Rational design of engineered H-ferritin nanoparticles with improved siRNA delivery efficacy across an in vitro model of the mouse BBB
Rational design of engineered H-ferritin nanoparticles with improved siRNA delivery efficacy across an in vitro model of the mouse BBB
复制标题
合理设计工程化 H-铁蛋白纳米粒子，在小鼠 BBB 体外模型中提高 siRNA 递送效率
DOI：
10.1039/d1nr07880a
复制
发表时间：
2022
期刊：
影响因子：
6.7
通讯作者：
Zhang, Xue-Qing
中科院分区：
文献类型：
--
作者：
Yuan, Ziwei;Wang, Bin;Teng, Yilong;Ho, William;Hu, Bin;Boakye-Yiadom, Kofi Oti;Xu, Xiaoyang;Zhang, Xue-Qing
基因疗法在治疗阿尔茨海默病（AD）、中风、神经胶质瘤和帕金森病等不可治愈的脑部疾病方面具有巨大的潜力。主要挑战是缺乏穿过血脑屏障（BBB）的有效基因传递系统，因为大脑中存在复杂的微血管，限制了循环血液中的物质通过。最近，人们越来越努力开发用于脑相关疾病治疗的有前景的基因载体。其中一项进展是自组装重链铁蛋白 (HFn) 纳米颗粒 (NP)。 HFn NP 具有独特的中空球形结构，可以封装核酸药物 (NAD)，并通过与其表面过表达的转铁蛋白受体 1 (TfR1) 相互作用，特异性结合癌细胞和 BBB 内皮细胞 (BBB EC)，从而增加通过 BBB 的摄取。然而，HFn的基因负载能力受到其有限的内部体积和带负电的内表面的限制；因此，这些弊端促使人们需要重塑HFn结构的策略。在这项工作中，我们使用 Chimera 软件 (v 1.14) 分析了 HFn 的三维 (3D) 结构，并通过 HFn 腔表面的精氨酸突变开发了一类内部阳离子 HFn 变体 (HFn+ NP)。这些 HFn+ NPs 在其空腔中呈现出强大的静电力，并且表现出比原始 HFn 更高的基因封装效率。表现最好的候选物 HFn2 在穿过 BBB 后有效地将 siRNA 递送至神经胶质瘤细胞，并在 HFn+ NP 中实现了最高的沉默功效。总的来说，我们的研究结果表明，通过这种基因工程方法获得的 HFn+ NPs 为具有 BBB 穿越能力的核酸递送载体的未来发展提供了重要的见解。
Gene therapy holds tremendous potential for the treatment of incurable brain diseases including Alzheimer's disease (AD), stroke, glioma, and Parkinson's disease. The main challenge is the lack of effective gene delivery systems traversing the blood–brain barrier (BBB), due to the complex microvessels present in the brain which restrict substances from the circulating blood passing through. Recently, increasing efforts have been made to develop promising gene carriers for brain-related disease therapies. One such development is the self-assembled heavy chain ferritin (HFn) nanoparticles (NPs). HFn NPs have a unique hollow spherical structure that can encapsulate nucleic acid drugs (NADs) and specifically bind to cancer cells and BBB endothelial cells (BBB ECs) via interactions with the transferrin receptor 1 (TfR1) overexpressed on their surfaces, which increases uptake through the BBB. However, the gene-loading capacity of HFn is restricted by its limited interior volume and negatively charged inner surface; therefore, these drawbacks have prompted the demand for strategies to remould the structure of HFn. In this work, we analyzed the three-dimensional (3D) structure of HFn using Chimera software (v 1.14) and developed a class of internally cationic HFn variants (HFn+ NPs) through arginine mutation on the lumenal surface of HFn. These HFn+ NPs presented powerful electrostatic forces in their cavities, and exhibited higher gene encapsulation efficacy than naive HFn. The top-performing candidate, HFn2, effectively delivered siRNA to glioma cells after traversing the BBB and achieved the highest silencing efficacy among HFn+ NPs. Overall, our findings demonstrate that HFn+ NPs obtained by this genetic engineering method provide critical insights into the future development of nucleic acid delivery carriers with BBB-crossing ability.