Extracellular Vesicle-Mediated Delivery of Circular RNA SCMH1 Promotes Functional Recovery in Rodent and Nonhuman Primate Ischemic Stroke Models
Extracellular Vesicle-Mediated Delivery of Circular RNA SCMH1 Promotes Functional Recovery in Rodent and Nonhuman Primate Ischemic Stroke Models
复制标题
细胞外囊泡介导的环状 RNA SCMH1 促进啮齿类和非人灵长类缺血性中风模型的功能恢复
DOI：
10.1161/circulationaha.120.045765
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发表时间：
2020-08-11
期刊：
影响因子：
37.8
通讯作者：
Yao, Honghong
中科院分区：
文献类型：
--
作者：
Yang, Li;Han, Bing;Yao, Honghong
背景资料：中风是成人残疾的主要原因，可严重影响患者的生活质量，但目前还没有有效的药物来加速康复。已知多种环状RNA（circRNA）分子在缺血性脑损伤中起作用。基于慢病毒的表达系统已广泛用于circRNA的基础研究，但此类递送系统的安全性问题限制了对circRNA潜在治疗作用的探索。方法：应用circRNA微阵列技术从急性缺血性脑卒中患者血浆中筛选出circRNA SCMH 1（circSCMH 1）。产生工程化狂犬病病毒糖蛋白-circSCMH 1-细胞外囊泡以选择性地将circSCMH 1递送至脑。尼氏染色检测梗死面积。在啮齿类动物和非人灵长类动物缺血性中风模型中进行行为任务以评价运动功能。高尔基体染色和免疫组化染色检测神经可塑性和胶质细胞活化。蛋白质组学分析和RNA测序数据结合转录谱被用来确定circSCMH 1的下游目标。结果如下：急性缺血性卒中患者血浆中的CircSCMH 1水平显著降低，为预测卒中结局提供了显著的力量。在光血栓性中风小鼠的血浆和梗死周围皮质中进一步证实了circSCMH 1水平的降低。除了证明RNA药物递送技术的概念验证之外，我们还观察到circSCMH 1治疗可以改善小鼠和猴子中风后的功能恢复，并且我们发现circSCMH 1增强了神经元的可塑性并抑制了神经胶质细胞的激活和外周免疫细胞的浸润。CircSCMH 1与转录因子MeCP 2（甲基-CpG结合蛋白2）机械结合，从而释放对MeCP 2靶基因转录的抑制。结论：狂犬病病毒糖蛋白-circSCMH 1-细胞外囊泡通过促进啮齿动物和非人灵长类动物缺血性中风模型的功能恢复提供保护。我们的研究提出了一种潜在的广泛适用的核苷酸药物递送技术，并展示了如何在治疗上利用circRNA来改善卒中后结局的基本机制。
Background: Stroke is a leading cause of adult disability that can severely compromise the quality of life of patients, yet no effective medication currently exists to accelerate rehabilitation. A variety of circular RNA (circRNA) molecules are known to function in ischemic brain injury. Lentivirus-based expression systems have been widely used in basic studies of circRNAs, but safety issues with such delivery systems have limited exploration of the potential therapeutic roles for circRNAs. Methods: Circular RNA SCMH1 (circSCMH1) was screened from the plasma of patients with acute ischemic stroke by using circRNA microarrays. Engineered rabies virus glycoprotein-circSCMH1-extracellular vesicles were generated to selectively deliver circSCMH1 to the brain. Nissl staining was used to examine infarct size. Behavioral tasks were performed to evaluate motor functions in both rodent and nonhuman primate ischemic stroke models. Golgi staining and immunostaining were used to examine neuroplasticity and glial activation. Proteomic assays and RNA-sequencing data combined with transcriptional profiling were used to identify downstream targets of circSCMH1. Results: CircSCMH1 levels were significantly decreased in the plasma of patients with acute ischemic stroke, offering significant power in predicting stroke outcomes. The decreased levels of circSCMH1 were further confirmed in the plasma and peri-infarct cortex of photothrombotic stroke mice. Beyond demonstrating proof-of-concept for an RNA drug delivery technology, we observed that circSCMH1 treatment improved functional recovery after stroke in both mice and monkeys, and we discovered that circSCMH1 enhanced the neuronal plasticity and inhibited glial activation and peripheral immune cell infiltration. CircSCMH1 binds mechanistically to the transcription factor MeCP2 (methyl-CpG binding protein 2), thereby releasing repression of MeCP2 target gene transcription. Conclusions: Rabies virus glycoprotein-circSCMH1-extracellular vesicles afford protection by promoting functional recovery in the rodent and the nonhuman primate ischemic stroke models. Our study presents a potentially widely applicable nucleotide drug delivery technology and demonstrates the basic mechanism of how circRNAs can be therapeutically exploited to improve poststroke outcomes.