New therapeutic approaches to target myofibroblast activation in liver pathologies: fibrosis and cholangiocarcinoma

Abstract

[eng] HYPOTHESES: Iron accumulation and senescence mediate fibrosis progression in kidney and lung. In the liver, iron overload, specifically in hepatocytes, has been linked to hepatic diseases such as hereditary hemochromatosis, MASLD or ALD. Therefore, we hypothesized that iron and senescence could have a role in liver fibrosis progression, and in this context, an iron chelator could become a potential therapeutic approach. Preliminary results from our group showed that in CCA, TGF-β has tumour suppressor effects. However, TGF-β has been linked to protumorigenic effects in the TME. Previous literature indicated that NOX4 plays a role downstream TGF-β in hepatic stellate cell activation towards matrix-producing myofibroblasts. This evidence suggested that NOX4 could be playing a role in CAF activation in CCA. Since CAF support and promote CCA progression and chemoresistance, we hypothesized that targeting NOX4 could have therapeutic effects.

OBJECTIVES: Analysis of iron chelation as a new therapeutic approach in liver fibrosis. To evaluate whether iron accumulates, correlating with characteristics of cell senescence, in a mouse model of liver fibrosis induced by CCl4. Evaluation of iron chelation by deferiprone as a therapeutic approach for liver fibrosis in vivo. Analysis of the efficacy of deferiprone to attenuate iron accumulation and senescence in vitro. To explore the translational relevance of iron accumulation and senescence in the development of liver fibrosis in patients.

Analysis of the potential therapeutic role of targeting NOX4 in hepatic stellate cell activation. Implications in cholangiocarcinoma. Analysis of NOX4 expression in CCA tumour and stromal cells. Generation and characterization of a syngeneic orthotopic CCA model in wild type and NOX4-/- mice to evaluate the role of stromal NOX4 on tumour progression. Generation of loss of function experimental models of NOX4 in human hepatic stellate cells by CRISPR-Cas9 and siRNA to analyse transdifferentiation into myofibroblasts in vitro. Evaluation of the efficacy of NOX4 inhibitors on HSC transdifferentiation in vitro. Evaluation of the efficacy of NOX4 inhibitors on tumour progression in a murine AKT-YAP hydrodynamic tail vein injection CCA model.