Methylation of the Hippo effector YAP by the methyltransferase SETD7 drives myocardial ischaemic injury: a translational study

Samuele Ambrosini; Fabrizio Montecucco; Detmar Kolijn; Daniela Pedicino; Alexander Akhmedov; Shafeeq A Mohammed; Melissa Herwig; Era Gorica; Petra L Szabó; Lukas Weber; Giulio Russo; Ramona Vinci; Christian M Matter; Giovanna Liuzzo; Peter J Brown; Fabio M V Rossi; Giovanni G Camici; Sebastiano Sciarretta; Antonio P Beltrami; Filippo Crea; Bruno Podesser; Thomas F Lüscher; Attila Kiss; Frank Ruschitzka; Nazha Hamdani; Sarah Costantino; Francesco Paneni

doi:10.1093/cvr/cvac102

Methylation of the Hippo effector YAP by the methyltransferase SETD7 drives myocardial ischaemic injury: a translational study

Cardiovasc Res. 2023 Jan 18;118(17):3374-3385. doi: 10.1093/cvr/cvac102.

Authors

Samuele Ambrosini¹, Fabrizio Montecucco^{2

3}, Detmar Kolijn^{4

5

6}, Daniela Pedicino^{7

8}, Alexander Akhmedov¹, Shafeeq A Mohammed¹, Melissa Herwig^{4

5

6}, Era Gorica^{1

9}, Petra L Szabó¹⁰, Lukas Weber¹⁰, Giulio Russo^{7

8}, Ramona Vinci^{7

8}, Christian M Matter^{1

11}, Giovanna Liuzzo^{7

8}, Peter J Brown¹², Fabio M V Rossi¹³, Giovanni G Camici^{1

11

14}, Sebastiano Sciarretta^{15

16}, Antonio P Beltrami^{17

18}, Filippo Crea^{7

8}, Bruno Podesser¹⁰, Thomas F Lüscher^{1

19}, Attila Kiss¹⁰, Frank Ruschitzka¹¹, Nazha Hamdani^{4

5

6}, Sarah Costantino^{1

11}, Francesco Paneni^{1

11

14}

Affiliations

¹ Center for Molecular Cardiology, University of Zürich, Wagistrasse 12, 8952 Schlieren, Switzerland.
² First Clinic of Internal Medicine, Department of Internal Medicine, University of Genoa, viale Benedetto XV, 16132, Genoa, Italy.
³ IRCCS Ospedale Policlinico San Martino Genova-Italian Cardiovascular Network, Largo Rosanna Benzi, 10, 16132 Genova, Italy.
⁴ Institute of Physiology, Ruhr University, Universitätsstraße 150, 44801 Bochum, Germany.
⁵ Molecular and Experimental Cardiology, Ruhr University, Universitätsstraße 150, 44801 Bochum, Germany.
⁶ Department of Cardiology, St-Josef Hospital, Ruhr University, Gudrunstraße 56, 44791 Bochum, Germany.
⁷ Dipartimento di Scienze Cardiovascolari e Toraciche, Università Cattolica del Sacro Cuore, Largo Agostino Gemelli 8, 00168, Rome, Italy.
⁸ Fondazione Policlinico Universitario A. Gemelli IRCCS, Via Giuseppe Moscati, 31, 00168 Rome, Italy.
⁹ Department of Pharmacy, University of Pisa, via Bonanno, 6, I-56126 Pisa, Italy.
¹⁰ Ludwig-Boltzmann-Institute for Cardiovascular Research at the Center for Biomedical Research, Medical University of Vienna, Währinger Gürtel 18-20A-1090 Wien, Austria.
¹¹ University Heart Center, Cardiology, University Hospital Zurich, Rämistrasse 100, 8091 Zürich, Switzerland.
¹² Structural Genomics Consortium, Univerity of Toronto, MaRS South Tower, Suite 700101 College Street, Toronto, ON M5G 1L7, Canada.
¹³ Biomedical Research Centre, University of British Columbia, Vancouver, BC V6T 1Z3, Canada.
¹⁴ Department of Research and Education, University Hospital Zurich, Rämistrasse 100, 8091 Zürich, Switzerland.
¹⁵ Dipartimento di Scienze e Biotecnologie Medico-Chirurgiche, Sapienza Università di Roma, C.so della Repubblica, 79, 04100 Latina LT, Italy.
¹⁶ Department of AngioCardioNeurology, IRCCS Neuromed, Via Atinense, 18, 86077 Pozzilli IS, Italy.
¹⁷ University of Udine, Piazzale Massimiliano Kolbe, 4, 33100 Udine, Italy.
¹⁸ Institute of Clinical Pathology, Academic Hospital "Santa Maria della Misericordia", ASUFC, 33100 Udine, Italy.
¹⁹ Royal Brompton & Harefield Hospitals, Imperial College and King's College, Sydney Street, London SW3 6NP, UK.

PMID: 35709329
DOI: 10.1093/cvr/cvac102

Abstract

Aims: Methylation of non-histone proteins is emerging as a central regulatory mechanism in health and disease. The methyltransferase SETD7 has shown to methylate and alter the function of a variety of proteins in vitro; however, its function in the heart is poorly understood. The present study investigates the role of SETD7 in myocardial ischaemic injury.

Methods and results: Experiments were performed in neonatal rat ventricular myocytes (NRVMs), SETD7 knockout mice (SETD7-/-) undergoing myocardial ischaemia/reperfusion (I/R) injury, left ventricular (LV) myocardial samples from patients with ischaemic cardiomyopathy (ICM), and peripheral blood mononuclear cells (PBMCs) from patients with ST-elevation MI (STEMI). We show that SETD7 is activated upon energy deprivation in cultured NRVMs and methylates the Hippo pathway effector YAP, leading to its cytosolic retention and impaired transcription of antioxidant genes manganese superoxide dismutase (MnSOD) and catalase (CAT). Such impairment of antioxidant defence was associated with mitochondrial reactive oxygen species (mtROS), organelle swelling, and apoptosis. Selective pharmacological inhibition of SETD7 by (R)-PFI-2 restored YAP nuclear localization, thus preventing mtROS, mitochondrial damage, and apoptosis in NRVMs. In mice, genetic deletion of SETD7 attenuated myocardial I/R injury, mtROS, and LV dysfunction by restoring YAP-dependent transcription of MnSOD and CAT. Moreover, in cardiomyocytes isolated from I/R mice and ICM patients, (R)-PFI-2 prevented mtROS accumulation, while improving Ca2+-activated tension. Finally, SETD7 was up-regulated in PBMCs from STEMI patients and negatively correlated with MnSOD and CAT.

Conclusion: We show a methylation-dependent checkpoint regulating oxidative stress during myocardial ischaemia. SETD7 inhibition may represent a valid therapeutic strategy in this setting.

Keywords: Hippo pathway; Ischaemic heart disease; Myocardial infarction; Oxidative stress; Protein methylation.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Antioxidants*
Apoptosis
Histone-Lysine N-Methyltransferase* / genetics
Histone-Lysine N-Methyltransferase* / metabolism
Humans
Leukocytes, Mononuclear / metabolism
Methylation
Mice
Mice, Knockout
Myocytes, Cardiac / metabolism
Rats
ST Elevation Myocardial Infarction* / metabolism

Substances

(R)-PFI-2
Antioxidants
Histone-Lysine N-Methyltransferase
Setd7 protein, mouse
Setd7 protein, rat
SETD7 protein, human