Neuroinflammation and immunothrombosis transcriptional patterns in CCM lesions revealed by spatial transcriptomics

Cerebral cavernous malformation (CCM) immunothrombosis is the connection between immune cells, platelet activation, coagulation cascades and astrocyte-CCM endothelium interaction, and its excessive activation may contribute to neurological disabilities in CCM disease. We characterized the spatial organization of CCM immunothrombosis observed in Pdcd10BECKObrains under normoxic conditions using the murine 10X Genomics Visium Spatial Gene Expression platform. Overall design: Brain endothelial cell-specific conditional Pdcd10-knockout mice were generated by crossing a brain endothelial tamoxifen-regulated Cre recombinase (Slco1c1-iCreERT2) strain with loxP-flanked Pdcd10 (Slco1c1-iCreERT2; Pdcd10fl/fl). On a postnatal day 1 (P1), mice were administered 50 µg of 4-hydroxy-tamoxifen (H7904, Sigma-Aldrich) by intragastric injection to induce genetic inactivation of endothelial CCM3 gene in littermates with Slco1c1-iCreERT2;Pdcd10fl/fl (Pdcd10BECKO) and Pdcd10fl/fl mice were used as littermate controls. P65 Pdcd10BECKO mice, with exacerbated lesions under normoxic conditions was used for analysis.

脑海绵状血管畸形（Cerebral cavernous malformation, CCM）免疫血栓形成是免疫细胞、血小板活化、凝血级联反应与星形胶质细胞-CCM内皮细胞相互作用之间的关联，其过度激活可能导致CCM疾病患者出现神经功能残疾。本研究利用小鼠10X Genomics Visium空间基因表达平台，对常氧条件下Pdcd10BECKO小鼠脑中观察到的CCM免疫血栓形成的空间组织特征进行了表征。实验设计：通过将脑内皮细胞他莫昔芬诱导型Cre重组酶（Slco1c1-iCreERT2）品系与携带loxP侧翼Pdcd10基因的小鼠品系杂交，构建脑内皮细胞特异性条件性Pdcd10敲除小鼠（Slco1c1-iCreERT2; Pdcd10fl/fl，即Pdcd10BECKO）。于出生后第1天（P1），通过胃内注射给予50 μg 4-羟基他莫昔芬（4-hydroxy-tamoxifen，H7904，Sigma-Aldrich），以诱导同窝出生的Slco1c1-iCreERT2;Pdcd10fl/fl（Pdcd10BECKO）小鼠的内皮细胞CCM3基因发生遗传失活；以同窝出生的Pdcd10fl/fl小鼠作为对照。选取常氧条件下病变程度加重的P65龄Pdcd10BECKO小鼠用于后续分析。