van Dijk Fransien, Hazelhoff Christa M, Post Eduard, Prins Gerian G H, Rombouts Krista, Poelstra Klaas, Olinga Peter, Beljaars Leonie
Groningen Research Institute of Pharmacy, Department of Pharmacokinetics, Toxicology and Targeting, University of Groningen, 9713 AV Groningen, The Netherlands.
Groningen Research Institute of Pharmacy, Department of Pharmaceutical Technology and Biopharmacy, University of Groningen, 9713 AV Groningen, The Netherlands.
Pharmaceutics. 2020 Mar 20;12(3):278. doi: 10.3390/pharmaceutics12030278.
The pivotal cell involved in the pathogenesis of liver fibrosis, i.e., the activated hepatic stellate cell (HSC), has a wide range of activities during the initiation, progression and even regression of the disease. These HSC-related activities encompass cellular activation, matrix synthesis and degradation, proliferation, contraction, chemotaxis and inflammatory signaling. When determining the in vitro and in vivo effectivity of novel antifibrotic therapies, the readout is currently mainly based on gene and protein levels of α-smooth muscle actin (α-SMA) and the fibrillar collagens (type I and III). We advocate for a more comprehensive approach in addition to these markers when screening potential antifibrotic drugs that interfere with HSCs. Therefore, we aimed to develop a gene panel for human in vitro and ex vivo drug screening models, addressing each of the HSC-activities with at least one gene, comprising, in total, 16 genes. We determined the gene expression in various human stellate cells, ranging from primary cells to cell lines with an HSC-origin, and human liver slices and stimulated them with two key profibrotic factors, i.e., transforming growth factor β (TGFβ) or platelet-derived growth factor BB (PDGF-BB). We demonstrated that freshly isolated HSCs showed the strongest and highest variety of responses to these profibrotic stimuli, in particular following PDGF-BB stimulation, while cell lines were limited in their responses. Moreover, we verified these gene expression profiles in human precision-cut liver slices and showed similarities with the TGFβ- and PDGF-BB-related fibrotic responses, as observed in the primary HSCs. With this study, we encourage researchers to get off the beaten track when testing antifibrotic compounds by including more HSC-related markers in their future work. This way, potential compounds will be screened more extensively, which might increase the likelihood of developing effective antifibrotic drugs.
肝纤维化发病机制中的关键细胞，即活化的肝星状细胞（HSC），在疾病的起始、进展甚至消退过程中都有广泛的活动。这些与HSC相关的活动包括细胞活化、基质合成与降解、增殖、收缩、趋化性和炎症信号传导。在确定新型抗纤维化疗法的体外和体内有效性时，目前的检测指标主要基于α-平滑肌肌动蛋白（α-SMA）和纤维状胶原蛋白（I型和III型）的基因和蛋白水平。我们主张在筛选干扰HSC的潜在抗纤维化药物时，除了这些标志物外，采用更全面的方法。因此，我们旨在开发一个用于人类体外和离体药物筛选模型的基因panel，用至少一个基因来涵盖HSC的每一项活动，总共包括16个基因。我们测定了各种人类星状细胞中的基因表达，这些细胞从原代细胞到具有HSC起源的细胞系，以及人类肝切片，并分别用两种关键的促纤维化因子，即转化生长因子β（TGFβ）或血小板衍生生长因子BB（PDGF-BB）进行刺激。我们证明，新鲜分离的HSC对这些促纤维化刺激表现出最强且最多样化的反应，特别是在PDGF-BB刺激后，而细胞系的反应则有限。此外，我们在人类精密肝切片中验证了这些基因表达谱，并显示出与原代HSC中观察到的TGFβ和PDGF-BB相关的纤维化反应相似。通过这项研究，我们鼓励研究人员在测试抗纤维化化合物时另辟蹊径，在未来的工作中纳入更多与HSC相关的标志物。这样，潜在的化合物将得到更广泛的筛选，这可能会增加开发有效抗纤维化药物的可能性。
