A Novel Dual-Reporter System Reveals Distinct Characteristics of Exosome-Mediated Protein Secretion in Human Cells

Biological Procedures Online

Research

Biao Lu¹  Pengyang Zhang¹  Christopher Olson¹  Joy Ku¹  Renceh Flojo¹  Darin Boyes² 

[1] Department of Bioengineering, Santa Clara University, 500 El Camino Real, 95053, Santa Clara, CA, USA;Department of Biology, Santa Clara University, 500 El Camino Real, 95053, Santa Clara, CA, USA;

关键词: Exosome;    Microvesicle;    Extracellular vesicle;    Dual-reporter system;    Cell-to-cell communication;    Bioluminescence luciferase;    Fluorescence protein;    Protein secretion;   

DOI  :  10.1186/s12575-023-00219-w

 received in 2023-06-29, accepted in 2023-09-05,  发布年份 2023

来源: Springer

【 摘 要 】

BackgroundExosomes, a special subtype of extracellular vesicles derived from human cells, serve as vital mediators of intercellular communication by transporting diverse bioactive cargos, including proteins and enzymes. However, the underlying mechanisms governing exosome secretion and regulation remain poorly understood. In this study, we employed a dual-reporter system consisting of bioluminescent Gaussia luciferase and fluorescent proteins to investigate the dynamics and regulation of exosome secretion in cultured human cells.ResultsOur results demonstrated that the engineered dual-reporters effectively monitored both exosome-mediated and ER-Golgi-mediated secretory pathways in a specific and quantitative manner. Notably, we observed distinct characteristics of exosome-mediated protein secretion, including significantly lower capacity and different dynamics compared to the ER-Golgi pathway. This phenomenon was observed in human kidney 293T cells and liver HepG2 cells, emphasizing the conserved nature of exosome-mediated secretion across cell types. Furthermore, we investigated the impact of brefeldin A (BFA), an inhibitor of ER-to-Golgi membrane trafficking, on protein secretion. Interestingly, BFA inhibited protein secretion via the ER-Golgi pathway while stimulating exosome-mediated protein secretion under same experimental conditions.ConclusionsCollectively, our study highlights the utility of the dual-reporter system for real-time monitoring and quantitative analysis of protein secretion through conventional ER-Golgi and unconventional exosome pathways. Moreover, our findings unveil distinct features of exosome-mediated protein secretion, shedding light on its differential capacity, dynamics, and regulatory mechanisms compared to ER-Golgi-mediated proteins in human cells.

【 授权许可】

CC BY   
© BioMed Central Ltd., part of Springer Nature 2023

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