PDF(Pubmed)
Abstract：
Necroptosis is implicated in the pathogenesis of ischemic stroke. However, the mechanism underlying the sequential recruitment of receptor-interacting protein kinase 1 (RIPK1) and N-ethylmaleimide-sensitive fusion ATPase (NSF) in initiating necroptosis remains poorly understood, and the role of NSF in ischemic stroke is a subject of controversy. Here, we utilized a recently emerging RNA-targeting CRISPR system known as CasRx, delivered by AAVs, to knockdown Ripk1 mRNA and Nsf mRNA around the ischemic brain tissue. This approach resulted in a reduction in infarct and edema volume, as well as an improvement in neurological deficits assessed by Bederson score, RotaRod test, and Adhesive removal test, which were achieved by RIPK1/receptor-interacting protein kinase 3/mixed lineage kinase domain-like protein signaling pathway involved in neuronal necroptosis. In conclusion, the downregulation of Ripk1 mRNA and Nsf mRNA mediated by CRISPR-CasRx holds promise for future therapeutic applications aimed at ameliorating cerebral lesions and neurological deficits following the ischemic stroke.
摘要：
坏死与缺血性中风的发病机理有关。然而,受体相互作用蛋白激酶1（RIPK1）和N-乙基马来酰亚胺敏感的融合ATP酶（NSF）在启动坏死性凋亡中的顺序募集的机制仍然知之甚少，NSF在缺血性卒中中的作用是一个有争议的话题。这里,我们使用了一种最近出现的RNA靶向CRISPR系统，称为CasRx，由AAV交付，敲除缺血脑组织周围的Ripk1mRNA和NsfmRNA。这种方法导致梗死和水肿体积减少，以及通过Bederson评分评估的神经功能缺损的改善，旋转杆试验,和粘合剂去除试验，RIPK1/受体相互作用蛋白激酶3/混合谱系激酶结构域样蛋白信号通路参与神经元坏死性凋亡。总之,由CRISPR-CasRx介导的Ripk1mRNA和NsfmRNA的下调为未来旨在改善缺血性卒中后脑损伤和神经功能缺损的治疗应用带来了希望.
