Leukocytes carrying Clonal Hematopoiesis of Indeterminate Potential (CHIP) Mutations invade Human Atherosclerotic Plaques

medRxiv [Preprint]. 2023 Jul 26:2023.07.22.23292754. doi: 10.1101/2023.07.22.23292754.

Authors

Moritz von Scheidt^{1

2}, Sabine Bauer^{1

2}, Angela Ma³, Ke Hao³, Thorsten Kessler^{1

2}, Baiba Vilne^{2

4

5}, Ying Wang⁶, Chani J Hodonsky⁷, Saikat K B Ghosh⁸, Michal Mokry^{9

10}, Hua Gao¹¹, Kenji Kawai⁸, Atsushi Sakamoto^{8

12}, Juliane Kaiser¹³, Dario Bongiovanni^{14

15}, Julia Fleig¹, Lilith Oldenbuettel¹, Zhifen Chen^{1

2}, Aldo Moggio^{1

2}, Hendrik B Sager^{1

2}, Judith S Hecker¹⁶, Florian Bassermann¹⁶, Lars Maegdefessel^{2

17}, Clint L Miller¹⁸, Wolfgang Koenig^{1

2}, Andreas M Zeiher¹⁹, Stefanie Dimmeler¹⁹, Matthias Graw¹³, Christian Braun¹³, Arno Ruusalepp^{20

21

22}, Nicholas J Leeper^{10

23}, Jason C Kovacic^{24

25

26}, Johan L M Björkegren^{3

21

27}, Heribert Schunkert^{1

2}

Affiliations

¹ Department of Cardiology, German Heart Center Munich, Technical University Munich, Munich, Germany.
² Deutsches Zentrum für Herz- und Kreislaufforschung (DZHK), Partner Site Munich Heart Alliance, Munich, Germany.
³ Department of Genetics and Genomic Sciences, Institute of Genomics and Multiscale Biology, Icahn School of Medicine at Mount Sinai, New York, USA.
⁴ Bioinformatics Lab, Riga Stradiņš University, Riga, Latvia.
⁵ SIA Net-OMICS, Riga, Latvia.
⁶ Department of Pathology and Laboratory Medicine, Centre for Heart Lung Innovation, University of British Columbia, Vancouver, Canada.
⁷ Center for Public Health Genomics, University of Virginia, Charlottesville, VA, USA.
⁸ CVPath Institute, Inc, Gaithersburg, USA.
⁹ Laboratory of Experimental Cardiology, Department of Cardiology, University Medical Center Utrecht, University Utrecht, Utrecht, Netherlands.
¹⁰ Central Diagnostics Laboratory, University Medical Center Utrecht, Utrecht, The Netherlands.
¹¹ Division of Vascular Surgery, Department of Surgery, Stanford University School of Medicine, Stanford, USA.
¹² Division of Cardiology, Internal Medicine III, Hamamatsu University School of Medicine, Hamamatsu, Japan.
¹³ Institute of Legal Medicine, Faculty of Medicine, LMU Munich, Germany.
¹⁴ Department of Internal Medicine I, Cardiology, University Hospital Augsburg, University of Augsburg, Germany.
¹⁵ Department of Cardiovascular Medicine, Humanitas Clinical and Research Center IRCCS and Humanitas University, Rozzano, Milan, Italy.
¹⁶ Department of Medicine III, Technical University of Munich (TUM), Klinikum rechts der Isar, Munich, Germany.
¹⁷ Department for Vascular and Endovascular Surgery, Klinikum Rechts der Isar, Technical University Munich, Munich, Germany.
¹⁸ Center for Public Health Genomics, Department of Public Health Sciences, Department of Biochemistry and Molecular Genetics, University of Virginia, Charlottesville, VA, USA.
¹⁹ Institute for Cardiovascular Regeneration, Goethe University Frankfurt am Main, Frankfurt am Main, Germany.
²⁰ Department of Cardiac Surgery, The Heart Clinic, Tartu University Hospital, Tartu, Estonia.
²¹ Clinical Gene Networks AB, Stockholm, Sweden.
²² Institute of Clinical Medicine, Faculty of Medicine, Tartu University, Tartu, Estonia.
²³ Stanford Cardiovascular Institute, Stanford University, Stanford, USA.
²⁴ Victor Chang Cardiac Research Institute, Darlinghurst, Australia.
²⁵ St. Vincent's Clinical School, University of New South Wales, Sydney, Australia.
²⁶ Cardiovascular Research Institute, Icahn School of Medicine at Mount Sinai, New York, USA.
²⁷ Department of Medicine, Huddinge, Karolinska Institutet, Karolinska Universitetssjukhuset, Stockholm, Sweden.

Abstract

Background: Leukocyte progenitors derived from clonal hematopoiesis of undetermined potential (CHIP) are associated with increased cardiovascular events. However, the prevalence and functional relevance of CHIP in coronary artery disease (CAD) are unclear, and cells affected by CHIP have not been detected in human atherosclerotic plaques.

Methods: CHIP mutations in blood and tissues were identified by targeted deep-DNA-sequencing (DNAseq: coverage >3,000) and whole-genome-sequencing (WGS: coverage >35). CHIP-mutated leukocytes were visualized in human atherosclerotic plaques by mutaFISH^™. Functional relevance of CHIP mutations was studied by RNAseq.

Results: DNAseq of whole blood from 540 deceased CAD patients of the Munich cardIovaScular StudIes biObaNk (MISSION) identified 253 (46.9%) CHIP mutation carriers (mean age 78.3 years). DNAseq on myocardium, atherosclerotic coronary and carotid arteries detected identical CHIP mutations in 18 out of 25 mutation carriers in tissue DNA. MutaFISH^™ visualized individual macrophages carrying DNMT3A CHIP mutations in human atherosclerotic plaques. Studying monocyte-derived macrophages from Stockholm-Tartu Atherosclerosis Reverse Networks Engineering Task (STARNET; n=941) by WGS revealed CHIP mutations in 14.2% (mean age 67.1 years). RNAseq of these macrophages revealed that expression patterns in CHIP mutation carriers differed substantially from those of non-carriers. Moreover, patterns were different depending on the underlying mutations, e.g. those carrying TET2 mutations predominantly displayed upregulated inflammatory signaling whereas ASXL1 mutations showed stronger effects on metabolic pathways.

Conclusions: Deep-DNA-sequencing reveals a high prevalence of CHIP mutations in whole blood of CAD patients. CHIP-affected leukocytes invade plaques in human coronary arteries. RNAseq data obtained from macrophages of CHIP-affected patients suggest that pro-atherosclerotic signaling differs depending on the underlying mutations. Further studies are necessary to understand whether specific pathways affected by CHIP mutations may be targeted for personalized treatment.

Keywords: aging; atherosclerotic cardiovascular disease; clonal hematopoiesis of indeterminate potential; coronary artery disease; inflammation.

Publication types

Preprint

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