Hepatocellular carcinoma chemoprevention by targeting the angiotensin-converting enzyme and EGFR transactivation
Emilie Crouchet, Shen Li, Mozhdeh Sojoodi, Simonetta Bandiera, Naoto Fujiwara, Hussein El Saghire, Shijia Zhu, Tongqi Qian, Fahmida Akter Rasha, Fabio Del Zompo, Stephen C. Barrett, Eugénie Schaeffer, Marine A. Oudot, Clara Ponsolles, Sarah C. Durand, Sarani Ghoshal, Gunisha Arora, Fabio Giannone, Raymond T. Chung, Nevena Slovic, Nicolaas Van Renne, Emanuele Felli, Patrick Pessaux, Joachim Lupberger, Nathalie Pochet, Catherine Schuster, Kenneth K. Tanabe, Yujin Hoshida, Bryan C. Fuchs, Thomas F. Baumert
Emilie Crouchet, Shen Li, Mozhdeh Sojoodi, Simonetta Bandiera, Naoto Fujiwara, Hussein El Saghire, Shijia Zhu, Tongqi Qian, Fahmida Akter Rasha, Fabio Del Zompo, Stephen C. Barrett, Eugénie Schaeffer, Marine A. Oudot, Clara Ponsolles, Sarah C. Durand, Sarani Ghoshal, Gunisha Arora, Fabio Giannone, Raymond T. Chung, Nevena Slovic, Nicolaas Van Renne, Emanuele Felli, Patrick Pessaux, Joachim Lupberger, Nathalie Pochet, Catherine Schuster, Kenneth K. Tanabe, Yujin Hoshida, Bryan C. Fuchs, Thomas F. Baumert
View: Text | PDF
Research Article Hepatology

Hepatocellular carcinoma chemoprevention by targeting the angiotensin-converting enzyme and EGFR transactivation

Abstract

Hepatocellular carcinoma (HCC) is a leading cause of death among cirrhotic patients, for which chemopreventive strategies are lacking. Recently, we developed a simple human cell-based system modeling a clinical prognostic liver signature (PLS) predicting liver disease progression and HCC risk. In a previous study, we applied our cell-based system for drug discovery and identified captopril, an approved angiotensin converting enzyme (ACE) inhibitor, as a candidate compound for HCC chemoprevention. Here, we explored ACE as a therapeutic target for HCC chemoprevention. Captopril reduced liver fibrosis and effectively prevented liver disease progression toward HCC development in a diethylnitrosamine (DEN) rat cirrhosis model and a diet-based rat model for nonalcoholic steatohepatitis–induced (NASH-induced) hepatocarcinogenesis. RNA-Seq analysis of cirrhotic rat liver tissues uncovered that captopril suppressed the expression of pathways mediating fibrogenesis, inflammation, and carcinogenesis, including epidermal growth factor receptor (EGFR) signaling. Mechanistic data in liver disease models uncovered a cross-activation of the EGFR pathway by angiotensin. Corroborating the clinical translatability of the approach, captopril significantly reversed the HCC high-risk status of the PLS in liver tissues of patients with advanced fibrosis. Captopril effectively prevents fibrotic liver disease progression toward HCC development in preclinical models and is a generic and safe candidate drug for HCC chemoprevention.

Authors

Emilie Crouchet, Shen Li, Mozhdeh Sojoodi, Simonetta Bandiera, Naoto Fujiwara, Hussein El Saghire, Shijia Zhu, Tongqi Qian, Fahmida Akter Rasha, Fabio Del Zompo, Stephen C. Barrett, Eugénie Schaeffer, Marine A. Oudot, Clara Ponsolles, Sarah C. Durand, Sarani Ghoshal, Gunisha Arora, Fabio Giannone, Raymond T. Chung, Nevena Slovic, Nicolaas Van Renne, Emanuele Felli, Patrick Pessaux, Joachim Lupberger, Nathalie Pochet, Catherine Schuster, Kenneth K. Tanabe, Yujin Hoshida, Bryan C. Fuchs, Thomas F. Baumert

×

Figure 5

Captopril prevents HCC development by targeting the angiotensin-induced EGFR transactivation.

Options: View larger image (or click on image) Download as PowerPoint
Captopril prevents HCC development by targeting the angiotensin-induced ...
(A) Effect of captopril on receptor tyrosine kinase (RTK) phosphorylation in the cell-based model. Heatmap shows the significance of induction (yellow) or suppression (purple) of protein phosphorylation in drug-treated samples compared with untreated controls. Results show means from 3 independent experiments performed in duplicate. (B) Captopril treatment decreases EGFR phosphorylation in HCV-infected cells. Panphosphorylation of EGFR was assessed using phosphoarray. Results are shown as mean ± SEM of integrated dot blot densities from 3 independent experiments performed in duplicate (n = 6). ***P < 0.001, unpaired t test. (C) EGFR is a downstream effector of Ang II. Panphosphorylation of EGFR was assessed using phosphoarray in Huh7.5.1dif cells. The graph shows the quantification of dot blot intensities (fold change of phosphorylated EGFR). One representative experiment out of 2 is shown. (D and E) Captopril treatment significantly repressed EGFR and downstream MAPK pathway genes induced in response to HCV infection at the single-cell level. Heatmaps show core EGFR signaling genes that are modulated by captopril treatment (blue) or HCV infection (red), both (green) or none (yellow), as defined by “leading-edge” genes driving the enrichment score in GSEAs. The effect of captopril on these genes is indicated by Z scores (middle row), while the effect of HCV infection is indicated by the Pearson correlation of the expression levels with the HCV viral load (top row). Significance of gene expression modulation was determined using the hypergeometric test. (F and G) EGFR activation is suppressed by captopril treatment in a rat model for fibrosis and HCC. (F) Captopril reverses 2 EGF target gene signature in vivo (GSEA). (G). Western blot analysis showing Erk1/2, JNK, and p38 expression and phosphorylation in the liver of animals (3 animals per group). See complete unedited blots in the supplemental material.