Lipid Nanoparticle Formulations for Enhanced Co-delivery of siRNA and mRNA
Lipid Nanoparticle Formulations for Enhanced Co-delivery of siRNA and mRNA
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DOI：
10.1021/acs.nanolett.8b01101
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发表时间：
2018-06-01
期刊：
影响因子：
10.8
通讯作者：
Whitehead, Kathryn A.
中科院分区：
文献类型：
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作者：
Ball, Rebecca L.;Hajj, Khalid A.;Whitehead, Kathryn A.
虽然mRNA和siRNA具有显著的治疗潜力，但它们的同时递送先前尚未被探索。为了促进与异常基因上调和下调相关的疾病的治疗，我们寻求在单一类胡萝卜素纳米颗粒（LNP）制剂中共配制siRNA和mRNA。我们将mRNA和siRNA的不同分子特征容纳在由可电离和可生物降解的含胺类胡萝卜素、胆固醇、DSPC、DOPE和PEG-脂质组成的制剂中。令人惊讶的是，siRNA和mRNA在同一LNP中的共制剂增强了两种药物在体外和体内的功效。与仅包封siRNA的LNP相比，共配制的LNP在0.03mg/kg的siRNA剂量下将小鼠中的因子VII基因沉默从44%改善至87%。共配制还改善了mRNA递送，因为与不包括siRNA时相比，与siRNA共配制的0.5mg/kg剂量的mRNA诱导了三倍的荧光素酶蛋白表达。由于并非所有的基因治疗应用都需要两种RNA药物，我们试图将共配制的LNP的益处扩展到仅包封单一类型RNA的制剂。我们通过用带负电荷的聚合物聚苯乙烯磺酸盐（PSS）取代"辅助" RNA来实现这一点。含有PSS的LNP使用少2 - 3倍的RNA介导与标准LNP相同水平的蛋白质沉默或表达。例如，用和不用PSS配制的LNP分别在0.01和0.03mg/kg的siRNA剂量下诱导50%因子VII沉默。总之，这些研究证明了siRNA和mRNA的有效共递送，并显示包含带负电荷的"辅助聚合物"增强了LNP递送系统的功效。
Although mRNA and siRNA have significant therapeutic potential, their simultaneous delivery has not been previously explored. To facilitate the treatment of diseases associated with aberrant gene upregulation and downregulation, we sought to co-formulate siRNA and mRNA in a single lipidoid nanoparticle (LNP) formulation. We accommodated the distinct molecular characteristics of mRNA and siRNA in a formulation consisting of an ionizable and biodegradable amine-containing lipidoid, cholesterol, DSPC, DOPE, and PEG-lipid. Surprisingly, the co-formulation of siRNA and mRNA in the same LNP enhanced the efficacy of both drugs in vitro and in vivo. Compared to LNPs encapsulating siRNA only, co-formulated LNPs improved Factor VII gene silencing in mice from 44 to 87% at an siRNA dose of 0.03 mg/kg. Co-formulation also improved mRNA delivery, as a 0.5 mg/kg dose of mRNA co-formulated with siRNA induced three times the luciferase protein expression compared to when siRNA was not included. As not all gene therapy applications require both RNA drugs, we sought to extend the benefit of co-formulated LNPs to formulations encapsulating only a single type of RNA. We accomplished this by substituting the "helper" RNA with a negatively charged polymer, polystyrenesulfonate (PSS). LNPs containing PSS mediated the same level of protein silencing or expression as standard LNPs using 2-3-fold less RNA. For example, LNPs formulated with and without PSS induced 50% Factor VII silencing at siRNA doses of 0.01 and 0.03 mg/kg, respectively. Together, these studies demonstrate potent co-delivery of siRNA and mRNA and show that inclusion of a negatively charged "helper polymer" enhances the efficacy of LNP delivery systems.