Cholinergic neuronal activity promotes diffuse midline glioma growth through muscarinic signaling

Richard Drexler; Antonia Drinnenberg; Avishai Gavish; Belgin Yalçin; Kiarash Shamardani; Abigail E Rogers; Rebecca Mancusi; Vrunda Trivedi; Kathryn R Taylor; Yoon Seok Kim; Pamelyn J Woo; Neeraj Soni; Minhui Su; Alexandre Ravel; Eva Tatlock; Alexandra Midler; Samuel H Wu; Charu Ramakrishnan; Ritchie Chen; Alberto E Ayala-Sarmiento; David Rincon Fernandez Pacheco; La'Akea Siverts; Tanya L Daigle; Bosiljka Tasic; Hongkui Zeng; Joshua J Breunig; Karl Deisseroth; Michelle Monje

doi:10.1016/j.cell.2025.05.031

Cholinergic neuronal activity promotes diffuse midline glioma growth through muscarinic signaling

Cell. 2025 Jun 19:S0092-8674(25)00618-X. doi: 10.1016/j.cell.2025.05.031. Online ahead of print.

Authors

Richard Drexler¹, Antonia Drinnenberg², Avishai Gavish³, Belgin Yalçin¹, Kiarash Shamardani¹, Abigail E Rogers¹, Rebecca Mancusi¹, Vrunda Trivedi¹, Kathryn R Taylor¹, Yoon Seok Kim¹, Pamelyn J Woo¹, Neeraj Soni¹, Minhui Su¹, Alexandre Ravel¹, Eva Tatlock¹, Alexandra Midler¹, Samuel H Wu¹, Charu Ramakrishnan⁴, Ritchie Chen⁴, Alberto E Ayala-Sarmiento⁵, David Rincon Fernandez Pacheco⁵, La'Akea Siverts⁶, Tanya L Daigle⁶, Bosiljka Tasic⁶, Hongkui Zeng⁶, Joshua J Breunig⁵, Karl Deisseroth⁷, Michelle Monje⁸

Affiliations

¹ Department of Neurology and Neurological Sciences, Stanford University, Stanford, CA 94305, USA.
² Department of Bioengineering, Stanford University, Stanford, CA 94305, USA.
³ Department of Neurology and Neurological Sciences, Stanford University, Stanford, CA 94305, USA; Howard Hughes Medical Institute, Stanford, CA 94305, USA.
⁴ Department of Bioengineering, Stanford University, Stanford, CA 94305, USA; Department of Psychiatry and Behavioral Sciences, Stanford University, Stanford, CA, USA.
⁵ Board of Governors Regenerative Medicine Institute, Cedars-Sinai Medical Center, Los Angeles, CA 90048, USA.
⁶ Allen Institute for Brain Science, Seattle, WA, USA.
⁷ Department of Bioengineering, Stanford University, Stanford, CA 94305, USA; Department of Psychiatry and Behavioral Sciences, Stanford University, Stanford, CA, USA; Howard Hughes Medical Institute, Stanford, CA 94305, USA. Electronic address: deissero@stanford.edu.
⁸ Department of Neurology and Neurological Sciences, Stanford University, Stanford, CA 94305, USA; Department of Psychiatry and Behavioral Sciences, Stanford University, Stanford, CA, USA; Howard Hughes Medical Institute, Stanford, CA 94305, USA. Electronic address: mmonje@stanford.edu.

PMID: 40541184
DOI: 10.1016/j.cell.2025.05.031

Abstract

Glutamatergic neuronal activity promotes proliferation of both oligodendrocyte precursor cells (OPCs) and gliomas, including diffuse midline glioma (DMG). However, the role of neuromodulatory brainstem neurons projecting to midline structures where DMGs arise remains unexplored. Here, we demonstrate that midbrain cholinergic neuronal activity modulates OPC and DMG proliferation in a circuit-dependent manner. Optogenetic stimulation of the cholinergic pedunculopontine nucleus (PPN) promotes glioma growth in pons, while stimulation of the laterodorsal tegmentum nucleus (LDT) drives proliferation in thalamus. DMG-bearing mice exhibit higher acetylcholine release and increased cholinergic neuronal activity over the disease course. In co-culture, cholinergic neurons enhance DMG proliferation, and acetylcholine directly acts on DMG cells. Single-cell RNA sequencing revealed high CHRM1 and CHRM3 expression in primary DMG samples. Pharmacological or genetic blockade of M1/M3 receptors abolished cholinergic activity-driven DMG proliferation. Taken together, these findings demonstrate that midbrain cholinergic long-range projections promote activity-dependent DMG growth, mirroring a parallel proliferative effect on healthy OPCs.

Keywords: DMG; LDT; OPC; PPN; cholinergic neuron; diffuse midline glioma; glioma; laterodorsal tegmentum nucleus; neuron; neuron-glioma; neuronal activity; oligodendrocyte; oligodendrogenesis; pedunculopontine nucleus.