Bringing CCM into a dish: cell culture models for cerebral cavernous malformations

Dariush Skowronek; Robin A Pilz; Konrad Schwefel; Christiane D Much; Ute Felbor; Matthias Rath

doi:10.1515/medgen-2021-2091

Bringing CCM into a dish: cell culture models for cerebral cavernous malformations

Med Genet. 2021 Dec 3;33(3):251-259. doi: 10.1515/medgen-2021-2091. eCollection 2021 Sep.

Authors

Dariush Skowronek^{1

2}, Robin A Pilz^{1

2}, Konrad Schwefel^{1

2}, Christiane D Much^{1

2}, Ute Felbor^{1

2}, Matthias Rath^{3

2}

Affiliations

¹ Department of Human Genetics, University Medicine Greifswald, Greifswald, Germany.
² Interfaculty Institute of Genetics and Functional Genomics, University of Greifswald, Greifswald, Germany.
³ Department of Human Genetics, University Medicine Greifswald, Fleischmannstraße 43, D-17475 Greifswald, Germany.

Abstract

Cerebral cavernous malformations (CCMs) are vascular lesions that can cause severe neurological complications due to intracranial hemorrhage. Although the CCM disease genes, CCM1, CCM2, and CCM3, have been known for more than 15 years now, our understanding of CCM pathogenesis is still incomplete. CCM research currently focuses on three main disease mechanisms: (1) clonal expansion of endothelial cells with biallelic inactivation of CCM1, CCM2, or CCM3, (2) recruitment of cells with preserved CCM protein expression into the growing lesion, and (3) disruption of endothelial cell-cell junctions in CCMs. We here describe novel CRISPR/Cas9-based in vitro models of CCM and discuss their strengths and limitations in the context of high-throughput drug screening and repurposing approaches.

Keywords: CRISPR/Cas9 genome editing; cell junctions; cerebral cavernous malformations; human endothelial cells; spheroid sprouting.

Grants and funding

This work was supported by the German Research Foundation (DFG RA2876/2-2), the German Federal Ministry of Education and Research (BMBF; grant number 161L0276), and the Research Association Molecular Medicine of the University Medicine Greifswald (FVMM, grant numbers FOVB-2021-07, FOVB-2020-01, FOVB-2019-01, and FOVB-2018-06). MR is supported by a scholarship from the Gerhard Domagk program of the University Medicine Greifswald.