3D hESC exosomes enriched with miR-6766-3p ameliorates liver fibrosis by attenuating activated stellate cells through targeting the TGFβRII-SMADS pathway
| Autor: | Chuxin Chen, Zhiyong Zhong, Qicong Chen, Weili Gu, Haibin Wu, Shuai Zhang, Xiajing Li, Honglin Chen, Yaqi Qiu, Bailin Wang, Shoupei Liu, Yimeng Ou, Yingjie Fu, Ning Wang, Mark A Zern, Keqing Ma, Yuyou Duan, Jie Cao, Xianglian Tang, Jinsong Li, Tingting Guo |
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| Jazyk: | angličtina |
| Rok vydání: | 2021 |
| Předmět: |
Liver Cirrhosis
Male Liver fibrosis Human Embryonic Stem Cells Biomedical Engineering Pharmaceutical Science Medicine (miscellaneous) Bioengineering Smad Proteins Exosomes Applied Microbiology and Biotechnology Collagen Type I Mice Transforming Growth Factor beta Medical technology Animals Humans Cell Culture Techniques Three Dimensional R855-855.5 RNA Small Interfering Cell Proliferation Mice Inbred ICR Chemistry Cell spheroids Research TGFβRII-SMADS pathway Receptor Transforming Growth Factor-beta Type II Antagomirs Microvesicles microRNAs Disease Models Animal Hepatic stellate cell Cancer research Molecular Medicine RNA Interference TP248.13-248.65 Biotechnology Signal Transduction |
| Zdroj: | Journal of Nanobiotechnology Journal of Nanobiotechnology, Vol 19, Iss 1, Pp 1-26 (2021) |
| ISSN: | 1477-3155 |
| Popis: | Background Exosomes secreted from stem cells exerted salutary effects on the fibrotic liver. Herein, the roles of exosomes derived from human embryonic stem cell (hESC) in anti-fibrosis were extensively investigated. Compared with two-dimensional (2D) culture, the clinical and biological relevance of three-dimensional (3D) cell spheroids were greater because of their higher regeneration potential since they behave more like cells in vivo. In our study, exosomes derived from 3D human embryonic stem cells (hESC) spheroids and the monolayer (2D) hESCs were collected and compared the therapeutic potential for fibrotic liver in vitro and in vivo. Results In vitro, PKH26 labeled-hESC-Exosomes were shown to be internalized and integrated into TGFβ-activated-LX2 cells, and reduced the expression of profibrogenic markers, thereby regulating cellular phenotypes. TPEF imaging indicated that PKH26-labeled-3D-hESC-Exsomes possessed an enhanced capacity to accumulate in the livers and exhibited more dramatic therapeutic potential in the injured livers of fibrosis mouse model. 3D-hESC-Exosomes decreased profibrogenic markers and liver injury markers, and improved the level of liver functioning proteins, eventually restoring liver function of fibrosis mice. miRNA array revealed a significant enrichment of miR-6766-3p in 3D-hESC-Exosomes, moreover, bioinformatics and dual luciferase reporter assay identified and confirmed the TGFβRII gene as the target of miR-6766-3p. Furthermore, the delivery of miR-6766-3p into activated-LX2 cells decreased cell proliferation, chemotaxis and profibrotic effects, and further investigation demonstrated that the expression of target gene TGFβRII and its downstream SMADs proteins, especially phosphorylated protein p-SMAD2/3 was also notably down-regulated by miR-6766-3p. These findings unveiled that miR-6766-3p in 3D-hESC-Exosomes inactivated SMADs signaling by inhibiting TGFβRII expression, consequently attenuating stellate cell activation and suppressing liver fibrosis. Conclusions Our results showed that miR-6766-3p in the 3D-hESC-Exosomes inactivates smads signaling by restraining TGFβRII expression, attenuated LX2 cell activation and suppressed liver fibrosis, suggesting that 3D-hESC-Exosome enriched-miR-6766-3p is a novel anti-fibrotic therapeutics for treating chronic liver disease. These results also proposed a significant strategy that 3D-Exo could be used as natural nanoparticles to rescue liver injury via delivering antifibrotic miR-6766-3p. Graphical Abstract |
| Databáze: | OpenAIRE |
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