Thrombospondin1 replacement prevents cerebral cavernos malformations

We have purified primary brain microvascular endothelial cells (BMEC) from mice bearing floxed alleles of Krit1 (Krit1fl/fl) and an endothelial specific tamoxifen-regulated Cre recombinase (Pdgfb-iCreERT2), and used these to delete Krit1 in a time-controlled manner (Krit1ECKO). To elucidate the pathogenesis of cerebral cavernous malformations (CCM) we used genome-wide RNA sequencing (RNA-seq) to characterize the transcriptome of primary BMEC following acute genetic inactivation of Krit1. Treatment of Krit1f/f-Pdgfb-iCreERT2 BMEC with 5 µM-hydroxy-tamoxifen deleted Krit1 (Krit1ECKO) reduced KRIT1 mRNA and protein by >90% within 5 days compared to hydroxy-tamoxifen-treated Krit1fl/fl controls. Three independent biological replicates were used for the analysis.