Pharmacologic or genetic inhibition of EphB3 receptor kinase impacts CNS pathology during progressive EAE

We evaluated the role of EphB3 on the regulation of astrocyte and microglial responses and its potential as a therapeutic target during progressive EAE. We induced EAE in NOD mice that will develop a progressive form of the disease. Starting at the beginning of the progressive phase, A38 was administered and alternatively EphB3 was KD in astrocytes. Both treatments ameliorated the EAE. The transcriptional analysis of astrocytes and microglia revealed the decreased expression of genes associated to inflammation and neurodegeneration in both conditions. Overall design: Astrocytes and microglia were isolated from CNS of mice undergoing progressive model of EAE and which have been intrathecally injected with lentviral vectors either ctrl, or shEphB3 astrocyte specific. shCtrl were additionally treated with A38 or the vehicle as control.

本研究评估了EphB3在进行性实验性自身免疫性脑脊髓炎（Experimental Autoimmune Encephalomyelitis, EAE）进程中对星形胶质细胞（astrocyte）与小胶质细胞（microglia）应答的调控作用，以及其作为治疗靶点的潜在价值。我们在易出现进行性疾病表型的非肥胖糖尿病（Non-Obese Diabetic, NOD）小鼠中构建EAE模型。自疾病进行性阶段起始之时，分别给予A38给药处理，或在星形胶质细胞中敲低（knockdown, KD）EphB3的表达。两种处理方式均有效缓解了EAE的病情进展。对星形胶质细胞与小胶质细胞的转录组分析结果显示，在两种处理条件下，与炎症反应及神经退行性变相关的基因表达均显著下调。总体实验设计：从罹患进行性EAE模型的小鼠中枢神经系统（Central Nervous System, CNS）中分离星形胶质细胞与小胶质细胞；这些小鼠经鞘内注射了星形胶质细胞特异性慢病毒载体（lentiviral vectors），分别为对照载体（ctrl）或靶向EphB3的短发夹载体（shEphB3）。其中，注射对照短发夹载体（shCtrl）的小鼠还额外接受了A38处理或溶剂对照处理。