CNSC-21. CHARACTERIZATION OF NEURON-TUMOR INTERACTIONS USING HUMAN CO-CULTURES

Abstract

Abstract Glioblastoma are incurable brain tumors characterized by their colonization of the entire brain and their notorious therapeutic resistance. Recently, we discovered long membrane tubes called tumor microtubes contributing to invasion, network formation of tumor-tumor networks and therapeutic resistance. Subsequently, heterogeneous networks of neurons and glioblastoma cells were characterized, which can communicate by synaptic and perisynaptic contacts as well as by paracrine mechanisms. Currently used models of studying neuron-glioblastoma interactions are limited by the possibility to study glioblastoma in a defined human neuronal microenvironment. Here, we set out to derive excitatory and inhibitory neurons from embryonic stem cells via lentiviral reprogramming and co-cultured them with patient-derived glioblastoma cells. We could show that structural and functional neuron-glioblastoma synaptic contacts are formed. Functional communication between neurons and glioblastoma cells were characterized with calcium imaging, showing similar complex calcium dynamics previously characterized with in vivo imaging of patient-derived xenograft models. The single-cell glioblastoma morphology was morphometrically similar to that of human glioblastoma tissue. Tumor microtubes and the formation of tumor-tumor networks could be demonstrated. Additionally, glioblastoma invasion patterns in our human neuronal co-culture model resemble invasion patterns recently characterized with patient-derived xenograft models. Lastly, we investigated reciprocal neuron-glioblastoma interactions and longitudinally characterized neuronal activity with patch-clamp electrophysiology. In conclusion, we provide a novel human neuron-glioblastoma co-culture system allowing in-depth molecular and functional characterization for future Cancer Neuroscience studies.