Issue

Vol. 16 (2024)

Molecular Mechanisms of Intracellular Delivery of Nanoparticles Monitored by an Enzyme-Induced Proximity Labeling

https://doi.org/10.1007/s40820-023-01313-0
Junji Ren
Department of Pharmaceutics School of Pharmaceutical Sciences, Peking University, 38 Xueyuan Rd, Haidian District, Beijing 100191, People’s Republic of China
Zibin Zhang
Department of Pharmaceutics School of Pharmaceutical Sciences, Peking University, 38 Xueyuan Rd, Haidian District, Beijing 100191, People’s Republic of China
Shuo Geng
Department of Pharmaceutics School of Pharmaceutical Sciences, Peking University, 38 Xueyuan Rd, Haidian District, Beijing 100191, People’s Republic of China
Yuxi Cheng
Department of Pharmaceutics School of Pharmaceutical Sciences, Peking University, 38 Xueyuan Rd, Haidian District, Beijing 100191, People’s Republic of China
Huize Han
Department of Pharmaceutics School of Pharmaceutical Sciences, Peking University, 38 Xueyuan Rd, Haidian District, Beijing 100191, People’s Republic of China
Zhipu Fan
Department of Pharmaceutics School of Pharmaceutical Sciences, Peking University, 38 Xueyuan Rd, Haidian District, Beijing 100191, People’s Republic of China
Wenbing Dai
Department of Pharmaceutics School of Pharmaceutical Sciences, Peking University, 38 Xueyuan Rd, Haidian District, Beijing 100191, People’s Republic of China
Hua Zhang
Department of Pharmaceutics School of Pharmaceutical Sciences, Peking University, 38 Xueyuan Rd, Haidian District, Beijing 100191, People’s Republic of China
Xueqing Wang
Department of Pharmaceutics School of Pharmaceutical Sciences, Peking University, 38 Xueyuan Rd, Haidian District, Beijing 100191, People’s Republic of China
Qiang Zhang
Department of Pharmaceutics School of Pharmaceutical Sciences, Peking University, 38 Xueyuan Rd, Haidian District, Beijing 100191, People’s Republic of China
Bing He
Department of Pharmaceutics School of Pharmaceutical Sciences, Peking University, 38 Xueyuan Rd, Haidian District, Beijing 100191, People’s Republic of China

Corresponding Author: Bing He

hebingmumu@bjmu.edu.cn

Nano-Micro Letters, Vol. 16 (2024), Article Number: 103

Abstract

Achieving increasingly finely targeted drug delivery to organs, tissues, cells, and even to intracellular biomacromolecules is one of the core goals of nanomedicines. As the delivery destination is refined to cellular and subcellular targets, it is essential to explore the delivery of nanomedicines at the molecular level. However, due to the lack of technical methods, the molecular mechanism of the intracellular delivery of nanomedicines remains unclear to date. Here, we develop an enzyme-induced proximity labeling technology in nanoparticles (nano-EPL) for the real-time monitoring of proteins that interact with intracellular nanomedicines. Poly(lactic-co-glycolic acid) nanoparticles coupled with horseradish peroxidase (HRP) were fabricated as a model (HRP(+)-PNPs) to evaluate the molecular mechanism of nano delivery in macrophages. By adding the labeling probe biotin-phenol and the catalytic substrate H2O2 at different time points in cellular delivery, nano-EPL technology was validated for the real-time in situ labeling of proteins interacting with nanoparticles. Nano-EPL achieves the dynamic molecular profiling of 740 proteins to map the intracellular delivery of HRP (+)-PNPs in macrophages over time. Based on dynamic clustering analysis of these proteins, we further discovered that different organelles, including endosomes, lysosomes, the endoplasmic reticulum, and the Golgi apparatus, are involved in delivery with distinct participation timelines. More importantly, the engagement of these organelles differentially affects the drug delivery efficiency, reflecting the spatial–temporal heterogeneity of nano delivery in cells. In summary, these findings highlight a significant methodological advance toward understanding the molecular mechanisms involved in the intracellular delivery of nanomedicines.


Highlights:
1 Novel enzyme-induced proximity labeling technology in nanoparticles (nano-EPL).
2 Nano-EPL enables dynamic molecular mapping of the intracellular delivery of nanoparticles in macrophages.
3 Nano-EPL enables the elucidation of a comprehensive phagosome-centered timeline during the vesicular transportation of nanoparticles, revealing distinct organelle engagement and its differential impact on drug delivery efficiency.

Keywords

Enzyme-induced proximity labeling Intracellular delivery Nano-protein interaction Dynamic molecule profiling Macrophages
Ren, Junji, Zibin Zhang, Shuo Geng, Yuxi Cheng, Huize Han, Zhipu Fan, Wenbing Dai, Hua Zhang, Xueqing Wang, Qiang Zhang, and Bing He. 2024. “Molecular Mechanisms of Intracellular Delivery of Nanoparticles Monitored by an Enzyme-Induced Proximity Labeling”. Nano-Micro Letters 16 (February):103. https://doi.org/10.1007/s40820-023-01313-0.
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