CCM3 knockout and wild-type cells in dialogue: Analyzing and modulating tumor-like mechanisms of cerebral cavernous malformations in vascular mosaic organoid models

• Applicant: Dr. Robin Alexander Pilz
• Subject Area: Human Genetics
• Term: since 2025
• Project identifier: Deutsche Forschungsgemeinschaft (DFG) - Project number 564694454

Cerebral cavernous malformations (CCMs) are hereditary vascular lesions in the brain or spinal cord that grow in number and size throughout life and may cause severe neurological complications due to hemorrhage. Recent studies have shown that, in addition to pronounced endothelial dysfunction, "tumor-like" mechanisms are also involved in the development of CCMs. Intriguingly, CCM3-deficient cells have been observed to proliferate abnormally upon direct contact with wild-type cells. This behavior is particularly interesting because CCM lesions consist of CCM-deficient cells and cells with preserved CCM expression. However, how these cells communicate and interact with each other on a molecular and functional level is poorly understood and will therefore be addressed in more detail in this project. By establishing and using three-dimensional vascular mosaic organoid models, an innovative methodology will be applied to mimic the in vivo situation as closely as possible. The combination of high-content imaging analyses and single-cell RNA sequencing is anticipated to enable new insight into the tumor-like proliferation of CCM3-deficient cells and the cell-cell interactions between knockout and wild-type populations. To better understand the role of signaling pathways involved, CCM3-deficient endothelial cells within the mosaic organoid system will be genetically modified using CRISPR interference technology. Following preliminary evidence, it will be investigated whether disruption of the TGFBR1-mediated signaling pathway can rescue the dysregulated cellular processes. Overall, this project has great potential to improve our understanding of CCM pathogenesis and to identify promising targets for the development of new therapeutic strategies.

• DFG Programme: Research Grants