Universität zu Köln

Mand, Matthias, Holthaus, Michelle ORCID: 0000-0002-6207-1141, Lieder, Helmut R. ORCID: 0000-0003-2237-9050, Kleinbongard, Petra ORCID: 0000-0003-3576-3772, Conradi, Lenard, Wahlers, Thorsten ORCID: 0000-0002-8437-4119, Paunel-Görgülü, Adnana ORCID: 0000-0001-6467-3544 and Simon,, Michel (2025). Peptidylarginine deiminase 4 deficiency alleviates hypoxia/reoxygenation-induced cardiomyocyte injury. PLOS One, 20 (9). pp. 1-18. PLoS Public Library of Science. ISSN 1932-6203

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Abstract

[Artikel-Nr.: e0330864] Background: Cardiac ischemia reperfusion (I/R) injury is a serious consequence of reperfusion therapy for myocardial infarction (MI). Peptidylarginine deiminase 4 (PAD4) is a calcium-dependent enzyme that catalyzes the citrullination of proteins. In previous studies, PAD4 inhibition protected distinct organs from I/R injury by preventing the formation of neutrophil extracellular traps (NETs) and attenuating inflammatory responses. Here, we hypothesized that cardiomyocyte PAD4 expression may play a role in acute I/R injury. Methods: Infarct size was determined in isolated pressure constant-perfused hearts from WT and PAD4-deficient (PAD4 -/- ) mice. Additionally, extracellular reactive oxygen species (ROS) and cell viability were quantified in freshly isolated adult cardiomyocytes exposed to hypoxia followed by reoxygenation (H/R). Resistance to oxidative stress was proven in both genotypes by treatment of neonatal cardiomyocytes with hydrogen peroxide. Moreover, intracellular ROS formation, ATP production, mitochondrial membrane polarisation, caspase-3 activation, and cell viability were quantified after hypoxia followed by 4 h and 20 h of reoxygenation, respectively. The PAD4-specific inhibitor GSK484 was added before H/R or at reperfusion in certain experiments. Results: Infarct size was smaller in PAD4 -/- hearts following I/R when compared to the WT. Similarly, the viability of adult and neonatal PAD4 -/- cardiomyocytes was better preserved after H/R, accompanied by reduced ROS formation. PAD4 deficiency maintained mitochondrial integrity and protected neonatal cardiomyocytes against apoptosis. However, these cells did not exhibit resistance to hydrogen peroxide-induced cell death, indicating an unaltered antioxidative state. Whereas pharmacological PAD4 inhibition by GSK484 before H/R sustained intracellular ATP levels in WT cardiomyocytes, administration of GSK484 at reoxygenation did not. However, GSK484 significantly improved cardiomyocyte metabolic activity, regardless of the time of administration. Conclusions: Our study is the first to demonstrate that PAD4 expression in cardiomyocytes contributes to H/R injury independent of systemic immune responses and NETs. Consequently, PAD4 may serve as a therapeutic target to alleviate I/R injury.

Item Type:	Article
Creators:	Creators Email ORCID ORCID Put Code Mand, Matthias UNSPECIFIED UNSPECIFIED UNSPECIFIED Holthaus, Michelle UNSPECIFIED https://orcid.org/0000-0002-6207-1141 UNSPECIFIED Lieder, Helmut R. UNSPECIFIED https://orcid.org/0000-0003-2237-9050 UNSPECIFIED Kleinbongard, Petra UNSPECIFIED https://orcid.org/0000-0003-3576-3772 UNSPECIFIED Conradi, Lenard UNSPECIFIED UNSPECIFIED UNSPECIFIED Wahlers, Thorsten UNSPECIFIED https://orcid.org/0000-0002-8437-4119 UNSPECIFIED Paunel-Görgülü, Adnana UNSPECIFIED https://orcid.org/0000-0001-6467-3544 UNSPECIFIED Simon,, Michel UNSPECIFIED UNSPECIFIED UNSPECIFIED
URN:	urn:nbn:de:hbz:38-811518
Identification Number:	10.1371/journal.pone.0330864
Journal or Publication Title:	PLOS One
Volume:	20
Number:	9
Page Range:	pp. 1-18
Number of Pages:	18
Date:	10 September 2025
Publisher:	PLoS Public Library of Science
ISSN:	1932-6203
Language:	English
Faculty:	Faculty of Medicine
Divisions:	Faculty of Medicine > Chirurgie > Klinik und Poliklinik für Herzchirurgie
Subjects:	Medical sciences Medicine
['eprint_fieldname_oa_funders' not defined]:	Publikationsfonds UzK
Refereed:	Yes
URI:	http://kups.ub.uni-koeln.de/id/eprint/81151