Bio-clickable, small extracellular vesicles-COCKTAIL therapy for ischemic stroke
Bio-clickable, small extracellular vesicles-COCKTAIL therapy for ischemic stroke
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DOI：
10.1016/j.jconrel.2023.10.003
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发表时间：
2023-10-11
影响因子：
10.8
通讯作者：
Zhang, Zhijun
中科院分区：
文献类型：
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作者：
Haroon, Khan;Ruan, Huitong;Zhang, Zhijun
通过血脑屏障输送大的治疗分子来治疗缺血性中风仍然具有挑战性。NR2B9c是一种有效的神经保护肽，但它的安全性和靶向性需要一种高效、自然和非免疫原性的递送技术。小细胞外囊泡（sev）作为一种非免疫原性的天然货物递送系统显示出巨大的潜力；然而，需要对其低效的大脑靶向进行调整。本研究通过生物正交点击化学反应将狂犬病毒糖蛋白29与sev表面偶联，然后加载NR2B9c，最终生成脑卒中特异性治疗鸡尾酒（sEVsCOCKTAIL）。体外培养原代神经元和神经2a细胞，体内研究采用短暂性大脑中动脉闭塞模型，评价sEVs-COCKTAIL的神经元靶向性和抗缺血性脑卒中潜能。生物可点击的sev被神经元选择性摄取，而不是神经胶质细胞。在体外缺氧-葡萄糖剥夺缺血性脑卒中模型中，sEVs-COCKTAIL对活性氧和细胞凋亡表现出显著的抑制作用。体内研究进一步证明了生物可点击sev的脑靶向性和半衰期延长，可将NR2B9c输送到缺血脑，减轻脑卒中损伤。sews - cocktail治疗可显著提高短暂性大脑中动脉闭塞后的行为恢复，减少神经元凋亡。NR2B9c被递送到结合突触后密度蛋白-95的神经元，抑制n-甲基- d -天冬氨酸受体介导的氧化应激过度产生，减轻蛋白b细胞淋巴瘤2和P38蛋白的表达。我们的研究结果为靶向递送系统提供了一种高效且具有生物相容性的方法，这是一种很有前途的中风治疗方式。
Delivering large therapeutic molecules via the blood-brain barrier to treat ischemic stroke remains challenging. NR2B9c is a potent neuroprotective peptide but it's safe and targeted delivery to the brain requires an efficient, natural, and non-immunogenic delivery technique. Small extracellular vesicles (sEVs) have shown great potential as a non-immunogenic, natural cargo delivery system; however, tailoring of its inefficient brain targeting is desired. Here, we coupled rabies virus glycoprotein 29 with sEVs surface via bio-orthogonal click chemistry reactions, followed by loading of NR2B9c, ultimately generating stroke-specific therapeutic COCKTAIL (sEVsCOCKTAIL). Primary neurons and Neuro-2a cells were cultured for in vitro and transient middle cerebral artery occlusion model was used for in vivo studies to evaluate neuron targeting and anti-ischemic stroke potential of the sEVs-COCKTAIL. Bio-clickable sEVs were selectively taken up by neurons but not glial cells. In the in vitro ischemic stroke model of oxygen-glucose deprivation, the sEVs-COCKTAIL exhibited remarkable potential against reactive oxygen species and cellular apoptosis. In vivo studies further demonstrated the brain targeting and increased half-life of bio-clickable sEVs, delivering NR2B9c to the ischemic brain and reducing stroke injury. Treatment with the sEVs-COCKTAIL significantly increased behavioral recovery and reduced neuronal apoptosis after transient middle cerebral artery occlusion. NR2B9c was delivered to neurons binding to post-synaptic density protein-95, inhibiting N-methyl-D-Aspartate receptor-mediated over production of oxidative stress and mitigating protein B-cell lymphoma 2 and P38 proteins expression. Our results provide an efficient and biocompatible approach to a targeted delivery system, which is a promising modality for stroke therapy.