CNSC-20. MONOSYNAPTIC TRACING DEFINES BRAIN-WIDE CIRCUIT CONNECTIVITY OF HUMAN GLIOBLASTOMA

Abstract Glioblastoma (GBM), a deadly brain cancer, infiltrates the brain and can be synaptically innervated by neurons. Synaptic inputs onto GBM cells identified so far are largely short-range and glutamatergic in nature. The extent of integration of GBM cells into the brain-wide neuronal circuitry is therefore not well understood. We report the application of transsynaptic viral tracing approaches to study the neuronal connectome of GBM. We applied rabies virus-mediated retrograde tracing and herpes simplex virus (HSV)-mediated anterograde tracing approaches to characterize presynaptic partners in vivo in the adult mouse brain and ex vivo with iPSC-derived neurons, cortical organoid-GBM assembloids, and human surgical specimens. After transplantation into adult mice, GBM cells derived from multiple patients rapidly integrated into brain-wide neuronal circuits. GBM exhibited increased connectivity rates compared to transplanted neural progenitor cells, highlighting functional connectivity as a hallmark of malignant cells. Beyond glutamatergic inputs, we identified neuromodulatory inputs across the brain, including cholinergic inputs from the basal forebrain. We validated these long-range cholinergic neuron-to-glioma synapses signaling through the metabotropic CHRM3 receptor by high-resolution microscopy, anterograde monosynaptic HSV tracing, calcium imaging, and patch-clamp electrophysiology. To interrogate the functional effects of acetylcholine (ACh) on GBM, we performed calcium imaging and RNA sequencing analyses, which showed that ACh stimulation induced sustained calcium oscillations and long-lasting transcriptional reprogramming of GBM cells into a more invasive state via CHRM3. Importantly, CHRM3 activation promoted GBM cell invasion, whereas CHRM3 downregulation suppressed GBM cell invasion in vitro and in vivo, suggesting CHRM3 as a potential therapeutic target for this disease. Together, these results reveal the capacity of human GBM cells to robustly integrate into anatomically and molecularly diverse neuronal circuitry in the adult brain. They also support a model wherein rapid synapse formation onto GBM cells may promote a long-lasting increase in tumor cell fitness.

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Journal
Neuro-Oncology
Published
2024-11-01
DOI
https://doi.org/10.1093/neuonc/noae165.0176
Primary Topic
Neuroscience and Neural Engineering
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article
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article

CNSC-20. MONOSYNAPTIC TRACING DEFINES BRAIN-WIDE CIRCUIT CONNECTIVITY OF HUMAN GLIOBLASTOMA

Guo‐li Ming, Tiago Gonçalves, Jamie Galanaugh, Benjamin C. Kennedy et al.
Neuro-Oncology
Neuroscience and Neural Engineering
article

CNSC-20. MONOSYNAPTIC TRACING DEFINES BRAIN-WIDE CIRCUIT CONNECTIVITY OF HUMAN GLIOBLASTOMA

Guo‐li Ming, Tiago Gonçalves, Jamie Galanaugh, Benjamin C. Kennedy, Daniel H. Geschwind, Katherine L. Helbig, Janardhan P. Bhattarai, Kristen Park, Angela N. Viaene, Weifan Dong, Zev A. Binder, Ahmet Höke, Minghong Ma, Zhuhao Wu, Keerthi Thirtamara Rajamani, MacLean P. Nasrallah, Shang Mu, Yusha Sun, Emily Ling-Lin Pai, Riki Kawaguchi, Sudha Kilaru Kessler, Kimberly M. Christian, Donald M. O’Rourke, Marc V. Fuccillo, Nicolas Toni, Abhijeet Sambangi, Garrett P. Wheeler, Xin Wang, Sang Hoon Kim, Feng Zhang, Qing Wang, Yingqi Wang, Huadong Wang, Yan Hong, Sara Stone, Daniel Zhang, Qian Yang, Hongjun Song, Zhijian Zhang, Isaac Chen, Fuqiang Xu
article en

Abstract

Abstract Glioblastoma (GBM), a deadly brain cancer, infiltrates the brain and can be synaptically innervated by neurons. Synaptic inputs onto GBM cells identified so far are largely short-range and glutamatergic in nature. The extent of integration of GBM cells into the brain-wide neuronal circuitry is therefore not well understood. We report the application of transsynaptic viral tracing approaches to study the neuronal connectome of GBM. We applied rabies virus-mediated retrograde tracing and herpes simplex virus (HSV)-mediated anterograde tracing approaches to characterize presynaptic partners in vivo in the adult mouse brain and ex vivo with iPSC-derived neurons, cortical organoid-GBM assembloids, and human surgical specimens. After transplantation into adult mice, GBM cells derived from multiple patients rapidly integrated into brain-wide neuronal circuits. GBM exhibited increased connectivity rates compared to transplanted neural progenitor cells, highlighting functional connectivity as a hallmark of malignant cells. Beyond glutamatergic inputs, we identified neuromodulatory inputs across the brain, including cholinergic inputs from the basal forebrain. We validated these long-range cholinergic neuron-to-glioma synapses signaling through the metabotropic CHRM3 receptor by high-resolution microscopy, anterograde monosynaptic HSV tracing, calcium imaging, and patch-clamp electrophysiology. To interrogate the functional effects of acetylcholine (ACh) on GBM, we performed calcium imaging and RNA sequencing analyses, which showed that ACh stimulation induced sustained calcium oscillations and long-lasting transcriptional reprogramming of GBM cells into a more invasive state via CHRM3. Importantly, CHRM3 activation promoted GBM cell invasion, whereas CHRM3 downregulation suppressed GBM cell invasion in vitro and in vivo, suggesting CHRM3 as a potential therapeutic target for this disease. Together, these results reveal the capacity of human GBM cells to robustly integrate into anatomically and molecularly diverse neuronal circuitry in the adult brain. They also support a model wherein rapid synapse formation onto GBM cells may promote a long-lasting increase in tumor cell fitness.

Neuro-OncologyVol. 26(Supplement_8)
Albert Einstein College of Medicine (US), Children's Hospital of Philadelphia (US), Johns Hopkins University (US), University of California, Los Angeles (US), Cornell University (US), Thomas Jefferson University Hospital (US), Johns Hopkins Medicine (US), Shenzhen Institutes of Advanced Technology (CN), Weill Cornell Medicine (US), University of Pennsylvania (US), University of Lausanne (CH)
Good health and well-being
Openalex Percentile: Top 79%
Neuroscience and Neural Engineering
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