将信使核糖核酸(mRNA)与含有轻度阳离子可电离脂质的预形成脂质纳米颗粒混合,可实现高效的体外和体内mRNA转染。

Admixing of mRNA with Pre-Formed Lipid Nanoparticles Containing a Slightly-Cationic Ionizable Lipid Allows for Efficient mRNA Transfection In Vitro and In Vivo.

作者信息

Wang Haixiu, Lauwers Heleen, Gontsarik Mark, Chen Yong, Sanders Niek N, Cheng Miffy Hok Yan, De Geest Bruno G, Zhong Zifu

机构信息

Department of Pharmaceutics, Ghent University, Ottergemsesteenweg 460, Gent, 9000, Belgium.

Laboratory of Gene Therapy, Ghent University, Heidestraat 19, Merelbeke, 9820, Belgium.

出版信息

Adv Healthc Mater. 2025 Jun 25:e2501788. doi: 10.1002/adhm.202501788.

Abstract

Therapeutic mRNA has emerged as a powerful tool in medicine. However, due to its fragility and large size, mRNA requires a carrier for delivery into the cellular cytosol. Lipid nanoparticles (LNPs), produced by rapidly mixing an aqueous mRNA solution with an ethanolic solution containing lipids, are currently considered the most advanced carriers for this purpose. Electrostatic interactions between mRNA and the ionizable cationic lipid, combined with hydrophobic interactions among all lipids, lead to self-assembly into LNPs that accommodate the mRNA in their core. In this study, whether mixing mRNA with pre-formed, empty LNPs (eLNPs) in an aqueous medium can be a viable alternative for mRNA expression is investigated. It is confirmed that mRNA can associate with eLNPs via electrostatic interactions, with the effectiveness of this association depending on the surface charge of the eLNPs and the ionizable lipid component. Furthermore, post-loading mRNA into eLNPs demonstrates mRNA expression levels comparable to conventional LNP(mRNA) formulations, both in vitro and in mice. This method of leveraging eLNPs offers a practical alternative to conventional LNP(mRNA) formulation for the rapid screening of multiple mRNAs. It can also enable straightforward use of LNPs for mRNA transfection by users who do not have the capacity to perform LNP formulation.

摘要

治疗性mRNA已成为医学领域的一种强大工具。然而,由于其脆弱性和较大的尺寸,mRNA需要载体才能递送至细胞质溶胶中。通过将mRNA水溶液与含脂质的乙醇溶液快速混合制备的脂质纳米颗粒(LNP),目前被认为是用于此目的的最先进载体。mRNA与可电离阳离子脂质之间的静电相互作用,以及所有脂质之间的疏水相互作用,导致自组装形成将mRNA容纳在其核心的LNP。在本研究中,研究了在水性介质中将mRNA与预先形成的空LNP(eLNP)混合是否可以作为mRNA表达的可行替代方法。证实mRNA可通过静电相互作用与eLNP结合,这种结合的有效性取决于eLNP的表面电荷和可电离脂质成分。此外,将mRNA后装载到eLNP中在体外和小鼠体内均显示出与传统LNP(mRNA)制剂相当的mRNA表达水平。这种利用eLNP的方法为快速筛选多种mRNA提供了一种替代传统LNP(mRNA)制剂的实用方法。它还可以使没有能力进行LNP制剂制备的用户直接使用LNP进行mRNA转染。

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