Abstract
Liver fibrosis is a significant health burden, marked by the consistent deposition of collagen. Unfortunately, the currently available treatment approaches for this condition are far from optimal. Lysyl oxidase-like protein 2 (LOXL2) secreted by hepatic stellate cells (HSCs) is a crucial player in the cross-linking of matrix collagen and is a significant target for treating liver fibrosis. Mesenchymal stem cell-derived small extracellular vesicles (MSC-sEVs) have been proposed as a potential treatment option for chronic liver disorders. Previous studies have found that MSC-sEV can be used for microRNA delivery into target cells or tissues. It is currently unclear whether microRNA-4465 (miR-4465) can target LOXL2 and inhibit HSC activation. Additionally, it is uncertain whether MSC-sEV can be utilized as a gene therapy vector to carry miR-4465 and effectively inhibit the progression of liver fibrosis. This study explored the effect of miR-4465-modified MSC-sEV (MSC-sEVmiR-4465) on LOXL2 expression and liver fibrosis development. The results showed that miR-4465 can bind specifically to the promoter of the LOXL2 gene in HSC. Moreover, MSC-sEVmiR-4465 inhibited HSC activation and collagen expression by downregulating LOXL2 expression in vitro. MSC-sEVmiR-4465 injection could reduce HSC activation and collagen deposition in the CCl4-induced mouse model. MSC-sEVmiR-4465 mediating via LOXL2 also hindered the migration and invasion of HepG2 cells. In conclusion, we found that MSC-sEV can deliver miR-4465 into HSC to alleviate liver fibrosis via altering LOXL2, which might provide a promising therapeutic strategy for liver diseases.
摘要
肝纤维化是慢性肝病向肝硬化发展的关键病理过程, 其特征是胶原蛋白的过量沉积。然而, 目前对于肝纤维化仍缺乏有效的治疗方法。肝星状细胞(HSC)分泌的赖氨酰氧化酶样蛋白2(LOXL2)是胶原交联和HSC活化的关键分子, 也是治疗肝纤维化的重要靶点。研究表明, 间质干细胞来源的小细胞外囊泡(MSC-sEV)是治疗慢性肝病的有效方法。MSC-sEV可将miRNA递送到靶细胞或组织中。但目前尚不明确miR-4465是否能够靶向LOXL2抑制HSC活化, 同时MSC-sEV是否可以作为基因治疗载体通过携带miR-4465从而抑制肝纤维化进展也不清楚。本研究探讨了miR-4465对于LOXL2的调控作用及miR-4465修饰的MSC-sEV(MSC-sEVmiR-4465)对LOXL2表达和肝纤维化进展的影响。结果表明, miR-4465可以靶向HSC中LOXL2基因的启动子。此外, MSC-sEVmiR-4465在体外通过下调LOXL2的表达来抑制HSC的活化和胶原的表达。而MSC-sEVmiR-4465注射可减少CCl4诱导的小鼠模型中HSC的活化和胶原沉积。MSC-sEVmiR-4465也能通过调控LOXL2抑制肝癌细胞HepG2的迁移和侵袭。总之, 我们发现MSC-sEV可以通过递送miR-4465到HSC中靶向LOXL2延缓肝纤维化的进展, 有望成为未来肝病治疗的一种新策略。
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Data are available on request from the authors.
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Acknowledgments
This work was supported by the National Natural Science Foundation of China (No. 82272421), the Jiangsu Provincial Key Research and Development Program (No. BE2021690), the Changzhou’s 14th Five-year Plan Project to Train Highlevel Health Professionals (No. 2022CZLJ027), the Scientific Project of Jiangsu Health Commission (No. Z2020038), and the Changzhou Sci & Tech Program (No. CJ20220164), China.
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Yan** WANG, Yifei CHEN, and Fuji YANG contributed to investigation, validation, formal analysis, and writing–original draft. **aolong YU, Ying CHU, and **g ZHOU performed methodology and software. Jianbo XI and Yongmin YAN contributed to conceptualization, funding acquisition, and writing–review & editing. All authors have read and approved the final manuscript, and therefore, have full access to all the data in the study and take responsibility for the integrity and security of the data.
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Yan** WANG, Yifei CHEN, Fuji YANG, **aolong YU, Ying CHU, **g ZHOU, Yongmin YAN, and Jianbo XI declare that they have no conflict of interest.
All institutional and national guidelines for the care and use of laboratory animals were followed. The animal study protocol followed the ethical policies and procedures approved by the Jiangsu University Ethics Committee (Approval No. UJS-IACUC-AP-2020033127).
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Wang, Y., Chen, Y., Yang, F. et al. MiR-4465-modified mesenchymal stem cell-derived small extracellular vesicles inhibit liver fibrosis development via targeting LOXL2 expression. J. Zhejiang Univ. Sci. B (2024). https://doi.org/10.1631/jzus.B2300305
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DOI: https://doi.org/10.1631/jzus.B2300305
Key words
- Mesenchymal stem cell (MSC)
- Small extracellular vesicle (sEV)
- MicroRNA-4465 (miR-4465)
- Hepatic stellate cell (HSC)
- Liver fibrosis