Our knowledge of complex pathological mechanisms underlying organ fibrosis is predominantly derived from animal studies. However, relevance of animal models for human disease is limited; therefore, an ex vivo model of human precision-cut tissue slices (PCTS) might become an indispensable tool in fibrosis research and drug development by bridging the animal–human translational gap. This study, presented as two parts, provides comprehensive characterization of the dynamic transcriptional changes in PCTS during culture by RNA sequencing. Part I investigates the differences in culture-induced responses in murine and human PCTS derived from healthy liver, kidney and gut. Part II delineates the molecular processes in cultured human PCTS generated from diseased liver, kidney and ileum. We demonstrated that culture was associated with extensive transcriptional changes and impacted PCTS in a universal way across the organs and two species by triggering an inflammatory response and fibrosis-related extracellular matrix (ECM) remodelling. All PCTS shared mRNA upregulation of IL-11 and ECM-degrading enzymes MMP3 and MMP10. Slice preparation and culturing activated numerous pathways across all PCTS, especially those involved in inflammation (IL-6, IL-8 and HMGB1 signalling) and tissue remodelling (osteoarthritis pathway and integrin signalling). Despite the converging effects of culture, PCTS display species-, organ- and pathology-specific differences in the regulation of genes and canonical pathways. The underlying pathology in human diseased PCTS endures and influences biological processes like cytokine release. Our study reinforces the use of PCTS as an ex vivo fibrosis model and supports future studies towards its validation as a preclinical tool for drug development.

我们对器官纤维化潜在的复杂病理机制的了解主要来自动物研究。但是，动物模型与人类疾病的相关性是有限的。因此，通过弥合动物与人类之间的翻译鸿沟，人类精密切割组织切片（PCTS）的体外模型可能成为纤维化研究和药物开发中必不可少的工具。这项研究分为两个部分，通过RNA测序对PCTS中动态转录变化的全面表征。第一部分研究了鼠类和人PCTS中源自健康肝脏，肾脏和肠道的培养物诱导反应的差异。第二部分描述了从患病的肝脏，肾脏和回肠产生的人类PCTS培养过程中的分子过程。我们证明了文化与广泛的转录变化有关，并通过触发炎症反应和与纤维化相关的细胞外基质（ECM）重塑，以跨器官和两个物种的通用方式影响了PCTS。所有PCTS都共享IL-11和ECM降解酶MMP3和MMP10的mRNA上调。切片的制备和培养激活了所有PCTS上的许多途径，尤其是涉及炎症（IL-6，IL-8和HMGB1信号传导）和组织重塑（骨关节炎途径和整联蛋白信号传导）的途径。尽管文化具有聚合作用，但PCTS在基因和规范途径的调节方面仍显示出物种，器官和病理学方面的差异。人类患PCTS的潜在病理学可以持久并影响诸如细胞因子释放的生物学过程。