Eva Karlöf1, Till Seime2, Nuno Dias3, Mariette Lengquist2, Anna Witasp4, Håkan Almqvist5, Malin Kronqvist2, Jesper R Gådin6, Jacob Odeberg7, Lars Maegdefessel8, Peter Stenvinkel4, Ljubica Perisic Matic9, Ulf Hedin10. 1. Department of Vascular Surgery, Karolinska University Hospital, Stockholm, Sweden; Department of Molecular Medicine and Surgery, Karolinska Institutet, Stockholm, Sweden. 2. Department of Molecular Medicine and Surgery, Karolinska Institutet, Stockholm, Sweden. 3. Vascular Center, Department of Vascular Surgery, Skåne University Hospital, Malmö, Sweden. 4. Division of Renal Medicine, Department of Clinical Sciences, Intervention and Technology, Karolinska Institutet, Stockholm, Sweden. 5. Department of Clinical Neuroscience, Karolinska Institutet, Stockholm, Sweden. 6. Department of Medicine, Center for Molecular Medicine, Karolinska Institutet, Stockholm, Sweden. 7. Science for Life Laboratory, Department of Proteomics, School of Biotechnology, Royal Institute of Technology, Stockholm, Sweden. 8. Department of Medicine, Center for Molecular Medicine, Karolinska Institutet, Stockholm, Sweden; Department of Vascular and Endovascular Surgery, Klinikum Klinikum rechts der Isar Isar, Technical University Munich, Munich, Germany. 9. Department of Molecular Medicine and Surgery, Karolinska Institutet, Stockholm, Sweden. Electronic address: Ljubica.Matic@ki.se. 10. Department of Vascular Surgery, Karolinska University Hospital, Stockholm, Sweden; Department of Molecular Medicine and Surgery, Karolinska Institutet, Stockholm, Sweden. Electronic address: Ulf.Hedin@ki.se.
Abstract
BACKGROUND AND AIMS: Unstable carotid atherosclerosis causes stroke, but methods to identify patients and lesions at risk are lacking. We recently found enrichment of genes associated with calcification in carotid plaques from asymptomatic patients. Here, we hypothesized that calcification represents a stabilising feature of plaques and investigated how macro-calcification, as estimated by computed tomography (CT), correlates with gene expression profiles in lesions. METHODS: Plaque calcification was measured in pre-operative CT angiographies. Plaques were sorted into high- and low-calcified, profiled with microarrays, followed by bioinformatic analyses. Immunohistochemistry and qPCR were performed to evaluate the findings in plaques and arteries with medial calcification from chronic kidney disease patients. RESULTS: Smooth muscle cell (SMC) markers were upregulated in high-calcified plaques and calcified plaques from symptomatic patients, whereas macrophage markers were downregulated. The most enriched processes in high-calcified plaques were related to SMCs and extracellular matrix (ECM) organization, while inflammation, lipid transport and chemokine signaling were repressed. These findings were confirmed in arteries with high medial calcification. Proteoglycan 4 (PRG4) was identified as the most upregulated gene in association with plaque calcification and found in the ECM, SMA+ and CD68+/TRAP + cells. CONCLUSIONS: Macro-calcification in carotid lesions correlated with a transcriptional profile typical for stable plaques, with altered SMC phenotype and ECM composition and repressed inflammation. PRG4, previously not described in atherosclerosis, was enriched in the calcified ECM and localized to activated macrophages and smooth muscle-like cells. This study strengthens the notion that assessment of calcification may aid evaluation of plaque phenotype and stroke risk.
BACKGROUND AND AIMS: Unstable carotid atherosclerosis causes stroke, but methods to identify patients and lesions at risk are lacking. We recently found enrichment of genes associated with calcification in carotid plaques from asymptomatic patients. Here, we hypothesized that calcification represents a stabilising feature of plaques and investigated how macro-calcification, as estimated by computed tomography (CT), correlates with gene expression profiles in lesions. METHODS: Plaque calcification was measured in pre-operative CT angiographies. Plaques were sorted into high- and low-calcified, profiled with microarrays, followed by bioinformatic analyses. Immunohistochemistry and qPCR were performed to evaluate the findings in plaques and arteries with medial calcification from chronic kidney diseasepatients. RESULTS: Smooth muscle cell (SMC) markers were upregulated in high-calcified plaques and calcified plaques from symptomatic patients, whereas macrophage markers were downregulated. The most enriched processes in high-calcified plaques were related to SMCs and extracellular matrix (ECM) organization, while inflammation, lipid transport and chemokine signaling were repressed. These findings were confirmed in arteries with high medial calcification. Proteoglycan 4 (PRG4) was identified as the most upregulated gene in association with plaque calcification and found in the ECM, SMA+ and CD68+/TRAP + cells. CONCLUSIONS:Macro-calcification in carotid lesions correlated with a transcriptional profile typical for stable plaques, with altered SMC phenotype and ECM composition and repressed inflammation. PRG4, previously not described in atherosclerosis, was enriched in the calcified ECM and localized to activated macrophages and smooth muscle-like cells. This study strengthens the notion that assessment of calcification may aid evaluation of plaque phenotype and stroke risk.
Authors: Redouane Aherrahrou; Liang Guo; V Peter Nagraj; Aaron Aguhob; Jameson Hinkle; Lisa Chen; Joon Yuhl Soh; Dillon Lue; Gabriel F Alencar; Arjan Boltjes; Sander W van der Laan; Emily Farber; Daniela Fuller; Rita Anane-Wae; Ngozi Akingbesote; Ani W Manichaikul; Lijiang Ma; Minna U Kaikkonen; Johan L M Björkegren; Suna Önengüt-Gümüşcü; Gerard Pasterkamp; Clint L Miller; Gary K Owens; Aloke Finn; Mohamad Navab; Alan M Fogelman; Judith A Berliner; Mete Civelek Journal: Circ Res Date: 2020-10-12 Impact factor: 17.367
Authors: Gonzalo Artiach; Miguel Carracedo; Till Seime; Oscar Plunde; Andres Laguna-Fernandez; Ljubica Matic; Anders Franco-Cereceda; Magnus Bäck Journal: Cells Date: 2020-03-11 Impact factor: 6.600
Authors: Mirjami Laivuori; Johanna Tolva; A Inkeri Lokki; Nina Linder; Johan Lundin; Riitta Paakkanen; Anders Albäck; Maarit Venermo; Mikko I Mäyränpää; Marja-Liisa Lokki; Juha Sinisalo Journal: Front Cardiovasc Med Date: 2020-12-10
Authors: Till Seime; Asim Cengiz Akbulut; Moritz Lindquist Liljeqvist; Antti Siika; Hong Jin; Greg Winski; Rick H van Gorp; Eva Karlöf; Mariette Lengquist; Andrew J Buckler; Malin Kronqvist; Olivia J Waring; Jan H N Lindeman; Erik A L Biessen; Lars Maegdefessel; Anton Razuvaev; Leon J Schurgers; Ulf Hedin; Ljubica Matic Journal: Cells Date: 2021-05-21 Impact factor: 6.600
Authors: L Saba; W Brinjikji; J D Spence; M Wintermark; M Castillo; G J de Borst; Q Yang; C Yuan; A Buckler; M Edjlali; T Saam; D Saloner; B K Lal; D Capodanno; J Sun; N Balu; R Naylor; A V D Lugt; B A Wasserman; M E Kooi; J Wardlaw; J Gillard; G Lanzino; U Hedin; D Mikulis; A Gupta; J K DeMarco; C Hess; J V Goethem; T Hatsukami; P Rothwell; M M Brown; A R Moody Journal: AJNR Am J Neuroradiol Date: 2021-07-29 Impact factor: 4.966