Expression profiling in brain microvascular endothelial cells and astrocytes in mouse models of Cranial Cavernous Malformation disease

Cerebral Cavernous Malformations (CCMs) are neurovascular lesions caused by loss-of-function mutations in one of three genes, including KRIT1 (CCM1), CCM2, and PDCD10 (CCM3). Following onset, disease severity ranges from minimal vascular defects to numerous vascular dilations (lesions) distributed throughout the brain. Although the precise pathogenic mechanisms resulting in such a broad range of disease severity are unknown, inter-cellular communication between heterogeneous brain cell types including microvascular endothelial cells, astrocytes, and neurons are likely important in disease progression. To further understand these cell type relationships in the context of CCM, Riboseq was performed in astroctytes and mRNA expression profiling was performed from whole brain extracts as well as isolated brain microvascular endothelial cells. Pdcd10BECKO (Slco1c1-iCreERT2;Pdcd10fl/fl) and Pdcd10fl/fl control mice were crossed with a ribosome tagged mouse line (Aldh1l1-EGFP/Rpl10a). Following CCM induction with tamoxifen administration, brain tissue was harvested. Ribosome-bound RNA was purified from astrocytes and mRNA was purified from whole brain tissue or from purified brain microvascular endothelial cells. Sequencing was then performed on resultant RNA samples.