and comprehensive characterization of myeloid cell populations within the TME. RESULTS The pHGBT TME showed a low abundance of immune cells across different entities, with myeloid cells comprising the predominant proportion. CD49d was used to identify invading cells derived from bone marrow (CD49+) and resident microglia (CD49-). The total TAMs/microglia population was detected in both tumor tissue and CUSA tumor fragments in similar proportions, however resident CD49d- microglia were notably increased in CUSA tumor fragments. Additionally, invading TAMs displayed a mixed "M1" and "M2" polarization phenotype and exhibited higher expression of the chemokine receptor CCR2 and HLA-DR, and of the immune-checkpoint receptors SIRP1A/B, CD33, and TIM3, compared to resident microglia. CONCLUSIONS TAMs play a multifaceted role in pHGBT. The high expression of immune-checkpoint receptors by invading TAMs suggests an immunosuppressive phenotype, potentially facilitating tumor growth, infiltration and (immune)therapy escape. Although, differences between disease entities could not yet be analyzed, disparities were observed in the TME between tumor and CUSA samples, likely related to spatial cellular organization. Understanding TAM dynamics and functional differences between invading and resident subsets, as well as their underlying mechanisms, is crucial for advancing therapeutic efficacy in pHGBT.