Electrostatic adsorption of polyanions onto lipid nanoparticles controls uptake, trafficking, and transfection of RNA and DNA therapies.
Electrostatic adsorption of polyanions onto lipid nanoparticles controls uptake, trafficking, and transfection of RNA and DNA therapies.
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聚阴离子在脂质纳米粒子上的静电吸附控制 RNA 和 DNA 疗法的摄取、运输和转染。
DOI：
10.1073/pnas.2307809121
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发表时间：
2024
影响因子：
11.1
通讯作者：
Hammond,PaulaT
中科院分区：
文献类型：
--
作者：
Nabar,Namita;Dacoba,TamaraG;Covarrubias,Gil;Romero-Cruz,Denisse;Hammond,PaulaT
核酸疗法的快速发展突出了基因疗法的巨大治疗潜力。脂质纳米颗粒（LNPs）是一种高效的非病毒转染剂，可以封装和递送各种核酸治疗药物，包括但不限于信使RNA （mRNA）、沉默RNA （siRNA）和质粒DNA （pDNA）。然而，靶向LNP介导的全身递送的一个主要挑战是纳米颗粒被肝脏和单核吞噬系统非特异性摄取，部分原因是内源性血清蛋白在LNP表面的吸附。可调的LNP表面化学性质可以使其有效地传递到各种器官和细胞类型。在这里，我们描述了一种静电吸附生物活性聚电解质到LNPs上以创建层状LNPs （LLNPs）的方法。LNP核的核酸载货量和组成脂质各不相同，稳定地由四种生物相关的聚阴离子构成：透明质酸（HA）、聚l -天冬氨酸（PLD）、聚l -谷氨酸（PLE）和聚丙烯酸酯（PAA）。我们进一步研究了四种表面多阴离子对细胞培养中mRNA和pdna负载LNPs的转染和摄取的影响。PLD-和PLE-LLNPs在免疫细胞中的mRNA转染量比未分层LNPs增加两倍。HA-LLNPs使上皮细胞和免疫细胞的pDNA转染率增加了两倍以上。在健康的C57BL/6小鼠模型中，肝脏和脾脏中PLE-和HA-LLNPs的转染量比未分层的LNPs增加1.8至2.5倍。这些结果表明，LbL组装是一个通用的、高度可调的平台，可以改变LNPs的靶向特异性、稳定性和转染效率，并将其他带电靶向和治疗分子纳入这些系统。
Rapid advances in nucleic acid therapies highlight the immense therapeutic potential of genetic therapeutics. Lipid nanoparticles (LNPs) are highly potent nonviral transfection agents that can encapsulate and deliver various nucleic acid therapeutics, including but not limited to messenger RNA (mRNA), silencing RNA (siRNA), and plasmid DNA (pDNA). However, a major challenge of targeted LNP-mediated systemic delivery is the nanoparticles’ nonspecific uptake by the liver and the mononuclear phagocytic system, due partly to the adsorption of endogenous serum proteins onto LNP surfaces. Tunable LNP surface chemistries may enable efficacious delivery across a range of organs and cell types. Here, we describe a method to electrostatically adsorb bioactive polyelectrolytes onto LNPs to create layered LNPs (LLNPs). LNP cores varying in nucleic acid cargo and component lipids were stably layered with four biologically relevant polyanions: hyaluronate (HA), poly-L-aspartate (PLD), poly-L-glutamate (PLE), and polyacrylate (PAA). We further investigated the impact of the four surface polyanions on the transfection and uptake of mRNA- and pDNA-loaded LNPs in cell cultures. PLD- and PLE-LLNPs increased mRNA transfection twofold over unlayered LNPs in immune cells. HA-LLNPs increased pDNA transfection rates by more than twofold in epithelial and immune cells. In a healthy C57BL/6 murine model, PLE- and HA-LLNPs increased transfection by 1.8-fold to 2.5-fold over unlayered LNPs in the liver and spleen. These results suggest that LbL assembly is a generalizable, highly tunable platform to modify the targeting specificity, stability, and transfection efficacy of LNPs, as well as incorporate other charged targeting and therapeutic molecules into these systems.