Targeting peroxiredoxin 2 prevents hepatocarcinogenesis in metabolic liver disease models
Emilie Crouchet, Eugénie Schaeffer, Marine A. Oudot, Julien Moehlin, Cloé Gadenne, Frank Jühling, Hussein El Saghire, Naoto Fujiwara, Shijia Zhu, Fahmida Akter Rasha, Sarah C. Durand, Anouk Charlot, Clara Ponsolles, Romain Martin, Nicolas Brignon, Fabio Del Zompo, Laura Meiss-Heydmann, Marie Parnot, Nourdine Hamdane, Danijela Heide, Jenny Hetzer, Mathias Heikenwälder, Emanuele Felli, Patrick Pessaux, Nathalie Pochet, Joffrey Zoll, Brian Cunniff, Yujin Hoshida, Laurent Mailly, Thomas F. Baumert, Catherine Schuster
Emilie Crouchet, Eugénie Schaeffer, Marine A. Oudot, Julien Moehlin, Cloé Gadenne, Frank Jühling, Hussein El Saghire, Naoto Fujiwara, Shijia Zhu, Fahmida Akter Rasha, Sarah C. Durand, Anouk Charlot, Clara Ponsolles, Romain Martin, Nicolas Brignon, Fabio Del Zompo, Laura Meiss-Heydmann, Marie Parnot, Nourdine Hamdane, Danijela Heide, Jenny Hetzer, Mathias Heikenwälder, Emanuele Felli, Patrick Pessaux, Nathalie Pochet, Joffrey Zoll, Brian Cunniff, Yujin Hoshida, Laurent Mailly, Thomas F. Baumert, Catherine Schuster
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Research Article Hepatology Oncology

Targeting peroxiredoxin 2 prevents hepatocarcinogenesis in metabolic liver disease models

Abstract

Treatment options for advanced liver disease and hepatocellular carcinoma (HCC) are limited, and strategies to prevent HCC development are lacking. Aiming to discover therapeutic targets, we combined genome-wide transcriptomic analysis of liver tissues from patients with advanced liver disease and HCC and a cell-based system predicting liver disease progression and HCC risk. Computational analysis predicted peroxiredoxin 2 (PRDX2) as a candidate gene mediating hepatocarcinogenesis and HCC risk. Analysis of tissues from patients with HCC confirmed a perturbed expression of PRDX2 in cancer. In vivo perturbation studies in mouse models for hepatocarcinogenesis driven by metabolic dysfunction–associated steatohepatitis showed that specific Prdx2 KO in hepatocytes improved metabolic liver functions, restored AMPK activity, and prevented HCC development by suppressing oncogenic signaling. Perturbation studies in HCC cell lines, a cell line–derived xenograft mouse model, and patient-derived HCC spheroids revealed that PRDX2 also mediates cancer initiation, cancer cell proliferation, and survival through its antioxidant activity. Targeting PRDX2 may therefore be a strategy to prevent HCC development in metabolic liver disease.

Authors

Emilie Crouchet, Eugénie Schaeffer, Marine A. Oudot, Julien Moehlin, Cloé Gadenne, Frank Jühling, Hussein El Saghire, Naoto Fujiwara, Shijia Zhu, Fahmida Akter Rasha, Sarah C. Durand, Anouk Charlot, Clara Ponsolles, Romain Martin, Nicolas Brignon, Fabio Del Zompo, Laura Meiss-Heydmann, Marie Parnot, Nourdine Hamdane, Danijela Heide, Jenny Hetzer, Mathias Heikenwälder, Emanuele Felli, Patrick Pessaux, Nathalie Pochet, Joffrey Zoll, Brian Cunniff, Yujin Hoshida, Laurent Mailly, Thomas F. Baumert, Catherine Schuster

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Figure 8

Targeting PRDX2 in cancer cells increases oxidative stress, reduces cell proliferation, and sensitizes cancer cells to apoptosis.

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Targeting PRDX2 in cancer cells increases oxidative stress, reduces cell...
(A) PRDX2 KD increases ROS production in Huh7 cells. ROS generation was measured in Huh7 cells upon oxidative stress induced by H202 (300 μM, 4 hours), by flow cytometry. One representative histogram is shown. The graph shows mean ± SD of the percentage of delta mean of fluorescence from 3 independent experiments performed in triplicate (n = 9). ***P < 0.001; ****P < 0.0001 (ordinary 1-way ANOVA followed by Tukey’s multiple-comparison test). (B) PRDX2 KD reduces cell proliferation. Huh7 cell proliferation and cell cycle profile were assessed by flow cytometry and costaining with Edu (cell proliferation) and Fx cell cycle (total DNA). One representative example of cell cycle profile is shown (blue = G0/G1; yellow = S, and green = G2/M). The graphs show mean ± SD of the percentage of proliferative cells (Edu+ cells) and of the percentage of cells in the different cell cycle steps of 3 independent experiments performed in 4 replicates (n = 12). ****P < 0.0001 (Mann-Whitney U test). (C) PRDX2 KD impairs mitochondrial function. Mitochondrial respiration in Huh7 cells was assessed in a 2-chamber respirometer Oroboros Oxygraph-2k at 37°C. The graph shows time course of oxygen consumption upon successive activation/inhibition of the different mitochondria complexes. One representative experiment out of 3 is shown. (D) PRDX2 KD increases ER stress. ER stress was assessed by Western blot analysis of the GRP78 marker. The graph shows mean ± SD of normalized protein intensity (normalization to total proteins) of 2 independent experiments performed in duplicates (n = 4). Exact P value is indicated (Mann-Whitney U test). (E and F) Targeting PRDX2 sensitizes cancer cells to apoptosis upon oxidative stress (H2O2 300 μM, 6 hours). Apoptosis was assessed by (E) Western blot analysis of caspase-3 and cleaved caspase-3 (graph shows mean ± SD of normalized protein intensity — normalization to total proteins — of 3 independent experiments, n = 3) and (F) by detecting activated caspase-3/7 using CellEvent caspase-3/7 detection reagent. Activated caspase is shown in green. Nuclei were counterstained in blue (DAPI). Scale bar: 200 μm. One representative experiment out of 3 is shown.