CCM2L (Cerebral Cavernous Malformation 2 Like) Deletion Aggravates Cerebral Cavernous Malformation Through Map3k3-KLF Signaling Pathway

Background and Purpose: Cerebral cavernous malformation (CCM) is a common cerebrovascular disease. CCMs are major causes of stroke, cerebral hemorrhage, and neurological deficits in young individuals. Loss-of-function mutations inCCM1 ,CCM2 , andCCM3 have been identified to cause CCM in humans.Ccm2-like (Ccm2l ) is a paralog ofCcm2 and is predominantly expressed in endothelial cells (ECs). CCM2L (CCM2-like) competes with CCM2 for binding to CCM1 and has been shown to have an antagonistic function to that of CCM2 during vascular development. The role of CCM2L in CCM pathogenesis is unknown.Methods: We isolated brain ECs from the inducible-CCM mouse models for gene expression analysis. Micro-computed tomography imaging was used to analyze CCM lesion burden from the genetic cross ofCcm2l knockout mice (Ccm2l −/− ) withCcm1 orCcm2 -deficient mice to determine the role ofCcm2l in CCM pathogenesis. Genetic crosses withMap3k3 fl/fl mice were used to determine the role ofMap3k3 inCcm2l -facilitated CCM formation.Results: We demonstrated increasedCcm2l expression in brain ECs ofCcm2 -deficient mice. Analysis of RNA-seq data from CCM patient samples revealed a trend of increasedCCM2L expression and its positive correlation with Kruppel-like factor 2/4(KLF2/4 ) expression. Micro-computed tomography revealed that the deletion ofCcm2l inCcm2 -deficient mice increased CCM lesion volume compared with that of controls but had no effect on lesion numbers. Correlating to the increased lesion burden,Klf2/4 mRNA expressions in brain ECs were significantly increased in double knockouts (Ccm2 - andCcm2l -deficient mice) compared with that of controls (Ccm2 deficient). Hemizygous deletion ofMap3k3 in ECs relieved CCM lesion burden in the double knockouts. These results suggest that CCM2L regulates the Map3k3-KLF signaling pathway in CCM pathogenesis.Conclusions: Loss of CCM2L aggravates CCM lesion formation in theCcm2 -deficient mouse model through increased Map3k3-KLF signaling. Our data suggest that increasedCcm2l expression is a compensatory mechanism in CCM pathogenesis.