Institute of Pharmacy, Friedrich Schiller University of Jena, Jena, Germany.
Department of Chemistry, Biochemistry and Pharmaceutical Sciences, University of Bern, Bern, Switzerland.
Commun Biol. 2022 Oct 30;5(1):1155. doi: 10.1038/s42003-022-04123-z.
Uncovering the complex cellular mechanisms underlying hepatic fibrogenesis could expedite the development of effective treatments and noninvasive diagnosis for liver fibrosis. The biochemical complexity of extracellular vesicles (EVs) and their role in intercellular communication make them an attractive tool to look for biomarkers as potential alternative to liver biopsies. We developed a solid set of methods to isolate and characterize EVs from differently treated human hepatic stellate cell (HSC) line LX-2, and we investigated their biological effect onto naïve LX-2, proving that EVs do play an active role in fibrogenesis. We mined our proteomic data for EV-associated proteins whose expression correlated with HSC treatment, choosing the matricellular protein SPARC as proof-of-concept for the feasibility of fluorescence nanoparticle-tracking analysis to determine an EV-based HSCs' fibrogenic phenotype. We thus used EVs to directly evaluate the efficacy of treatment with S80, a polyenylphosphatidylcholines-rich lipid, finding that S80 reduces the relative presence of SPARC-positive EVs. Here we correlated the cellular response to lipid-based antifibrotic treatment to the relative presence of a candidate protein marker associated with the released EVs. Along with providing insights into polyenylphosphatidylcholines treatments, our findings pave the way for precise and less invasive diagnostic analyses of hepatic fibrogenesis.
揭示肝纤维化的复杂细胞机制可以加速有效的治疗方法和非侵入性诊断肝纤维化的发展。细胞外囊泡（EVs）的生化复杂性及其在细胞间通讯中的作用使它们成为寻找生物标志物的有吸引力的工具，作为肝活检的潜在替代方法。我们开发了一套从不同处理的人肝星状细胞（HSC）系 LX-2 中分离和表征 EVs 的方法，并研究了它们对幼稚 LX-2 的生物学效应，证明 EVs 在纤维化中确实发挥了积极作用。我们对 EV 相关蛋白的蛋白质组学数据进行了挖掘，这些蛋白的表达与 HSC 治疗相关，选择细胞外基质蛋白 SPARC 作为荧光纳米颗粒跟踪分析确定基于 EV 的 HSCs 纤维化表型的可行性的概念验证。因此，我们使用 EVs 直接评估富含多烯磷脂酰胆碱的 S80 治疗的疗效，发现 S80 降低了 SPARC 阳性 EVs 的相对存在。在这里，我们将细胞对基于脂质的抗纤维化治疗的反应与与释放的 EV 相关的候选蛋白标记物的相对存在相关联。我们的研究结果不仅为多烯磷脂酰胆碱治疗提供了新的见解，还为肝纤维化的精确和非侵入性诊断分析铺平了道路。
