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Fibronectin rescues aberrant phenotype of endothelial cells lacking either CCM1, CCM2 or CCM3
Author(s) -
Schwefel Konrad,
Spiegler Stefanie,
Kirchmaier Bettina C.,
Dellweg Patricia K. E.,
Much Christiane D.,
PanéFarré Jan,
Strom Tim M.,
Riedel Katharina,
Felbor Ute,
Rath Matthias
Publication year - 2020
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fj.201902888r
Subject(s) - fibronectin , microbiology and biotechnology , extracellular matrix , phenotype , crispr , chemistry , cytoskeleton , actin cytoskeleton , fibril , cell , stress fiber , biology , genetics , focal adhesion , signal transduction , gene
Abstract Loss‐of‐function variants in CCM1/KRIT1 , CCM2 , and CCM3/PDCD10 are associated with autosomal dominant cerebral cavernous malformations (CCMs). CRISPR/Cas9‐mediated CCM3 inactivation in human endothelial cells (ECs) has been shown to induce profound defects in cell‐cell interaction as well as actin cytoskeleton organization. We here show that CCM3 inactivation impairs fibronectin expression and consequently leads to reduced fibers in the extracellular matrix. Despite the complexity and high molecular weight of fibronectin fibrils, our in vitro model allowed us to reveal that fibronectin supplementation restored aberrant spheroid formation as well as altered EC morphology, and suppressed actin stress fiber formation. Yet, fibronectin replacement neither enhanced the stability of tube‐like structures nor inhibited the survival advantage of CCM3 −/− ECs. Importantly, CRISPR/Cas9‐mediated introduction of biallelic loss‐of‐function variants into either CCM1 or CCM2 demonstrated that the impaired production of a functional fibronectin matrix is a common feature of CCM1‐, CCM2‐, and CCM3‐deficient ECs.