Cholesterol crystals use complement to increase NLRP3 signaling pathways in coronary and carotid atherosclerosis
Nathalie Niyonzima,
Siril S. Bakke,
Ida Gregersen,
Sverre Holm,
Øystein Sandanger,
Hilde L. Orrem,
Bjørnar Sporsheim,
Liv Ryan,
Xiang Yi Kong,
Tuva Børresdatter Dahl,
Mona Skjelland,
Kirsten Krohg Sørensen,
Anne Mari Rokstad,
Arne Yndestad,
Eicke Latz,
Lars Gullestad,
Geir Ø. Andersen,
Jan Kristian Damås,
Pål Aukrust,
Tom E. Mollnes,
Bente Halvorsen,
Terje Espevik
Affiliations
Nathalie Niyonzima
Centre of Molecular Inflammation Research, and Department of Clinical and Molecular Medicine, Norwegian University of Science and Technology, Olav Kyrres gate 17, Trondheim 7030, Norway
Siril S. Bakke
Centre of Molecular Inflammation Research, and Department of Clinical and Molecular Medicine, Norwegian University of Science and Technology, Olav Kyrres gate 17, Trondheim 7030, Norway
Ida Gregersen
Research Institute of Internal Medicine, Oslo University Hospital, Norway; Institute of Clinical Medicine, Faculty of Medicine, University of Oslo, Norway
Sverre Holm
Research Institute of Internal Medicine, Oslo University Hospital, Norway
Øystein Sandanger
Research Institute of Internal Medicine, Oslo University Hospital, Norway; Institute of Clinical Medicine, Faculty of Medicine, University of Oslo, Norway
Hilde L. Orrem
Division of Emergencies and Critical Care, Oslo University Hospital, Oslo, Norway
Bjørnar Sporsheim
Centre of Molecular Inflammation Research, and Department of Clinical and Molecular Medicine, Norwegian University of Science and Technology, Olav Kyrres gate 17, Trondheim 7030, Norway
Liv Ryan
Centre of Molecular Inflammation Research, and Department of Clinical and Molecular Medicine, Norwegian University of Science and Technology, Olav Kyrres gate 17, Trondheim 7030, Norway
Xiang Yi Kong
Research Institute of Internal Medicine, Oslo University Hospital, Norway
Tuva Børresdatter Dahl
Research Institute of Internal Medicine, Oslo University Hospital, Norway
Mona Skjelland
Institute of Clinical Medicine, Faculty of Medicine, University of Oslo, Norway; Department of Neurology, Oslo University Hospital, Norway
Kirsten Krohg Sørensen
Research Institute of Internal Medicine, Oslo University Hospital, Norway; Institute of Clinical Medicine, Faculty of Medicine, University of Oslo, Norway
Anne Mari Rokstad
Centre of Molecular Inflammation Research, and Department of Clinical and Molecular Medicine, Norwegian University of Science and Technology, Olav Kyrres gate 17, Trondheim 7030, Norway
Arne Yndestad
Research Institute of Internal Medicine, Oslo University Hospital, Norway; Institute of Clinical Medicine, Faculty of Medicine, University of Oslo, Norway
Eicke Latz
Centre of Molecular Inflammation Research, and Department of Clinical and Molecular Medicine, Norwegian University of Science and Technology, Olav Kyrres gate 17, Trondheim 7030, Norway; Institute of Innate Immunity, Biomedical Center, University of Bonn, Germany
Lars Gullestad
Department of Cardiology, Oslo University Hospital, Norway; KG Jebsen Center for Cardiac Research, and Center for Heart Failure Research, Oslo University Hospital, Norway
Geir Ø. Andersen
Department of Cardiology, Oslo University Hospital, Norway
Jan Kristian Damås
Centre of Molecular Inflammation Research, and Department of Clinical and Molecular Medicine, Norwegian University of Science and Technology, Olav Kyrres gate 17, Trondheim 7030, Norway
Pål Aukrust
Research Institute of Internal Medicine, Oslo University Hospital, Norway; Institute of Clinical Medicine, Faculty of Medicine, University of Oslo, Norway; Section of Clinical Immunology and Infectious Diseases, Oslo University Hospital, Norway
Tom E. Mollnes
Centre of Molecular Inflammation Research, and Department of Clinical and Molecular Medicine, Norwegian University of Science and Technology, Olav Kyrres gate 17, Trondheim 7030, Norway; Department of Immunology, Oslo University Hospital, Norway; KG Jebsen TREC, Department of Clinical Medicine, University of Tromsø, Norway; Research Laboratory, Norland Hospital, Norway
Bente Halvorsen
Research Institute of Internal Medicine, Oslo University Hospital, Norway; Institute of Clinical Medicine, Faculty of Medicine, University of Oslo, Norway
Terje Espevik
Centre of Molecular Inflammation Research, and Department of Clinical and Molecular Medicine, Norwegian University of Science and Technology, Olav Kyrres gate 17, Trondheim 7030, Norway; The Central Norway Regional Health Authority, St. Olavs Hospital HF, Norway; Corresponding author at: Centre of Molecular Inflammation Research, and Department of Clinical and Molecular Medicine, Norwegian University of Science and Technology, Olav Kyrres gate 17, Trondheim 7030, Norway.
Background: During atherogenesis, cholesterol precipitates into cholesterol crystals (CC) in the vessel wall, which trigger plaque inflammation by activating the NACHT, LRR and PYD domains-containing protein 3 (NLRP3) inflammasome. We investigated the relationship between CC, complement and NLRP3 in patients with cardiovascular disease. Methods: We analysed plasma, peripheral blood mononuclear cells (PBMC) and carotid plaques from patients with advanced atherosclerosis applying ELISAs, multiplex cytokine assay, qPCR, immunohistochemistry, and gene profiling. Findings: Transcripts of interleukin (IL)-1beta(β) and NLRP3 were increased and correlated in PBMC from patients with acute coronary syndrome (ACS). Priming of these cells with complement factor 5a (C5a) and tumour necrosis factor (TNF) before incubation with CC resulted in increased IL-1β protein when compared to healthy controls. As opposed to healthy controls, systemic complement was significantly increased in patients with stable angina pectoris or ACS. In carotid plaques, complement C1q and C5b-9 complex accumulated around CC-clefts, and complement receptors C5aR1, C5aR2 and C3aR1 were higher in carotid plaques compared to control arteries. Priming human carotid plaques with C5a followed by CC incubation resulted in pronounced release of IL-1β, IL-18 and IL-1α. Additionally, mRNA profiling demonstrated that C5a and TNF priming followed by CC incubation upregulated plaque expression of NLRP3 inflammasome components. Interpretation: We demonstrate that CC are important local- and systemic complement activators, and we reveal that the interaction between CC and complement could exert its effect by activating the NLRP3 inflammasome, thus promoting the progression of atherosclerosis.
