- 作者: Sin-Yu Chen, Yi-Ling Chen, Po-Chen Li, Tai-Shan Cheng, Yeh-Shiu Chu, Yi-Shan Shen, Hsin-Tung Chen, Wei-Ni Tsai, Chien-Ling Huang, Martin Sieber, Yuan-Chieh Yeh, Hsiao-Sheng Liu, Chi-Ling Chiang, Chih-Hung Chang, Andrew S. Lee, Yen-Han Tseng, Ly James Lee, Hsiu-Jung Liao, Hon-Kan Yip & Chi-Ying F. Huang
- 作者服務機構: 1.Bionet Therapeutics, Taipei, 114065, Taiwan 2.Brain Research Center, National Yang Ming Chiao Tung University, Taipei, 112304, Taiwan 3.Center for Cancer Research, College of Medicine, Kaohsiung Medical University, Kaohsiung, 807378, Taiwan 4.Center for Shockwave Medicine and Tissue Engineering, Kaohsiung Chang Gung Memorial Hospital, Kaohsiung, 833401, Taiwan 5.Comprehensive Cancer Center, College of Medicine, The Ohio State University, Columbus, OH, 43210, USA 6.Department of Biochemistry, School of Medicine, Kaohsiung Medical University, Kaohsiung, 807378, Taiwan 7.Department of Biomedical Engineering, National Taiwan University, Taipei, 106319, Taiwan 8.Department of Chemical and Biomolecular Engineering, The Ohio State University, Columbus, OH, 43210, USA 9.Department of Chest Medicine, Taipei Veterans General Hospital, Taipei, 112201, Taiwan 10.Department of Medical Research, China Medical University Hospital, China Medical University, Taichung, 404328, Taiwan 11.Department of Medical Research, Far Eastern Memorial Hospital, New Taipei City, 220216, Taiwan 12.Department of Microbiology and Immunology, College of Medicine, National Cheng Kung University, Tainan, 701401, Taiwan 13.Department of Nursing, Asia University, Taichung, 413305, Taiwan 14.Department of Orthopedic Surgery, Far Eastern Memorial Hospital, New Taipei City, 220216, Taiwan 15.Department of Traditional Chinese Medicine, Chang Gung Memorial Hospital, Keelung, 204201, Taiwan 16.Division of Cardiology, Department of Internal Medicine, Kaohsiung Chang Gung Memorial Hospital and Chang Gung University College of Medicine, Kaohsiung, 833401, Taiwan 17.Graduate School of Biotechnology and Bioengineering, Yuan Ze University, Taoyuan, 320315, Taiwan 18.Institute for Translational Research in Biomedicine, Kaohsiung Chang Gung Memorial Hospital, Kaohsiung, 833401, Taiwan 19.Institute of Biopharmaceutical Sciences, College of Pharmaceutical Sciences, National Yang Ming Chiao Tung University, Taipei, 112304, Taiwan 20.Program in Molecular Medicine, College of Life Sciences, National Yang Ming Chiao Tung University, Taipei, 112304, Taiwan 21.Spot Biosystems Ltd., Palo Alto, CA, 94305, USA 22.Teaching and Research Center, Kaohsiung Municipal Siaogang Hospital, Kaohsiung Medical University Hospital, Kaohsiung Medical University, Kaohsiung, 812015, Taiwan
- 中文摘要:
- 英文摘要:
Background
Acute lung injury (ALI) is a life-threatening respiratory condition characterized by severe inflammation and lung tissue damage, frequently causing rapid respiratory failure and long-term complications. The microRNA let-7a-5p is involved in the progression of lung injury, inflammation, and fibrosis by regulating immune cell activation and cytokine production. This study aims to use an innovative cellular electroporation platform to generate extracellular vesicles (EVs) carring let-7a-5p (EV-let-7a-5p) derived from transfected Wharton’s jelly-mesenchymal stem cells (WJ-MSCs) as a potential gene therapy for ALI.
Methods
A cellular nanoporation (CNP) method was used to induce the production and release of EV-let-7a-5p from WJ-MSCs transfected with the relevant plasmid DNA. EV-let-7a-5p in the conditioned medium were isolated using a tangential flow filtration (TFF) system. EV characterization followed the minimal consensus guidelines outlined by the International Society for Extracellular Vesicles. We conducted a thorough set of therapeutic assessments, including the antifibrotic effects using a transforming growth factor beta (TGF-β)-induced cell model, the modulation effects on macrophage polarization, and the influence of EV-let-7a-5p in a rat model of hyperoxia-induced ALI.
Results
The CNP platform significantly increased EV secretion from transfected WJ-MSCs, and the encapsulated let-7a-5p in engineered EVs was markedly higher than that in untreated WJ-MSCs. These EV-let-7a-5p did not influence cell proliferation and effectively mitigated the TGF-β-induced fibrotic phenotype by downregulating SMAD2/3 phosphorylation in LL29 cells. Furthermore, EV-let-7a-5p regulated M2-like macrophage activation in an inflammatory microenvironment and significantly induced interleukin (IL)-10 secretion, demonstrating their modulatory effect on inflammation. Administering EVs from untreated WJ-MSCs slightly improved lung function and increased let-7a-5p expression in plasma in the hyperoxia-induced ALI rat model. In comparison, EV-let-7a-5p significantly reduced macrophage infiltration and collagen deposition while increasing IL-10 expression, causing a substantial improvement in lung function.
Conclusion
This study reveals that the use of the CNP platform to stimulate and transfect WJ-MSCs could generate an abundance of let-7a-5p-enriched EVs, which underscores the therapeutic potential in countering inflammatory responses, fibrotic activation, and hyperoxia-induced lung injury. These results provide potential avenues for developing innovative therapeutic approaches for more effective interventions in ALI. - 中文關鍵字:
- 英文關鍵字: Let-7a-5p, Extracellular vesicles, Mesenchymal stem cells, Cellular nanoporation, Anti-infammation, Acute lung injury