Artikel
Spatiotemporal diversity of cell communication in glioblastoma aid transcriptional plasticity
Zeitliche und räumlicher Vielfältigkeit zelluläre Kommunikation erhöht die transkriptionelle Plastizität in Glioblastomen
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Autoren
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Kevin Joseph
- Medical Center, University of Freiburg, Translational neuroOncology Research Group, Freiburg, Deutschland; Medical Center, University of Freiburg, Department of Neurosurgery, Freiburg, Deutschland; Medical Center, University of Freiburg, Freiburg, Deutschland
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Lea Vollmer
- Medical Center, University of Freiburg, Translational NeuroOncology Research Group, Freiburg, Deutschland; Medical Center, University of Freiburg, Department of Neurosurgery, Freiburg, Deutschland; Medical Center, University of Freiburg, Freiburg, Deutschland
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Vidhya Madapusi Ravi
- Medical Center, University of Freiburg, Translational NeuroOncology Research Group, Freiburg, Deutschland; Medical Center, University of Freiburg, Department of Neurosurgery, Freiburg, Deutschland; Medical Center, University of Freiburg, Neuroelectronic Systems, Freiburg, Deutschland; Medical Center, University of Freiburg, Freiburg, Deutschland
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Jürgen Beck
- Medical Center, University of Freiburg, Translational NeuroOncology Research Group, Freiburg, Deutschland; Medical Center, University of Freiburg, Department of Neurosurgery, Freiburg, Deutschland; Medical Center, University of Freiburg, Freiburg, Deutschland
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Ulrich Hofmann
- Medical Center, University of Freiburg, Department of Neurosurgery, Freiburg, Deutschland; Medical Center, University of Freiburg, Neuroelectronic Systems, Freiburg, Deutschland; Medical Center, University of Freiburg, Freiburg, Deutschland
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Oliver Schnell
- Medical Center, University of Freiburg, Translational NeuroOncology Research Group, Freiburg, Deutschland; Medical Center, University of Freiburg, Department of Neurosurgery, Freiburg, Deutschland; Medical Center, University of Freiburg, Freiburg, Deutschland
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Dieter Henrik Heiland
- Medical Center, University of Freiburg, Translational NeuroOncology Research Group, Freiburg, Deutschland; Medical Center, University of Freiburg, Department of Neurosurgery, Freiburg, Deutschland; Medical Center, University of Freiburg, Freiburg, Deutschland
| Veröffentlicht: | 4. Juni 2021 |
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Gliederung
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Objective: Owing to recent advances in understanding of the active functional states exhibited within glioblastoma (GBM), intra-tumoral cellular signaling has moved into focus of neuro-oncological research. In our study, we aim to explore the spatial and temporal diversity of transcellular signaling and investigate the correlation to transcriptional dynamics, cell behavior and target molecules.
Methods: Electrophysiological mapping was performed by planar array-based microelectrodes, in addition to calcium imaging in both cell culture and human neocortical section based GBM models. Selective inhibition/activation of selected ion-channels was performed to identify the channels involved in the alteration of cytosolic ionic flux. Paired patch-clamp recordings within the GBM network were used to validate intercellular electrical signaling. Transcriptional dynamics and plasticity were examined by means of scRNA-sequencing with CRISPR based perturbation, spatial transcriptomics and deep long-read RNA-sequencing.
Results: Investigation of electrophysiological profiles of six different cell lines revealed highly variable baseline activity, significantly enriched in cell lines with astrocytic/mesenchymal transcriptional signature. Despite the different characteristics of transcellular communication, all cell-lines followed the rules of scale-free networks, which was confirmed in human neocortical section based GBM model. In the GBM model, a significantly increased level of baseline activity was shown for all cell lines. Cellular signaling was directly correlated to changes in the environment, like hypoxia or acidification. Using single-cell sequencing and proteomics, we identified Neurobeachin (NBEA) which plays a crucial role in synaptogenesis. CRISPR loss-of NBEA resulted in alterations in cellular morphology and decreased cellular connectivity. Electrical signaling was lost in cell culture and significantly reduced in the GBM model. Single-cell sequencing of perturbed tumor cells in the GBM model revealed a loss of neuronal lineages and significant reduction of cellular stress levels (mesenchymal signature).
Conclusion: Our findings highlight the role of electrical signaling in glioblastoma. Cellular stressors induce electrical signaling, leading to transcriptional adaptation suggesting that there exists a highly complex and powerful mechanism for fast dynamic cellular adaptation. The identification of key player, e.g. NBEA, will help to therapeutically target transcellular signaling.
