Cell-specific roles of ephrin-B2 signaling in neurovascular repair and regeneration after ischemic injury of the brain

肝配蛋白 B2 信号在脑缺血损伤后神经血管修复和再生中的细胞特异性作用

• 批准号: 289426136
• 负责人: Professor Dr. Peter Vajkoczy
• 金额: --
• 依托单位: Klinik für Neurochirurgie mit Arbeitsbereich Pädiatrische Neurochirurgie (CVK)
• 依托单位国家: 德国
• 项目类别: Research Units
• 财政年份: 2016
• 资助国家: 德国
• 起止时间: 2015-12-31 至 2021-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/289426136?language=en
• 关键词: Cell; specific; roles; ephrin; B2

A major complication after stroke is the disruption of the blood-brain barrier and development of malignant brain swelling. The brain is capable of repairing its neurovascular unit after stroke; however, often too slow and insufficient. We aim at a better understanding of the underlying mechanisms/therapeutic optimization of this repair. In the 1st funding period, we have revealed that ephrin-B2 is cell-specifically involved in diverse repair processes and that ephrin-B2 activation enhances regeneration of the neurovascular unit after stroke. In the 2nd funding period, we will further strengthen our knowledge on the role of ephrin-B2 in ischemic stroke. To this end, we will more realistically model the clinical scenario, with aged animals and animals with comorbidity phenotypes. We will test whether the relevance of endothelial ephrin-B2 in mediating vascular integrity and BBB repair after stroke is conserved in these animals. Second, we will elucidate the ephrin-B2 dependent signaling pathways and their influence on neurovascular repair and regeneration after stroke, using our genetic models with cell-specific deletion of Efnb2. We will apply RNA-seq technologies and validate the results on the RNA, protein, and functional level. Third, we will unravel the role of neuronal ephrin-B2 in spine biology under normal and ischemic conditions. For this, we will take ex vivo (organotypic slice cultures) and in vivo (2P-LSCM) approaches to visualize changes in spine morphology/number after neuron-specific deletion of Efnb2. Forth, we will apply our knowledge in intravital imaging of the ischemic brain, using 2P-LSCM and cell specific in vivo labeling techniques, to understand the role of cell-specific ephrin-B2 for cell interactions during the anatomical and functional re-assembly of the neurovascular unit. Taken together, this proposal will significantly contribute to our understanding of the endogenous repair mechanisms of the neurovascular unit and the functional/therapeutic role of ephrin-B2 in this context after stroke.

中风后的一个主要并发症是血脑屏障的破坏和恶性脑肿胀的发展。中风后大脑能够修复其神经血管单位;然而，往往太慢和不足。我们的目标是更好地了解这种修复的潜在机制/治疗优化。在第一个资助期，我们发现ephrin-B2是细胞特异性参与不同的修复过程，ephrin-B2激活增强中风后神经血管单位的再生。在第二个资助期，我们将进一步加强对ephrin-B2在缺血性卒中中作用的认识。为此，我们将更现实地模拟临床情况，包括老年动物和具有共病表型的动物。我们将测试内皮ephrin-B2在介导血管完整性和脑血屏障修复中风后的相关性是否是保守的，在这些动物。其次，我们将阐明肝配蛋白B2依赖的信号通路及其对中风后神经血管修复和再生的影响，使用我们的细胞特异性Efnb 2缺失的遗传模型。我们将应用RNA-seq技术，并在RNA，蛋白质和功能水平上验证结果。第三，我们将阐明神经元ephrin-B2在正常和缺血条件下脊柱生物学中的作用。为此，我们将采取离体（器官型切片培养物）和体内（2 P-LSCM）方法来观察神经元特异性缺失Efnb 2后脊柱形态/数量的变化。第四，我们将应用我们的知识在缺血脑的活体成像，使用2 P-LSCM和细胞特异性在体内标记技术，了解细胞特异性ephrin-B2在神经血管单位的解剖和功能重组过程中的细胞相互作用的作用。两者合计，这一建议将显着有助于我们了解内源性修复机制的神经血管单位和功能/治疗作用的ephrin-B2在这种情况下中风后。