Synthesis of siRNA nanoparticles to silence plaque-destabilizing gene in atherosclerotic lesional macrophages.
Synthesis of siRNA nanoparticles to silence plaque-destabilizing gene in atherosclerotic lesional macrophages.
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DOI：
10.1038/s41596-021-00665-4
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发表时间：
2022-03
期刊：
影响因子：
14.8
通讯作者：
Tao, Wei
中科院分区：
文献类型：
--
作者：
Huang, Xiangang;Liu, Chuang;Kong, Na;Xiao, Yufen;Yurdagul, Arif, Jr.;Tabas, Ira;Tao, Wei
动脉粥样硬化病变中的巨噬细胞促进斑块进展，是心血管研究中有吸引力的治疗靶点。在这里，我们提出了一个协议的小干扰RNA（siRNA）纳米粒子（NP）的合成，靶向病变巨噬细胞作为一个潜在的治疗动脉粥样硬化。晚期人类和小鼠动脉粥样硬化斑块巨噬细胞中的Ca 2 +/钙调蛋白依赖性蛋白激酶γ（CaMK II γ）活性通过下调巨噬细胞受体MerTK的表达驱动坏死。因此，选择性抑制病变巨噬细胞中的CaMKIIγ对于治疗晚期动脉粥样硬化具有很大的希望。我们最近开发了一种siRNA NP平台，可以选择性地沉默巨噬细胞中的CaMKIIγ，从而增加斑块稳定性。我们提供了一个详细的协议的NP组件的合成，制备和表征（物理化学和体外）的siRNA NPs，并在动脉粥样硬化小鼠体内的siRNA NPs的治疗效果和它们的生物相容性的评价。我们的siRNA负载的聚合物-脂质杂合纳米颗粒通过稳健的自组装方法构建，表现出优异的系统性siRNA递送的体内特征。按照该方案，需要3-5天来制备siRNA NPs，8-10天来表征NPs，4-5周来评估它们在建立的动脉粥样硬化小鼠中的治疗效果。通过改变加载在纳米颗粒中的RNA分子，可以靶向病变巨噬细胞，用于探索和验证动脉粥样硬化中的新靶点/途径。
Macrophages in atherosclerotic lesions promote plaque progression and are an attractive therapeutic target in cardiovascular research. Here we present a protocol for synthesis of small interfering RNA (siRNA) nanoparticles (NP) that target lesional macrophages as a potential treatment for atherosclerosis. Ca2+/calmodulin-dependent protein kinase γ (CaMKIIγ) activity in macrophages of advanced human and mouse atherosclerotic plaques drives necrosis via down-regulating the expression of the efferocytosis receptor MerTK. Therefore, selective inhibition of CaMKIIγ in lesional macrophages holds great promise for the treatment of advanced atherosclerosis. We have recently developed a siRNA NP platform that can selectively silence CaMKIIγ in macrophages, resulting in increased plaque stability. We provide a detailed protocol for the synthesis of NP components, the preparation and characterization (physicochemical and in vitro) of siRNA NPs, and the evaluation of in vivo therapeutic effects of siRNA NPs and their biocompatibility in atherosclerotic mice. Our siRNA-loaded polymer-lipid hybrid NPs are constructed via a robust self-assembly method, exhibiting excellent in vivo features for systemic siRNA delivery. Following this protocol, it takes 3-5 d to prepare the siRNA NPs, 8-10 d to characterize the NPs, and 4-5 weeks to evaluate their therapeutic effects in established atherosclerotic mice. By changing the RNA molecules loaded in the NPs, lesional macrophages can be targeted for the exploration and validation of new targets/pathways in atherosclerosis.
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影响因子：
10.8
作者：
Huang, Xiangang;Wu, Gongwei;Tao, Wei
通讯作者：
Tao, Wei
影响因子：
8.3
作者：
Díez S;Miguéliz I;Tros de Ilarduya C
通讯作者：
Tros de Ilarduya C
影响因子：
46.9
作者：
Gilleron, Jerome;Querbes, William;Zerial, Marino
通讯作者：
Zerial, Marino
影响因子：
64.8
作者：
Castanotto, Daniela;Rossi, John J.
通讯作者：
Rossi, John J.
DOI：
10.1056/nejmoa2035389
发表时间：
2021-02-04
期刊：
The New England journal of medicine
影响因子：
--
作者：
Baden LR;El Sahly HM;Essink B;Kotloff K;Frey S;Novak R;Diemert D;Spector SA;Rouphael N;Creech CB;McGettigan J;Khetan S;Segall N;Solis J;Brosz A;Fierro C;Schwartz H;Neuzil K;Corey L;Gilbert P;Janes H;Follmann D;Marovich M;Mascola J;Polakowski L;Ledgerwood J;Graham BS;Bennett H;Pajon R;Knightly C;Leav B;Deng W;Zhou H;Han S;Ivarsson M;Miller J;Zaks T;COVE Study Group
通讯作者：
COVE Study Group