Cell, Gene and RNA Therapy, Discovery Sciences, R&D, AstraZeneca, Cambridge, UK.
Advanced Drug Delivery, Pharmaceutical Sciences, R&D, AstraZeneca, Cambridge, UK.
Nucleic Acids Res. 2024 Sep 9;52(16):9384-9396. doi: 10.1093/nar/gkae648.
Lipid nanoparticles (LNPs) have been demonstrated to hold great promise for the clinical advancement of RNA therapeutics. Continued exploration of LNPs for application in new disease areas requires identification and optimization of leads in a high throughput way. Currently available high throughput in vivo screening platforms are well suited to screen for cellular uptake but less so for functional cargo delivery. We report on a platform which measures functional delivery of LNPs using unique peptide 'barcodes'. We describe the design and selection of the peptide barcodes and the evaluation of these for the screening of LNPs. We show that proteomic analysis of peptide barcodes correlates with quantification and efficacy of barcoded reporter proteins both in vitro and in vivo and, that the ranking of selected LNPs using peptide barcodes in a pool correlates with ranking using alternative methods in groups of animals treated with individual LNPs. We show that this system is sensitive, selective, and capable of reducing the size of an in vivo study by screening up to 10 unique formulations in a single pool, thus accelerating the discovery of new technologies for mRNA delivery.
脂质纳米颗粒（LNPs）在 RNA 治疗学的临床进展中具有巨大的应用前景。为了将 LNPs 应用于新的疾病领域，需要以高通量的方式鉴定和优化先导化合物。目前可用的高通量体内筛选平台非常适合筛选细胞摄取，但在功能性货物递送上效果较差。我们报告了一种使用独特肽“条形码”测量 LNP 功能递药的平台。我们描述了肽条形码的设计和选择，以及对这些条形码进行筛选的评估。我们表明，肽条形码的蛋白质组学分析与体外和体内报告蛋白条形码的定量和功效相关，并且使用肽条形码对池中的选定 LNPs 进行排序与使用单独用 LNPs 处理的动物组中的替代方法进行排序相关。我们表明，该系统具有敏感性、选择性，并且能够通过在单个池中筛选多达 10 种独特的制剂来减少体内研究的规模，从而加速发现新的 mRNA 递送技术。
