Characterizing and targeting glioblastoma neuron-tumor networks with retrograde tracing

Glioblastomas are invasive brain tumors with high therapeutic resistance. Neuron-to-glioma synapses have been shown to promote glioblastoma progression. However, a characterization of tumor-connected neurons has been hampered by a lack of technologies. Here, we adapted retrograde tracing using rabies viruses to investigate and manipulate neuron-tumor networks. Glioblastoma rapidly integrated into neural circuits across the brain, engaging in widespread functional communication, with cholinergic neurons driving glioblastoma invasion. We uncovered patient-specific and tumor-cell-state-dependent differences in synaptogenic gene expression associated with neuron-tumor connectivity and subsequent invasiveness. Importantly, radiotherapy enhanced neuron-tumor connectivity by increased neuronal activity. In turn, simultaneous neuronal activity inhibition and radiotherapy showed increased therapeutic effects, indicative of a role for neuron-to-glioma synapses in contributing to therapeutic resistance. Lastly, rabies-mediated genetic ablation of tumor-connected neurons halted glioblastoma progression, offering a viral strategy to tackle glioblastoma. Together, this study provides a framework to comprehensively characterize neuron-tumor networks and target glioblastoma. • Rabies-based retrograde tracing maps brain-wide neuronal connectome of glioblastoma • Rapid neuronal integration of glioblastoma precedes neuronal dysfunction • Radiotherapy and perampanel show additive effects by disrupting neuron-tumor networks • Rabies-mediated genetic ablation of neurons halts glioblastoma progression Rapid integration of glioblastoma into neuronal circuits promotes glioblastoma invasion and growth, as shown by rabies-based retrograde tracing. Targeting of neuron-tumor networks with pharmacological or rabies-mediated disconnection offers therapeutic potential.