Hepatocellular carcinoma (HCC) is the most common primary liver cancer and a dominant cause of cancer-related death, underscoring the clinical, social and economic burden. Over the past years, a plethora of preclinical and clinical studies have been executed in the search for effective HCC therapy but to date, available treatment options are still inadequate emphasizing the need for new therapeutic options. The establishment of novel preclinical models that interrogate the mechanisms driving HCC pathogenesis and allow therapy evaluation in the context of an effective tumour microenvironment in analogue with human disease, is essential translational research to address this high medical need. HCC usually develops in a background of chronic liver injury, characterized by hypoxia, oxidative stress and necro-inflammation. NADPH oxidases (NOXs) catalyse the production of reactive oxygen species and the NOX1 isoform is upregulated and correlates with poor prognosis in human HCC. We and others have recently shown that NOX1 inhibition alters the tumour microenvironment and attenuates tumour development in a chemically induced HCC mouse model. In addition, NOX1 inhibition enhanced the activity of anti-PD1 antibody immunotherapy in experimental colorectal cancer emphasizing on the potential of NOX1 inhibitors as combination strategy in solid tumours. Here, we wish to (1) validate the therapeutic potential of NOX1 inhibition in a relevant orthotopic syngeneic HCC mouse model that allows non-invasive therapy evaluation and (2) optimize and use a humanised orthotopic HCC mouse model, including mice with a humanised immune system, and human HCC precision-cut liver slices, two highly relevant preclinical human-based models, to translate our results of NOX1 inhibition as therapy for HCC to the clinic. Our ultimate goal is to provide evidence for a clinical trial with NOX1 inhibitors, in mono- or combination therapy, in HCC patients