Characteristic ERK1/2 signaling dynamics distinguishes necroptosis from apoptosis

François Sipieter; Benjamin Cappe; Aymeric Leray; Elke De Schutter; Jolien Bridelance; Paco Hulpiau; Guy Van Camp; Wim Declercq; Laurent Héliot; Pierre Vincent; Peter Vandenabeele; Franck B Riquet

doi:10.1016/j.isci.2021.103074

Characteristic ERK1/2 signaling dynamics distinguishes necroptosis from apoptosis

iScience. 2021 Sep 2;24(9):103074. doi: 10.1016/j.isci.2021.103074. eCollection 2021 Sep 24.

Authors

François Sipieter^{1

2

3}, Benjamin Cappe^{1

2}, Aymeric Leray⁴, Elke De Schutter^{1

2

5}, Jolien Bridelance^{1

2}, Paco Hulpiau⁶, Guy Van Camp^{5

7}, Wim Declercq^{1

2}, Laurent Héliot⁸, Pierre Vincent⁹, Peter Vandenabeele^{1

2}, Franck B Riquet^{1

2

3}

Affiliations

¹ Molecular Signaling and Cell Death Unit, Department of Biomedical Molecular Biology, Ghent University, Ghent, Belgium.
² Molecular Signaling and Cell Death Unit, VIB Center for Inflammation Research, Technologiepark 71, Zwijnaarde, 9052 Ghent, Belgium.
³ Université de Lille, Lille, France.
⁴ Laboratoire Interdisciplinaire Carnot De Bourgogne, UMR 6303 CNRS-Université de Bourgogne Franche-Comté, 21078 Dijon, France.
⁵ Center of Medical Genetics, University of Antwerp and Antwerp University Hospital, Prins Boudewijnlaan 43/6, Edegem, 2650 Antwerp, Belgium.
⁶ Data Mining and Modeling for Biomedicine (DaMBi), VIB Center for Inflammation Research, Ghent, Belgium.
⁷ Center for Oncological Research, University of Antwerp and Antwerp University Hospital, Universiteitsplein 1, Wilrijk, 2610 Antwerp, Belgium.
⁸ Team Biophotonique Cellulaire Fonctionnelle, Laboratoire de Physique des Lasers, Atomes et Molécules (PhLAM), CNRS UMR 8523, 59655 Villeneuve d'Ascq, France.
⁹ Sorbonne Université, CNRS, Neurobiology of Adaptative Processes, UMR8256, 75005 Paris, France.

Abstract

ERK1/2 involvement in cell death remains unclear, although many studies have demonstrated the importance of ERK1/2 dynamics in determining cellular responses. To untangle how ERK1/2 contributes to two cell death programs, we investigated ERK1/2 signaling dynamics during hFasL-induced apoptosis and TNF-induced necroptosis in L929 cells. We observed that ERK1/2 inhibition sensitizes cells to apoptosis while delaying necroptosis. By monitoring ERK1/2 activity by live-cell imaging using an improved ERK1/2 biosensor (EKAR4.0), we reported differential ERK1/2 signaling dynamics between cell survival, apoptosis, and necroptosis. We also decrypted a temporally shifted amplitude- and frequency-modulated (AM/FM) ERK1/2 activity profile in necroptosis versus apoptosis. ERK1/2 inhibition, which disrupted ERK1/2 signaling dynamics, prevented TNF and IL-6 gene expression increase during TNF-induced necroptosis. Using an inducible cell line for activated MLKL, the final executioner of necroptosis, we showed ERK1/2 and its distinctive necroptotic ERK1/2 activity dynamics to be positioned downstream of MLKL.

Keywords: Biological sciences; Biomolecular engineering; Cell biology.