IMMU-82. Neuronal modulation of immunity in glioblastoma

Abstract:

Abstract Recent studies determined that in glioblastoma, tumor cells integrate into neural networks, forming synapses with neurons, enhancing neuronal firing facilitating tumor growth. We sought to understand how neuronal activity alters the function of immune cells in glioblastoma using both murine models and freshly acquired human glioblastoma tissue from surgical specimens. We first used chemogenetic stimulation (AAV5-SYN1-hM3Dq) of an RCAS-tva mouse glioblastoma model (Ntv-a Ink4a-Arf-/-) to identify which tumor immune populations are impacted by enhanced neuronal firing in vivo. Using high-dimensional spectral flow cytometry, we found an increase in microglia following stimulation. We then used human glioblastoma tissue to study a possible neuron-myeloid interaction. Immunofluorescent staining of these tumors showed that approximately 50% of the Iba1+ myeloid cells colocalized with the presynaptic neuronal marker synapsin 1, confirming the neuron-myeloid interaction. We then sorted CD11b+ myeloid cells from these patient samples. Bulk RNA-seq revealed high expression of neurotransmitter receptors, predominantly the Gi protein-coupled P2Y12 receptor for ADP. Calcium fluorescence imaging, calcium flow cytometry, and a phagocytic assay showed that the cells were primarily responsive to ADP among the neurotransmitters examined, including glutamate, norepinephrine, GABA, and acetylcholine. Analysis of bulk RNA-seq from 286 brain tumor patients found that P2Y12 had a higher positive correlation with M1-like markers and a negative correlation with M2-like markers than all other neurotransmitter receptor genes and others such as ADRB2, CX3CR1, TMEM119, TREM2, and CSF1R. To understand how the ADP-P2Y12 axis affects myeloid phenotypic function in vivo, we conducted flow cytometry on our SB28 mouse brain tumor tissues. We found that P2Y12-positive microglia had an increased M1-like and decreased M2-like phenotype compared to P2Y12-negative microglia, based on functional marker expression. Overall, we conclude that neuronal excitation affects microglia in the glioblastoma microenvironment, primarily through an ADP-P2Y12 axis, which could be an important target for enhancing response to immunotherapy.

Authors:

JC Hancock, Q Zhao, T Shaw, J Seaman, N Briceno, M Pavan, P Olivia, J Lee, H Naylor, M Pitoulias, V Pisupati, K Kishore, L Porcu, AY Mitrophanov, M Natoli, J Miller, M Santos, M Zhang, D Davis, W Zhang, H Song, J Spurgeon, H Kim, A Hara, C Huguely, B Anderson, E Steffke, S Keretsu, AT Chesler, C Yang, H Robinson, KA Jacobson, MR Gilbert, M Terabe, R Mair

Journal:

Neuro-Oncology

Citation info:

27(Supplement_5):v222-v222

Publication date:

11th Nov 2025

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