Gene expression profile of microglia following traumatic brain injury and nasal anti-CD3 treatment at acute and chronic timepoints. Mus musculus

Bulk RNA-sequencing was performed to characterize the gene expression profile of microglia at acute and chronic timepoints following traumatic brain injury and nasal anti-CD3 treatment. We further investigated how the chronic microglial transcriptomic profile is modulated following traumatic brain injury and nasal anti-CD3 treatment in female mice with severe TBI, and in male mice with a delayed administration of treatment post-injury. Overall design: Bulk RNA-sequencing analysis was performed on sorted microglia single cell suspensions from the ipsilateral hemisphere of the mouse brains using the microglia-specific 4D4+ antibody at 7 days and 30 days following traumatic brain injury. Preparation for RNA libraries for sequencing was performed using standard Illumina protocols. For the delayed treatment experiment, the therapeutic window of nasal anti-CD3 treatment was delayed to 3 days following injury.

本研究采用批量RNA测序（Bulk RNA-sequencing）技术，对创伤性脑损伤（traumatic brain injury, TBI）联合鼻腔抗CD3治疗后不同急性、慢性时间点下的小胶质细胞（microglia）基因表达谱进行系统表征。我们进一步针对重度TBI雌性小鼠，以及伤后延迟给药的雄性小鼠，探究了创伤性脑损伤联合鼻腔抗CD3治疗后其慢性期小胶质细胞转录组谱的调控变化。整体实验设计：本研究于创伤性脑损伤后第7天和第30天，利用小胶质细胞特异性4D4+抗体，从小鼠大脑损伤同侧半球的单细胞悬液中分选得到小胶质细胞，随后开展批量RNA测序分析。测序所需的RNA文库构建严格遵循标准Illumina实验流程。针对延迟给药实验，鼻腔抗CD3治疗的给药窗口被延迟至伤后第3天。