Department of Chemistry, The University of Melbourne, Parkville, Victoria 3010, Australia.
Monash Institute of Pharmaceutical Sciences, Monash University, Parkville, Victoria 3010, Australia.
ACS Appl Mater Interfaces. 2022 Jan 26;14(3):3653-3661. doi: 10.1021/acsami.1c18359. Epub 2021 Dec 29.
All nanoparticles have the potential to revolutionize the delivery of therapeutic cargo such as peptides, proteins, and RNA. However, effective cytosolic delivery of cargo from nanoparticles represents a significant challenge in the design of more efficient drug delivery vehicles. Recently, research has centered on designing nanoparticles with the capacity to escape endosomes by responding to biological stimuli such as changes in pH, which occur when nanoparticles are internalized into the endo-/lysosomal pathway. Current endosomal escape assays rely on indirect measurements and yield little quantitative information, which hinders the design of more efficient drug delivery vehicles. Therefore, we adapted the highly sensitive split luciferase endosomal escape quantification (SLEEQ) assay to better understand nanoparticle-induced endosomal escape. We applied SLEEQ to evaluate the endosomal escape behavior of two pH-responsive nanoparticles: the first with a poly(2-diisopropylamino ethyl methacrylate) (PDPAEMA) core and the second with 1:1 ratio of poly(2-diethylamino ethyl methacrylate) (PDEAEMA) and PDPAEMA. SLEEQ directly measured the cytosolic delivery and showed that engineering the nanoparticle disassembly pH could improve the endosomal escape efficiency by fivefold. SLEEQ is a versatile assay that can be used for a wide range of nanomaterials and will improve the development of drug delivery vehicles in the future.
所有的纳米颗粒都有可能彻底改变治疗性货物（如肽、蛋白质和 RNA）的输送方式。然而，将货物从纳米颗粒有效递送至细胞质中，这在设计更高效的药物输送载体方面是一个重大挑战。最近，研究的重点是设计能够响应生物刺激（如纳米颗粒被内吞进入内体/溶酶体途径时发生的 pH 变化）从而逃离内体的纳米颗粒。目前的内体逃逸检测依赖于间接测量，并且只能提供很少的定量信息，这阻碍了更高效的药物输送载体的设计。因此，我们采用了高度灵敏的分裂荧光素酶内体逃逸定量（SLEEQ）检测来更好地理解纳米颗粒诱导的内体逃逸。我们应用 SLEEQ 来评估两种 pH 响应纳米颗粒的内体逃逸行为：第一种是具有聚（2-二异丙基氨基乙基甲基丙烯酸酯）（PDPAEMA）核的纳米颗粒，第二种是聚（2-二乙基氨基乙基甲基丙烯酸酯）（PDEAEMA）和 PDPAEMA 比例为 1:1 的纳米颗粒。SLEEQ 直接测量了细胞质中的递送情况，并表明通过工程设计纳米颗粒的组装 pH 可以将内体逃逸效率提高五倍。SLEEQ 是一种多功能的检测方法，可用于广泛的纳米材料，并且将在未来改善药物输送载体的开发。
