2. Academic Validation
  3. Cholinergic neuron-to-glioblastoma synapses in a human iPSC-derived co-culture model

Cholinergic neuron-to-glioblastoma synapses in a human iPSC-derived co-culture model

  • Stem Cell Reports. 2025 Jul 8;20(7):102534. doi: 10.1016/j.stemcr.2025.102534.
Yusha Sun 1,  Xin Wang 2,  Zhijian Zhang 2,  Kristen H Park 1,  Yicheng Wu 2,  Weifan Dong 2,  Daniel Y Zhang 3,  Yao Fu 2,  Feng Zhang 2,  Zev A Binder 4,  Emily Ling-Lin Pai 5,  MacLean P Nasrallah 6,  Kimberly M Christian 2,  Donald M O'Rourke 4,  Nicolas Toni 7,  Guo-Li Ming 8,  Hongjun Song 9
Affiliations

Affiliations

  • 1 Neuroscience Graduate Group, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA.
  • 2 Department of Neuroscience and Mahoney Institute for Neurosciences, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA.
  • 3 Department of Neurosurgery, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA.
  • 4 Department of Neurosurgery, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA; Glioblastoma Translational Center of Excellence, The Abramson Cancer Center, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA.
  • 5 Department of Pathology and Laboratory Medicine, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA.
  • 6 Glioblastoma Translational Center of Excellence, The Abramson Cancer Center, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA; Department of Pathology and Laboratory Medicine, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA.
  • 7 Center for Psychiatric Neurosciences, Lausanne University Hospital, University of Lausanne, Lausanne, Switzerland.
  • 8 Department of Neuroscience and Mahoney Institute for Neurosciences, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA; Institute for Regenerative Medicine, University of Pennsylvania, Philadelphia, PA, USA; Department of Cell and Developmental Biology, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA; Department of Psychiatry, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA. Electronic address: gming@pennmedicine.upenn.edu.
  • 9 Department of Neuroscience and Mahoney Institute for Neurosciences, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA; Department of Neurosurgery, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA; Glioblastoma Translational Center of Excellence, The Abramson Cancer Center, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA; Department of Cell and Developmental Biology, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA; The Epigenetics Institute, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA. Electronic address: shongjun@pennmedicine.upenn.edu.
PMID: 40541171 DOI: 10.1016/j.stemcr.2025.102534
Abstract

Glioblastoma (GBM) integrates extensively into brain-wide neuronal circuits; however, neuron-tumor interactions have largely been studied with glutamatergic neurons in animal models. The role of neuromodulatory circuits for GBM biology in all-human cell systems remains unclear. Here, we report a co-culture system employing patient-derived GBM organoids and human induced pluripotent stem cell (hiPSC)-derived cholinergic neurons. We provided evidence of structural human cholinergic synaptic inputs onto GBM cells via trans-monosynaptic tracing and electron microscopy and functional synaptic interactions through the metabotropic CHRM3 receptor via calcium imaging. Deep single-cell RNA Sequencing of co-cultures compared to GBM monocultures further revealed shifts in tumor transcriptional profiles toward a more proliferative state, with contributions from both diffusible factors and direct contacts, the latter of which are dependent on Cholesterol biosynthesis. Together, our findings support the role of cholinergic inputs in promoting GBM progression and highlight hiPSC-derived co-culture models as a useful platform for Cancer neuroscience.

Keywords

cancer neuroscience; cholinergic neurons; glioblastoma; glioma; human iPSC; neuron-to-glioma synapse; single-cell; synapse; tumor organoids.

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