Effective prevention of atherosclerosis by non-viral delivery of CRISPR/Cas9
Effective prevention of atherosclerosis by non-viral delivery of CRISPR/Cas9
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DOI：
10.1016/j.nantod.2023.102097
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发表时间：
2024-02
期刊：
影响因子：
17.4
通讯作者：
Chen Xu;Xiaoping Zhang;Weijie Yang;Shijuan Gao;N. Zhao;Ping Li;Jie Du;Yulin Li;Fu‐Jian Xu
中科院分区：
文献类型：
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作者：
Chen Xu;Xiaoping Zhang;Weijie Yang;Shijuan Gao;N. Zhao;Ping Li;Jie Du;Yulin Li;Fu‐Jian Xu
动脉粥样硬化（AS）是导致心血管疾病高发病率和高死亡率的重要原因之一。有效预防AS的治疗选择有限。最近，基于成簇、规则间隔、短回文重复/CRISPR相关核酸酶9（CRISPR/Cas9）的基因组编辑技术显示出永久沉默Pcsk 9基因以降低胆固醇水平并预防AS的巨大潜力。然而，由于基因编辑能力差和出现的基因脱靶，将CRISPR/Cas9系统有效地递送到肝脏用于AS的体内基因编辑仍然是一个挑战。在此，我们报道了一种由低毒和肝靶向聚阳离子（基于普鲁兰糖的乙醇胺修饰的聚甲基丙烯酸缩水甘油酯，PuPGEA）组成的非病毒CRISPR/Cas9递送纳米系统（PuPGEA/pCas 9-Pcsk 9），其可以有效地递送质粒pCas 9-Pcsk 9用于敲除Pcsk 9基因。PuPGEA/pCas 9-Pcsk 9纳米系统在体外可有效敲除小鼠胚胎肝细胞（BNL CL.2）中的Pcsk 9。特别地，与对照组（没有多糖的线性聚阳离子）相比，PuPGEA/pCas 9-Pcsk 9纳米系统可以有效靶向肝脏并成功编辑Pcsk 9，这有助于有效降低胆固醇水平。这种CRISPR/Cas9系统的靶向治疗模式在预防AS过程中表现出令人印象深刻的性能，为各种遗传性肝病和血管疾病的CRISPR/Cas9治疗系统的设计提供了程序化策略。
Atherosclerosis (AS) is one of the most critical reasons behind the high morbidity and mortality of cardiovascular diseases. Limited therapeutic options are available to effectively preventing AS. Recently, clustered, regularly interspaced, short palindromic repeat/CRISPR-associated nuclease 9 (CRISPR/Cas9)-based genome-editing technology has shown great potential to permanently silence Pcsk9 gene to decrease cholesterol level and prevent AS. However, due to the poor ability of gene editing and the arising gene off-target, delivering the CRISPR/Cas9 system effectively to the liver for in vivo gene editing of AS is still a challenge. Herein, we reported a non-viral CRISPR/Cas9 delivery nanosystem (PuPGEA/pCas9-Pcsk9) composed of low-toxic and liver-targeting polycation (pullulan-based ethanolamine-modified poly(glycidyl methacrylate), PuPGEA), which can efficiently deliver a plasmid pCas9-Pcsk9 for the knockout ofPcsk9gene. PuPGEA/pCas9-Pcsk9nanosystem can effectively knock outPcsk9in embryonic liver cells of mice (BNL CL.2) in vitro. Particularly, compared with the control group (linear polycationic without polysaccharide), PuPGEA/pCas9-Pcsk9nanosystems can effectively target liver and successfully editPcsk9, which contributed to the effective decrease of cholesterol level. Such a targeted therapeutic modality with CRISPR/Cas9 system exhibits impressive performances in preventing the process of AS, providing a programmed strategy for the design of CRISPR/Cas9 treatment system for various genetic liver diseases and vascular diseases.