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Co-culture of Glioblastoma Stem-like Cells on Patterned Neurons to Study Migration and Cellular Interactions

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Long-range cholinergic input promotes glioblastoma progression.

Yang Yang¹, Chuanyan Yang², Xuezhu Chen³

¹Institute of Pathology & Southwest Cancer Center, Glioma Medical Research Center, Southwest Hospital, Third Military Medical University (Army Medical University), and The Key Laboratory of Tumor Immunopathology, The Ministry of Education of China, Chongqing 400038, China; Department of Neurosurgery, Glioma Medical Research Center, Southwest Hospital, Third Military Medical University (Army Medical University), Chongqing 400038, China; Department of Neurosurgery, No.904 Hospital, Anhui Medical University, Wuxi 214044, China.

|August 19, 2025

Summary

Researchers mapped brain tumor connections, finding that while local inputs are often glutamatergic, long-range connections, including cholinergic pathways, significantly influence glioblastoma (GBM) progression. Blocking these pathways shows therapeutic promise.

Keywords:

CHRM3 cholinergic input diagonal band of Broca donepezil glioblastoma glutamatergic input neural circuit manipulation neuro-glioma interaction rabies virus scopolamine

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Area of Science:

Neuroscience
Oncology
Molecular Biology

Background:

Glioblastoma (GBM) is the most aggressive brain tumor, intricately linked with neural networks.
Understanding the neuron-glioma connectome is crucial, as glutamatergic input influences tumor progression, but broader connectivity remains unclear.

Purpose of the Study:

To map the brain-wide neural input to patient-derived glioblastoma xenografts.
To elucidate the functional role of specific neural inputs in GBM progression.
To identify potential therapeutic targets within neuron-glioma communication pathways.

Main Methods:

Monosynaptic rabies tracing to map neural inputs.
Functional assays assessing acetylcholine release and muscarinic receptor involvement.
Analysis of calcium transients and gene expression in response to glutamatergic and cholinergic signaling.
Pharmacological interventions with anticholinergic and acetylcholinesterase inhibitor drugs.

Main Results:

A consistent organizational logic in neural input to GBM was revealed: local inputs are primarily glutamatergic, while long-range inputs show diverse neurotransmitter profiles.
Basal forebrain cholinergic projections were identified as a conserved input across GBM sites.
Presynaptic acetylcholine release promotes GBM progression via CHRM3 in a circuit-specific manner.
Glutamatergic and cholinergic signals synergistically enhance glioma calcium transients but differentially control gene transcription.
Dual blockade of glutamatergic and cholinergic pathways demonstrated additive anti-tumor effects.

Conclusions:

Neuron-glioma connectivity is complex, involving both local and long-range neuromodulatory pathways.
Cholinergic signaling, particularly through CHRM3, plays a significant role in GBM progression.
Targeting long-range neuromodulatory pathways, such as cholinergic projections, represents a promising therapeutic strategy for glioblastoma.