scholarly journals Microanatomy of the Human Atherosclerotic Plaque by Single-Cell Transcriptomics

2020 ◽  
Vol 127 (11) ◽  
pp. 1437-1455 ◽  
Author(s):  
Marie A.C. Depuydt ◽  
Koen H.M. Prange ◽  
Lotte Slenders ◽  
Tiit Örd ◽  
Danny Elbersen ◽  
...  
Keyword(s):  
T Cells ◽  
Smooth Muscle ◽  
Smooth Muscle Cells ◽  
Single Cell ◽  
Association Studies ◽  
Myeloid Cells ◽  
Muscle Cells ◽  
Atherosclerotic Plaques ◽  
Genome Wide Association Studies ◽  
Mesenchymal Transition

Rationale: Atherosclerotic lesions are known for their cellular heterogeneity, yet the molecular complexity within the cells of human plaques has not been fully assessed. Objective: Using single-cell transcriptomics and chromatin accessibility, we gained a better understanding of the pathophysiology underlying human atherosclerosis. Methods and Results: We performed single-cell RNA and single-cell ATAC sequencing on human carotid atherosclerotic plaques to define the cells at play and determine their transcriptomic and epigenomic characteristics. We identified 14 distinct cell populations including endothelial cells, smooth muscle cells, mast cells, B cells, myeloid cells, and T cells and identified multiple cellular activation states and suggested cellular interconversions. Within the endothelial cell population, we defined subsets with angiogenic capacity plus clear signs of endothelial to mesenchymal transition. CD4 + and CD8 + T cells showed activation-based subclasses, each with a gradual decline from a cytotoxic to a more quiescent phenotype. Myeloid cells included 2 populations of proinflammatory macrophages showing IL (interleukin) 1B or TNF (tumor necrosis factor) expression as well as a foam cell-like population expressing TREM2 (triggering receptor expressed on myeloid cells 2) and displaying a fibrosis-promoting phenotype. ATACseq data identified specific transcription factors associated with the myeloid subpopulation and T cell cytokine profiles underlying mutual activation between both cell types. Finally, cardiovascular disease susceptibility genes identified using public genome-wide association studies data were particularly enriched in lesional macrophages, endothelial, and smooth muscle cells. Conclusions: This study provides a transcriptome-based cellular landscape of human atherosclerotic plaques and highlights cellular plasticity and intercellular communication at the site of disease. This detailed definition of cell communities at play in atherosclerosis will facilitate cell-based mapping of novel interventional targets with direct functional relevance for the treatment of human disease.

Abstract 14092: Reduction of Atherosclerosis in Gucy1a3 Deficient Mice

Circulation ◽  
2015 ◽  
Vol 132 (suppl_3) ◽  
Author(s):  
Maria Segura Puimedon ◽  
Evanthia Mergia ◽  
Jaafar Al-Hasani ◽  
Redouane Aherrahrou ◽  
Jennifer Freyer ◽  
...  
Keyword(s):  
Blood Pressure ◽  
Smooth Muscle ◽  
Smooth Muscle Cells ◽  
Association Studies ◽  
Muscle Cells ◽  
Independent Experiment ◽  
Standard Diet ◽  
Genome Wide Association Studies ◽  
Primary Mouse

Introduction: GUCY1A3 encodes the α1 subunit of the soluble guanylate cyclase (sGC) and has been associated with CAD/MI by genome-wide association studies (GWAS) and in a rare extended family with MI. The α1 and the β1 subunit form the heterodimeric sGC, the nitric oxide (NO) receptor. NO plays an important role in the human cardiovascular system. The aim of our study is to define the role of GUCY1A3 in atherosclerosis using a mouse atherogenic model. Methods: Gucy1a3 KO mice were backcrossed on an Ldlr KO atherogenic background. Four groups were used, i.e. C57BL/6 (WT), Gucy1a3 KO, Ldlr KO and Gucy1a3-Ldlr double KO. All groups were fed high fat diet (HFD) or standard diet (SD) starting at the age of 12-14w and for 20 weeks. Blood pressure was studied at the end of the experiment in the HFD. At the start and end of the diet, lipid metabolism parameters (TC, HDLC, LDLC and TG) were analyzed. Plaque lesion size and collagen and macrophages content were studied. As smooth muscle cells play a key role in atherosclerosis, primary mouse aortic smooth muscle cells (maSMC) were isolated from WT and Gucy1a3 KO animals and subjected to proliferation and migration assays. Results: Blood pressure was studied in HFD animals, showing, as expected, an increase of systolic and diastolic blood pressure in double KO animals when compared to Ldlr KO animals. Regarding lipid levels, no differences were found in the double KO to Ldlr KO comparison under HFD, but a reduction in total cholesterol was found in double KO under SD. For atherosclerosis, a significant reduction in plaque area at the aortic root of double KO when compared to the Ldlr KO animals was demonstrated in both SD and HFD. The reduction was further confirmed in an additional independent experiment under SD. Similarly, a significant reduction of plaque lesion was found in the whole aorta of double KO when compared to the Ldlr KO under HFD. Moreover, a reduction in collagen content was found in double KO mice under HFD when compared to Ldlr KO animals. The migration and proliferation assays show a significant reduction in both migration and proliferation of the Gucy1a3 KO maSMC compared to WT. Conclusions: Our results point to an atherogenic role of Gucy1a3 in mice probably induced via smooth muscle migration and proliferation defects.

scholarly journals Intersecting single-cell transcriptomics and genome-wide association studies identifies crucial cell populations and candidate genes for atherosclerosis.

2021 ◽  
Author(s):  
Lotte Slenders ◽  
Lennart P.L. Landsmeer ◽  
Kai Cui ◽  
Marie A.C. Depuydt ◽  
Maarten Verwer ◽  
...  
Keyword(s):  
Smooth Muscle ◽  
Single Cell ◽  
Candidate Genes ◽  
Association Studies ◽  
Genome Wide Association ◽  
Atherosclerotic Plaques ◽  
Genome Wide Association Studies ◽  
Biological Mechanisms ◽  
Genome Wide ◽  
Artery Disease

Background: Genome-wide association studies have discovered hundreds of common genetic variants for atherosclerotic disease and cardiovascular risk factors. The translation of susceptibility loci into biological mechanisms and targets for drug discovery remains challenging. Intersecting genetic and gene expression data has led to the identification of candidate genes. However, previously studied tissues are often non-diseased and heterogeneous in cell composition, hindering accurate candidate prioritization. Therefore, we analyzed single-cell transcriptomics from atherosclerotic plaques for cell-type-specific expression to identify atherosclerosis-associated candidate gene-cell pairs. Methods and Results: To identify disease-associated genes, we applied gene-based analyses using GWAS summary statistics from 46 atherosclerotic and cardiovascular disease, risk factors, and other traits. We then intersected these candidates with scRNA-seq data to identify genes specific for individual cell (sub)populations in atherosclerotic plaques. The coronary artery disease loci demonstrated a prominent signal in plaque smooth muscle cells (SKI, KANK2, SORT1) p-adj. = 0.0012, and endothelial cells (SLC44A1, ATP2B1) p-adj. = 0.0011. Further sub clustering revealed genes in risk loci for coronary calcification specifically enriched in a synthetic smooth muscle cell population. Finally, we used liver-derived scRNA-seq data and showed hepatocyte-specific enrichment of genes involved in serum lipid levels. Conclusion: We discovered novel gene-cell pairs, on top of known pairs, pointing to new biological mechanisms of atherosclerotic disease. We highlight that loci associated with coronary artery disease reveal prominent association levels in mainly plaque smooth muscle and endothelial cell populations. We present an intuitive single-cell transcriptomics-driven workflow rooted in human large-scale genetic studies to identify putative candidate genes and affected cells associated with cardiovascular traits. Collectively, our workflow allows for the identification of cell-specific targets relevant for atherosclerosis and can be universally applied to other complex genetic diseases and traits.

Epicardial Cells of Human Adults Can Undergo an Epithelial-to-Mesenchymal Transition and Obtain Characteristics of Smooth Muscle Cells In Vitro

Stem Cells ◽  
2007 ◽  
Vol 25 (2) ◽  
pp. 271-278 ◽  
Author(s):  
John van Tuyn ◽  
Douwe E. Atsma ◽  
Elizabeth M. Winter ◽  
Ietje van der Velde-van Dijke ◽  
Daniel A. Pijnappels ◽  
...  
Keyword(s):  
Smooth Muscle ◽  
Smooth Muscle Cells ◽  
Epithelial To Mesenchymal Transition ◽  
Muscle Cells ◽  
Mesenchymal Transition ◽  
Epicardial Cells ◽  
Human Adults

scholarly journals Nuclear factor of activated T cells mediates oxidised LDL-induced calcification of vascular smooth muscle cells

Diabetologia ◽  
2011 ◽  
Vol 54 (10) ◽  
pp. 2690-2701 ◽  
Author(s):  
C. Goettsch ◽  
M. Rauner ◽  
C. Hamann ◽  
K. Sinningen ◽  
U. Hempel ◽  
...  
Keyword(s):  
T Cells ◽  
Smooth Muscle ◽  
Vascular Smooth Muscle ◽  
Smooth Muscle Cells ◽  
Vascular Smooth Muscle Cells ◽  
Muscle Cells ◽  
Nuclear Factor ◽  
Activated T Cells ◽  
Oxidised Ldl

scholarly journals Maintenance of AMPK activation by metformin is beneficial for suppressing the progress of diabetes-accelerated atherosclerosis via inhibition of vascular smooth muscle cells migration

2020 ◽  
Author(s):  
Yi Yan ◽  
Ting Li ◽  
Zhonghao Li ◽  
Mingyuan He ◽  
Dejiang Wang ◽  
...  
Keyword(s):  
Smooth Muscle ◽  
Vascular Smooth Muscle ◽  
Smooth Muscle Cells ◽  
Vascular Smooth Muscle Cells ◽  
High Fat Diet ◽  
Muscle Cells ◽  
Atherosclerotic Plaques ◽  
Ampk Activation ◽  
High Fat ◽  
Accelerated Atherosclerosis

Abstract Background: Our previous work revealed that augmented AMPK activation inhibit cell migration by phosphorylating its substrate Pdlim5. As medial VSMCs contribute to the major composition of atherosclerotic plaques, a hypothesis is raised that modulation of AMPK-Pdlim5 signal pathway could retard the development of atherosclerosis through inhibiting migration of VSMCs. Therefore, we initiate the present study to investigate whether AMPK agonist like metformin is beneficial for suppressing diabetes-accelerated atherosclerosis in a diabetic mouse model induced by streptozotocin and high fat diet.Methods: For cell experiment, vascular smooth muscle cells (VSMCs) were overexpressed flag fused Pdlim5 and Pdlim5 mutant. Then the engineered VSMCs were introduced with metformin or control drug before determination of phosphorylated Pdlim5 with immunoblotting. For animal work, 8-week-old male ApoE−/−mice were induced diabetes with streptozotocin and then were randomly divided into 8 groups: control group, metformin hydrochloride (300 mg/kg/day) group, wildtype-Pdlim5 (Pdlim5 WT) carried adenovirus (Ad) group, Ad Pdlim5 WT and Met group, Ad Pdlim5 S177A group, Ad Pdlim5 S177A and Met group, Ad Pdlim5 S177D group, Ad Pdlim5 S177D and Met group. All mice were fed with high fat diet after virus infection. At the end, mice were sacrificed to observe atherosclerotic plaques and deposition of VSMCs in plaques. Moreover, 12–15-week-old Myh11-cre-EGFP male mice were accepted ligation of the left carotid artery and randomly divided into control and metformin treatment group. Finally, the injured vessel of Myh11-cre-EGFP mice were isolated to analyze the relationship between AMPK activation and neointima formation.Results: It was found that AMPK directly phosphorylate Pdlim5 at Ser177 in vitro, and metformin, an AMPK agonist, could induce phosphorylation of Pdlim5 indirectly and inhibition of cell migration as a result. Exogenous expression of phosphomimetic S177D-Pdlim5 inhibits lamellipodia formation and migration in VSMCs. It was also demonstrated that VSMCs contribute to the major composition of injury-induced neointimal lesions, while metformin could alleviate the occlusion of carotid artery in a wire-injury mice model. In order to investigate the function of AMPK-Pdlim5 pathway in the context of pathological condition, ApoE−/− male mice were divided randomly into control, streptozocin and high fat diet-induced diabetes mellitus, STZ + HFD together with metformin or Pdlim5 mutant carried adenovirus treatment groups. The results showed increased plasma lipids and aggravated vascular smooth muscle cells infiltration into the atherosclerotic lesion in diabetic mice compared with control mice. However, metformin alleviated diabetes-induced metabolic disorders and atherosclerosis, as well as decreased VSMCs infiltration in atherosclerotic plaques, while Pdlim5 phospho-abolished mutant carried adenovirus S177A-Pdlim5 undermine this protective function.Conclusions: The activation of AMPK-Pdlim5 pathway by chemicals like Metformin could inhibit formation of migratory machine of VSMCs and alleviate the progress of atherosclerotic plaques in diabetic mice. The maintenance of AMPK activity is beneficial for suppressing diabetes-accelerated atherosclerosis or metabolic syndrome.

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