Niemann-Pick C1 protein regulates platelet membrane-associated calcium ion signaling in thrombo-occlusive diseases in mice

Mailin-Christin Manke; Anna Roslan; Britta Walker; Patrick Münzer; Ferdinand Kollotzek; Bing Peng; Stine Mencl; Cristina Coman; Rebecca D Szepanowski; Harald Schulze; Andrew P Lieberman; Florian Lang; Meinrad Gawaz; Christoph Kleinschnitz; Robert Lukowski; Robert Ahrends; Régis Bobe; Oliver Borst

doi:10.1016/j.jtha.2023.03.038

Niemann-Pick C1 protein regulates platelet membrane-associated calcium ion signaling in thrombo-occlusive diseases in mice

J Thromb Haemost. 2023 Jul;21(7):1957-1966. doi: 10.1016/j.jtha.2023.03.038. Epub 2023 Apr 11.

Authors

Mailin-Christin Manke¹, Anna Roslan², Britta Walker³, Patrick Münzer¹, Ferdinand Kollotzek¹, Bing Peng⁴, Stine Mencl⁵, Cristina Coman⁶, Rebecca D Szepanowski⁵, Harald Schulze⁷, Andrew P Lieberman⁸, Florian Lang⁹, Meinrad Gawaz³, Christoph Kleinschnitz⁵, Robert Lukowski², Robert Ahrends¹⁰, Régis Bobe¹¹, Oliver Borst¹²

Affiliations

¹ DFG Heisenberg Group Thrombocardiology; Department of Cardiology, Angiology and Cardiovascular Medicine, University of Tübingen, Germany.
² Department of Pharmacology, Toxicology and Clinical Pharmacy, University of Tübingen, Germany.
³ DFG Heisenberg Group Thrombocardiology.
⁴ Leibniz-Institut für Analytische Wissenschaften-ISAS, Dortmund, Germany; Division of Rheumatology, Department of Medicine Solna, Karolinska Institute, Karolinska University Hospital, Stockholm, Sweden.
⁵ Department of Neurology and Center for Translational Neuro- and Behavioral Sciences (C-TNBS), University Hospital Essen, University of Duisburg-Essen, Essen, Germany.
⁶ Department of Analytical Chemistry, University of Vienna, Austria.
⁷ Institute of Experimental Biomedicine, University Hospital Würzburg, Germany.
⁸ Department of Pathology, University of Michigan Medical School, USA.
⁹ Department of Physiology, University of Tübingen, Germany.
¹⁰ Leibniz-Institut für Analytische Wissenschaften-ISAS, Dortmund, Germany; Department of Analytical Chemistry, University of Vienna, Austria.
¹¹ HITh, UMR_S1176, INSERM, Université Paris-Saclay, France.
¹² DFG Heisenberg Group Thrombocardiology; Department of Cardiology, Angiology and Cardiovascular Medicine, University of Tübingen, Germany. Electronic address: oliver.borst@med.uni-tuebingen.de.

PMID: 37054918
DOI: 10.1016/j.jtha.2023.03.038

Abstract

Background: Pathophysiologic platelet activation leads to thrombo-occlusive diseases such as myocardial infarction or ischemic stroke. Niemann-Pick C1 protein (NPC1) is involved in the regulation of lysosomal lipid trafficking and calcium ion (Ca²⁺) signaling, and its genetic mutation causes a lysosomal storage disorder. Lipids and Ca²⁺ are key players in the complex orchestration of platelet activation.

Objectives: The present study aimed to determine the impact of NPC1 on Ca²⁺ mobilization during platelet activation in thrombo-occlusive diseases.

Methods: Using MK/platelet-specific knockout mice of Npc1 (Npc1^{Pf4∆/Pf4∆}), ex vivo and in vitro approaches as well as in vivo models of thrombosis, we investigated the effect of Npc1 on platelet function and thrombus formation.

Results: We showed that Npc1^{Pf4∆/Pf4∆} platelets display increased sphingosine levels and a locally impaired membrane-associated and SERCA3-dependent Ca²⁺ mobilisation compared to platelets from wildtype littermates (Npc1^lox/lox). Further, we observed decreased platelet.

Conclusion: Our findings highlight that NPC1 regulates membrane-associated and SERCA3-dependent Ca²⁺ mobilization during platelet activation and that MK/platelet-specific ablation of Npc1 protects against experimental models of arterial thrombosis and myocardial or cerebral ischemia/reperfusion injury.

Keywords: Niemann-Pick C1 protein; arterial thrombosis; calcium signaling; platelet activation; sphingolipids.

Publication types

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

MeSH terms

Animals
Calcium / metabolism
Intracellular Signaling Peptides and Proteins / genetics
Intracellular Signaling Peptides and Proteins / metabolism
Mice
Mice, Knockout
Niemann-Pick C1 Protein*
Niemann-Pick Disease, Type C* / genetics
Niemann-Pick Disease, Type C* / metabolism

Substances

Niemann-Pick C1 Protein
Calcium
Intracellular Signaling Peptides and Proteins