Newcastle Fibrosis Research Group, Biosciences Institute, Faculty of Medical Sciences, Newcastle University, Newcastle upon Tyne, U.K.
FibroFind Ltd, FibroFind Laboratories, Medical School, Newcastle University, U.K.
Biosci Rep. 2024 Jun 26;44(6). doi: 10.1042/BSR20232147.
Liver fibrosis is the excessive accumulation of extracellular matrix proteins that occurs in most types of chronic liver disease. At the cellular level, liver fibrosis is associated with the activation of hepatic stellate cells (HSCs) which transdifferentiate into a myofibroblast-like phenotype that is contractile, proliferative and profibrogenic. HSC transdifferentiation induces genome-wide changes in gene expression that enable the cell to adopt its profibrogenic functions. We have previously identified that the deubiquitinase ubiquitin C-terminal hydrolase 1 (UCHL1) is highly induced following HSC activation; however, the cellular targets of its deubiquitinating activity are poorly defined. Here, we describe a role for UCHL1 in regulating the levels and activity of hypoxia-inducible factor 1 (HIF1), an oxygen-sensitive transcription factor, during HSC activation and liver fibrosis. HIF1 is elevated during HSC activation and promotes the expression of profibrotic mediator HIF target genes. Increased HIF1α expression correlated with induction of UCHL1 mRNA and protein with HSC activation. Genetic deletion or chemical inhibition of UCHL1 impaired HIF activity through reduction of HIF1α levels. Furthermore, our mechanistic studies have shown that UCHL1 elevates HIF activity through specific cleavage of degradative ubiquitin chains, elevates levels of pro-fibrotic gene expression and increases proliferation rates. As we also show that UCHL1 inhibition blunts fibrogenesis in a pre-clinical 3D human liver slice model of fibrosis, these results demonstrate how small molecule inhibitors of DUBs can exert therapeutic effects through modulation of HIF transcription factors in liver disease. Furthermore, inhibition of HIF activity using UCHL1 inhibitors may represent a therapeutic opportunity with other HIF-related pathologies.
肝纤维化是大多数慢性肝病中细胞外基质蛋白过度积累的结果。在细胞水平上,肝纤维化与肝星状细胞(HSCs)的激活有关,HSCs 转分化为具有收缩、增殖和促纤维化功能的肌成纤维细胞样表型。HSC 转分化诱导全基因组基因表达的变化,使细胞获得其促纤维化功能。我们之前已经确定,去泛素化酶泛素 C 端水解酶 1(UCHL1)在 HSC 激活后高度诱导;然而,其去泛素化活性的细胞靶标定义不明确。在这里,我们描述了 UCHL1 在调节缺氧诱导因子 1(HIF1)水平和活性中的作用,HIF1 是一种氧敏感的转录因子,在 HSC 激活和肝纤维化过程中。HIF1 在 HSC 激活期间升高,并促进促纤维化介质 HIF 靶基因的表达。HIF1α表达增加与 UCHL1 mRNA 和蛋白在 HSC 激活时的诱导相关。UCHL1 的遗传缺失或化学抑制通过降低 HIF1α 水平损害了 HIF 活性。此外,我们的机制研究表明,UCHL1 通过特异性切割降解泛素链来提高 HIF 活性,增加促纤维化基因表达水平并增加增殖速率。正如我们还表明,UCHL1 抑制在纤维化的临床前 3D 人肝切片模型中减弱纤维化,这些结果表明 DUBs 的小分子抑制剂如何通过调节肝脏疾病中的 HIF 转录因子发挥治疗作用。此外,使用 UCHL1 抑制剂抑制 HIF 活性可能代表其他与 HIF 相关的病理的治疗机会。