Cancer Neuroscience

Glioblastoma (GB) forms functional synaptic connections with neurons in the tumor microenvironment (TME), contributing to tumor progression, invasion, and resistance to therapy through the formation of tumor microtubes (TMs) (1-2). Dissecting these human neuron-glioma interactions requires high-resolution spatiotemporal analysis to understand their dynamic evolution. We introduce a novel, multiomics workflow that integrates functional and molecular insights to provide a deeper understanding of these dynamics in vitro. As previously described (3), we co-culture human glioblastoma cells with NGN2-iPSC-derived human neurons and track neuron-tumor connectivity over several days using fluorescence and morphological readouts. We will then aim to functionally investigate the effect of tumor cell invasion on the intrinsic electrical activity of neuronal networks and link these measurements with multiomics readouts. 

This functional assessment will be validated and complemented by our COMET™-based multiomics workflow which enables simultaneous, spatially resolved detection of RNA and protein on the same co-culture sections across defined time points. The approach integrates high-plex RNAscope (4) with sequential immunofluorescence (seqIF™, 5) to generate a multimodal molecular map of neuron–glioma interactions. The RNA panel includes markers of mature excitatory neurons (RBFOX3, SLC17A7/VGLUT1), synaptic components (SYP, DLG4), and TM-associated genes (TUBB3, TTYH1). To investigate tumor-induced alterations in neuronal function, we additionally target ion channel-related transcripts (GRIN2B, CACNA1C) and immediate early genes (FOS, ARC), which reflect activity-dependent signaling. The seqIF™ assay complements the transcriptomic layer by detecting proteins associated with TMs (GAP43, Connexin 43, Nestin), synaptic structure (Synaptophysin, PSD95), and glioma identity (OLIG2, SOX2). This integrated workflow enables high-resolution profiling of molecular heterogeneity in both tumor and neuronal compartments, offering novel insights into how glioma invasion perturbs neural network architecture and activity. Ultimately, we aim to establish a reproducible and versatile platform to advance cancer neuroscience and inform therapies that target the nervous system’s pro-tumorigenic influence. 

Spatial biology is transforming our understanding of how the nervous system interacts with cancer.
This talk features Lunaphore’s end-to-end solutions for spatial biology and highlights the application of these tools in neuron-tumor interactions. 

 Daniel Azorín, from the Moor’s lab at the ETH, will present a spatial multiomics workflow that is fully automated on the COMET™ platform, to enable the simultaneous profiling of RNA and protein. While proof-of-concept studies focus on glioblastoma (GB), a cancer that forms functional synaptic connections with neurons and leverages tumor microtubes (TMs) to drive progression, the research group has also demonstrated the versatility of this workflow in mouse models of colorectal cancer (CRC) liver metastases, where neural signaling may play a role in metastatic growth and therapeutic resistance.
Using glioblastoma mouse models and patient-derived tumoroids, tumor heterogeneity, neuronal infiltration, and TM-associated networks are mapped at a single-cell resolution. Complementary co-culture models of human glioblastoma cells and NGN2-iPSC-derived neurons allow for the functional tracking of tumor invasion and its effects on neuronal network activity over time. In parallel, applying the same workflow to CRC liver metastasis models highlights its broader potential to uncover tumor–neural interactions across distinct cancer contexts. 

By integrating RNAscope™ with sequential immunofluorescence (seqIF™), this platform generates multiomics molecular maps linking synaptic gene expression, TM formation, and neuronal activity markers. Together, these studies bridge spatial biology and functional neuroscience, offering a versatile and generalizable framework for Cancer Neuroscience. Ultimately, this approach provides insights into how diverse cancers exploit the nervous system and strategies to target the nervous system’s pro-tumorigenic influence.