Martina Gast1, Bernhard H Rauch2,3, Arash Haghikia1,4, Shinichi Nakagawa4,5, Jan Haas6,7, Andrea Stroux8, David Schmidt1, Paul Schumann1, Stefan Weiss9, Lars Jensen9, Adelheid Kratzer1, Nicolle Kraenkel1, Christian Müller10,11, Daniela Börnigen10,11, Tetsuro Hirose12, Stefan Blankenberg10,11, Felicitas Escher13,14,15, Anja A Kühl16, Andreas W Kuss9, Benjamin Meder6,7,17, Ulf Landmesser1,13,18, Tanja Zeller10,11, Wolfgang Poller1,13,19. 1. Department of Cardiology, Charité-Universitätsmedizin Berlin, Campus Benjamin Franklin, Charite Centrum 11, Hindenburgdamm 30, Berlin, Germany. 2. Institute for Pharmacology, Universitätsmedizin Greifswald, Felix-Hausdorff-Strasse 3, Greifswald, Germany. 3. German Center for Cardiovascular Research (DZHK), Site Greifswald, Felix-Hausdorff-Strasse 3, Greifswald. 4. RNA Biology Laboratory, RIKEN Advanced Research Institute, Wako, Saitama, Japan. 5. Faculty of Pharmaceutical Sciences, Hokkaido University, Kita-12 jo, Nishi 6-chome, Kita-ku, Sapporo, Japan. 6. Department of Cardiology, Institute for Cardiomyopathies, University Hospital Heidelberg, Im Neuenheimer Feld 669, Heidelberg, Germany. 7. German Center for Cardiovascular Research (DZHK), Site Heidelberg, Im Neuenheimer Feld 669, Heidelberg, Germany. 8. Institute for Biometry and Clinical Epidemiology, Hindenburgdamm 30, Berlin, Germany. 9. Interfaculty Institute for Genetics and Functional Genome Research, University of Greifswald, Felix-Hausdorff-Strasse 8, Greifswald, Germany. 10. Clinic for General and Interventional Cardiology, University Heart Center Hamburg, Martinistrasse 52, Hamburg, Germany. 11. German Center for Cardiovascular Research (DZHK), Site Hamburg/Lübeck/Kiel, Martinistrasse 52, Hamburg, Germany. 12. Institute for Genetic Medicine, Hokkaido University, Sapporo, Japan. 13. German Center for Cardiovascular Research (DZHK), Site Berlin, Hindenburgdamm 30, Berlin, Germany. 14. Institute of Cardiac Diagnostics and Therapy (IKDT), Hindenburgdamm 30, Berlin, Germany. 15. Department of Cardiology CVK, Hindenburgdamm 30, Berlin, Germany. 16. iPATH.Berlin-Core Unit Immunopathology, Charité-Universitätsmedizin Berlin, Berlin, Germany. 17. Department of Genetics, Genome Technology Center, Stanford University Medical School, Stanford, CA, USA. 18. Berlin Institute of Health (BIH), Anna-Louisa-Karsch-Strasse 2, Berlin, Germany. 19. Berlin-Brandenburg Center for Regenerative Therapies (BCRT), Hindenburgdamm 30, Berlin, Germany.
Abstract
AIMS: Inflammation is a key driver of atherosclerosis and myocardial infarction (MI), and beyond proteins and microRNAs (miRs), long noncoding RNAs (lncRNAs) have been implicated in inflammation control. To obtain further information on the possible role of lncRNAs in the context of atherosclerosis, we obtained comprehensive transcriptome maps of circulating immune cells (peripheral blood mononuclear cells, PBMCs) of early onset MI patients. One lncRNA significantly suppressed in post-MI patients was further investigated in a murine knockout model. METHODS AND RESULTS: Individual RNA-sequencing (RNA-seq) was conducted on PBMCs from 28 post-MI patients with a history of MI at age ≤50 years and stable disease ≥3 months before study participation, and from 31 healthy individuals without manifest cardiovascular disease or family history of MI as controls. RNA-seq revealed deregulated protein-coding transcripts and lncRNAs in post-MI PBMCs, among which nuclear enriched abundant transcript (NEAT1) was the most highly expressed lncRNA, and the only one significantly suppressed in patients. Multivariate statistical analysis of validation cohorts of 106 post-MI patients and 85 controls indicated that the PBMC NEAT1 levels were influenced (P = 0.001) by post-MI status independent of statin intake, left ventricular ejection fraction, low-density lipoprotein or high-density lipoprotein cholesterol, or age. We investigated NEAT1-/- mice as a model of NEAT1 deficiency to evaluate if NEAT1 depletion may directly and causally alter immune regulation. RNA-seq of NEAT1-/- splenocytes identified disturbed expression and regulation of chemokines/receptors, innate immunity genes, tumour necrosis factor (TNF) and caspases, and increased production of reactive oxygen species (ROS) under baseline conditions. NEAT1-/- spleen displayed anomalous Treg and TH cell differentiation. NEAT1-/- bone marrow-derived macrophages (BMDMs) displayed altered transcriptomes with disturbed chemokine/chemokine receptor expression, increased baseline phagocytosis (P < 0.0001), and attenuated proliferation (P = 0.0013). NEAT1-/- BMDMs responded to LPS with increased (P < 0.0001) ROS production and disturbed phagocytic activity (P = 0.0318). Monocyte-macrophage differentiation was deregulated in NEAT1-/- bone marrow and blood. NEAT1-/- mice displayed aortic wall CD68+ cell infiltration, and there was evidence of myocardial inflammation which could lead to severe and potentially life-threatening structural damage in some of these animals. CONCLUSION: The study indicates distinctive alterations of lncRNA expression in post-MI patient PBMCs. Regarding the monocyte-enriched NEAT1 suppressed in post-MI patients, the data from NEAT1-/- mice identify NEAT1 as a novel lncRNA-type immunoregulator affecting monocyte-macrophage functions and T cell differentiation. NEAT1 is part of a molecular circuit also involving several chemokines and interleukins persistently deregulated post-MI. Individual profiling of this circuit may contribute to identify high-risk patients likely to benefit from immunomodulatory therapies. It also appears reasonable to look for new therapeutic targets within this circuit. Published on behalf of the European Society of Cardiology. All rights reserved.
AIMS: Inflammation is a key driver of atherosclerosis and myocardial infarction (MI), and beyond proteins and microRNAs (miRs), long noncoding RNAs (lncRNAs) have been implicated in inflammation control. To obtain further information on the possible role of lncRNAs in the context of atherosclerosis, we obtained comprehensive transcriptome maps of circulating immune cells (peripheral blood mononuclear cells, PBMCs) of early onset MI patients. One lncRNA significantly suppressed in post-MI patients was further investigated in a murine knockout model. METHODS AND RESULTS: Individual RNA-sequencing (RNA-seq) was conducted on PBMCs from 28 post-MI patients with a history of MI at age ≤50 years and stable disease ≥3 months before study participation, and from 31 healthy individuals without manifest cardiovascular disease or family history of MI as controls. RNA-seq revealed deregulated protein-coding transcripts and lncRNAs in post-MI PBMCs, among which nuclear enriched abundant transcript (NEAT1) was the most highly expressed lncRNA, and the only one significantly suppressed in patients. Multivariate statistical analysis of validation cohorts of 106 post-MI patients and 85 controls indicated that the PBMC NEAT1 levels were influenced (P = 0.001) by post-MI status independent of statin intake, left ventricular ejection fraction, low-density lipoprotein or high-density lipoprotein cholesterol, or age. We investigated NEAT1-/- mice as a model of NEAT1 deficiency to evaluate if NEAT1 depletion may directly and causally alter immune regulation. RNA-seq of NEAT1-/- splenocytes identified disturbed expression and regulation of chemokines/receptors, innate immunity genes, tumour necrosis factor (TNF) and caspases, and increased production of reactive oxygen species (ROS) under baseline conditions. NEAT1-/- spleen displayed anomalous Treg and TH cell differentiation. NEAT1-/- bone marrow-derived macrophages (BMDMs) displayed altered transcriptomes with disturbed chemokine/chemokine receptor expression, increased baseline phagocytosis (P < 0.0001), and attenuated proliferation (P = 0.0013). NEAT1-/- BMDMs responded to LPS with increased (P < 0.0001) ROS production and disturbed phagocytic activity (P = 0.0318). Monocyte-macrophage differentiation was deregulated in NEAT1-/- bone marrow and blood. NEAT1-/- mice displayed aortic wall CD68+ cell infiltration, and there was evidence of myocardial inflammation which could lead to severe and potentially life-threatening structural damage in some of these animals. CONCLUSION: The study indicates distinctive alterations of lncRNA expression in post-MI patient PBMCs. Regarding the monocyte-enriched NEAT1 suppressed in post-MI patients, the data from NEAT1-/- mice identify NEAT1 as a novel lncRNA-type immunoregulator affecting monocyte-macrophage functions and T cell differentiation. NEAT1 is part of a molecular circuit also involving several chemokines and interleukins persistently deregulated post-MI. Individual profiling of this circuit may contribute to identify high-risk patients likely to benefit from immunomodulatory therapies. It also appears reasonable to look for new therapeutic targets within this circuit. Published on behalf of the European Society of Cardiology. All rights reserved.
Authors: Rhian M Touyz; Marcus O E Boyd; Tomasz Guzik; Sandosh Padmanabhan; Linsay McCallum; Christian Delles; Patrick B Mark; John R Petrie; Francisco Rios; Augusto C Montezano; Robert Sykes; Colin Berry Journal: CJC Open Date: 2021-06-16