Department of Chemistry, University of Michigan, Ann Arbor, Michigan 48109, United States.
J Am Chem Soc. 2024 Nov 20;146(46):31885-31891. doi: 10.1021/jacs.4c11066. Epub 2024 Nov 7.
Ribonucleic acids (RNAs) are becoming increasingly significant in our search for improved biotherapeutics. RNA-based treatments offer high specificity, targeted delivery, and potentially lower-cost options for various debilitating human diseases. Despite these benefits, there are still relatively few FDA-approved RNA-based therapies, with the notable exceptions being the mRNA (mRNA) COVID-19 vaccines, which are delivered using lipid nanoparticle (LNP) systems. LNPs are distinctive drug delivery systems (DDSs) because of their ability to target specific cells, their biocompatibility, and their efficiency in merging with cellular membranes to enhance treatment effectiveness. While the biophysical landscapes of RNA structures in solution are relatively well understood, the impact of the LNP environment on RNA remains less clear. This study uses native ion mobility-mass spectrometry (IM-MS) and collision-induced unfolding (CIU) techniques to investigate how LNP encapsulation affects RNA structure and stability. We examine how various factors, such as ionization polarity, cofactor binding, lipid types, and lipid ratios, influence LNP-released RNA cargo. Our findings reveal that LNP DDSs induce significant changes in the structures and stabilities of their RNA cargo. However, the extent of these changes strongly depends on the type and composition of the lipids used. We conclude by discussing how IM-MS and CIU can aid in the continued development of more efficient LNP DDSs and improve DDS selection methodologies overall.
核糖核酸（RNAs）在我们寻找改进的生物疗法方面变得越来越重要。基于 RNA 的治疗方法具有高度的特异性、靶向递送，并且为各种使人衰弱的人类疾病提供了潜在的低成本选择。尽管有这些好处，但获得 FDA 批准的基于 RNA 的治疗方法仍然相对较少，值得注意的例外是使用脂质纳米颗粒（LNP）系统递送的 mRNA（mRNA）COVID-19 疫苗。LNP 是独特的药物递送系统（DDS），因为它们能够靶向特定细胞，具有生物相容性，并且能够与细胞膜融合以提高治疗效果。虽然 RNA 结构在溶液中的生物物理景观相对较好理解，但 LNP 环境对 RNA 的影响仍不明确。本研究使用天然离子淌度-质谱（IM-MS）和碰撞诱导解折叠（CIU）技术研究 LNP 封装如何影响 RNA 结构和稳定性。我们研究了各种因素，如电离极性、辅助因子结合、脂质类型和脂质比例，如何影响 LNP 释放的 RNA 货物。我们的研究结果表明，LNP DDS 会对其 RNA 货物的结构和稳定性产生显著影响。然而，这些变化的程度强烈取决于所用脂质的类型和组成。我们最后讨论了 IM-MS 和 CIU 如何帮助开发更有效的 LNP DDS，并总体上改进 DDS 选择方法。
