PI: Silvia Soddu
Other IRE Principal Collaborators Involved: Piaggio Giulia

Project duration: 24 months

Project code: PNRR-MAD-2022-12375670

Non-alcoholic fatty liver disease has reached global epidemic proportion becoming the most cause of chronic liver diseases (CLDs). CLD progression toward cirrhosis and hepatocellular carcinoma is linked to necroinflammation and liver fibrosis. However, the molecular determinants of these events are still unclear and candidate biomarkers for prevention and treatment are under intense investigation.

We hypothesize that a successful identification of candidate biomarkers for liver fibrosis cannot disregard the evaluation of the entire hepatic microenvironment. Traditional preclinical platforms, including 3D organoid cell cultures and their xenografts, take into account specific cell populations but do not consider the real complexity of the tissue environment. Instead, studies comparing liver biopsies are limited by the long and non-homogenous evolution of the disease. Here, we propose to overcome these limitations by performing omics-analyses on organotypic slice cultures obtained by cutting fresh human livers from patients that undergo liver surgery for metastatic disease or other pathologies. These human liver slice cultures will be treated in vitro with anti- and pro-fibrotic agents and analyzed in the following week by time-lapse, multiple immune fluorescence for known fibrosis pathways, single-cell-sequencing or bulk RNAseq. This type of analyses will be combined with a hypothesis-driven strategy based on the generation and study of a liver specific HIPK2 KO mouse model. We expect to identify actionable targets and strategies for the prevention and treatment of liver fibrosis.

This project will integrate the knowledge of basic and translational scientists (UO1), clinicians (U01, UO3, and UO4), and expert in omics techniques (UO2) and bioinformatics (UO1 and UO2).

Establishment of human liver slice cultures as a tool to study hepatic microenvironment determinants of liver fibrosis.

Generation of a liver-specific HIPK2 knockout mouse model to establish the contribution of HIPK2 in liver fibrosis

Pilot validation of HIPK2 and new emerging biomarkers of liver fibrosis

Overall, this proposal has the likelihood of identifying novel biomarker(s) for liver fibrosis prevention and treatment. This will pave the way of next clinical evaluation to validate the use of the identified biomarker(s) in different CDLs, to predict progression into cirrhosis, hepatocellular cancer, and/or induction of metastasis-permissive liver. In addition, it opens the possibility of evaluating whether the biomarker(s) might be targets for anti-fibrotic therapies.