Atheroprotective roles of smooth muscle cell phenotypic modulation and the TCF21 disease gene as revealed by single-cell analysis.
Animals
Basic Helix-Loop-Helix Transcription Factors
/ genetics
Cells, Cultured
Coronary Artery Disease
/ prevention & control
Humans
Mice
Mice, Inbred C57BL
Myocytes, Smooth Muscle
/ physiology
Osteoprotegerin
/ genetics
Phenotype
Polymorphism, Single Nucleotide
Sequence Analysis, RNA
Single-Cell Analysis
/ methods
Journal
Nature medicine
ISSN: 1546-170X
Titre abrégé: Nat Med
Pays: United States
ID NLM: 9502015
Informations de publication
Date de publication:
08 2019
08 2019
Historique:
received:
18
09
2018
accepted:
05
06
2019
pubmed:
31
7
2019
medline:
7
11
2019
entrez:
31
7
2019
Statut:
ppublish
Résumé
In response to various stimuli, vascular smooth muscle cells (SMCs) can de-differentiate, proliferate and migrate in a process known as phenotypic modulation. However, the phenotype of modulated SMCs in vivo during atherosclerosis and the influence of this process on coronary artery disease (CAD) risk have not been clearly established. Using single-cell RNA sequencing, we comprehensively characterized the transcriptomic phenotype of modulated SMCs in vivo in atherosclerotic lesions of both mouse and human arteries and found that these cells transform into unique fibroblast-like cells, termed 'fibromyocytes', rather than into a classical macrophage phenotype. SMC-specific knockout of TCF21-a causal CAD gene-markedly inhibited SMC phenotypic modulation in mice, leading to the presence of fewer fibromyocytes within lesions as well as within the protective fibrous cap of the lesions. Moreover, TCF21 expression was strongly associated with SMC phenotypic modulation in diseased human coronary arteries, and higher levels of TCF21 expression were associated with decreased CAD risk in human CAD-relevant tissues. These results establish a protective role for both TCF21 and SMC phenotypic modulation in this disease.
Identifiants
pubmed: 31359001
doi: 10.1038/s41591-019-0512-5
pii: 10.1038/s41591-019-0512-5
pmc: PMC7274198
mid: NIHMS1531197
doi:
Substances chimiques
Basic Helix-Loop-Helix Transcription Factors
0
Osteoprotegerin
0
TCF21 protein, human
0
TNFRSF11B protein, human
0
Types de publication
Journal Article
Research Support, N.I.H., Extramural
Langues
eng
Sous-ensembles de citation
IM
Pagination
1280-1289Subventions
Organisme : NHLBI NIH HHS
ID : R01 HL109512
Pays : United States
Organisme : NHLBI NIH HHS
ID : R01 HL144067
Pays : United States
Organisme : NIDDK NIH HHS
ID : R01 DK107437
Pays : United States
Organisme : NHLBI NIH HHS
ID : R01 HL139478
Pays : United States
Organisme : NHLBI NIH HHS
ID : K08 HL133375
Pays : United States
Organisme : NHLBI NIH HHS
ID : R01 HL103635
Pays : United States
Organisme : NIH HHS
ID : S10 OD018220
Pays : United States
Organisme : NCRR NIH HHS
ID : S10 RR025518
Pays : United States
Organisme : NHLBI NIH HHS
ID : R33 HL120757
Pays : United States
Organisme : NHLBI NIH HHS
ID : R21 HL120757
Pays : United States
Organisme : NHLBI NIH HHS
ID : F32 HL129670
Pays : United States
Organisme : NHLBI NIH HHS
ID : R01 HL148239
Pays : United States
Organisme : NHLBI NIH HHS
ID : R01 HL134817
Pays : United States
Organisme : NHLBI NIH HHS
ID : R01 HL141371
Pays : United States
Organisme : NHLBI NIH HHS
ID : R00 HL125912
Pays : United States
Organisme : NHLBI NIH HHS
ID : R01 HL145708
Pays : United States
Organisme : NIDDK NIH HHS
ID : P30 DK116074
Pays : United States
Références
Davies, M. J., Richardson, P. D., Woolf, N., Katz, D. R. & Mann, J. Risk of thrombosis in human atherosclerotic plaques: role of extracellular lipid, macrophage, and smooth muscle cell content. Br. Heart J. 69, 377–381 (1993).
doi: 10.1136/hrt.69.5.377
Libby, P. & Aikawa, M. Stabilization of atherosclerotic plaques: new mechanisms and clinical targets. Nat. Med. 8, 1257–1262 (2002).
doi: 10.1038/nm1102-1257
Ross, R. & Glomset, J. A. Atherosclerosis and the arterial smooth muscle cell: proliferation of smooth muscle is a key event in the genesis of the lesions of atherosclerosis. Science 180, 1332–1339 (1973).
doi: 10.1126/science.180.4093.1332
Bennett, M. R., Sinha, S. & Owens, G. K. Vascular smooth muscle cells in atherosclerosis. Circ. Res. 118, 692–702 (2016).
doi: 10.1161/CIRCRESAHA.115.306361
Gomez, D. & Owens, G. K. Smooth muscle cell phenotypic switching in atherosclerosis. Cardiovasc. Res. 95, 156–164 (2012).
doi: 10.1093/cvr/cvs115
Shankman, L. S. et al. KLF4-dependent phenotypic modulation of smooth muscle cells has a key role in atherosclerotic plaque pathogenesis. Nat. Med. 21, 628–637 (2015).
doi: 10.1038/nm.3866
Gomez, D., Shankman, L. S., Nguyen, A. T. & Owens, G. K. Detection of histone modifications at specific gene loci in single cells in histological sections. Nat. Methods 10, 171–177 (2013).
doi: 10.1038/nmeth.2332
Schunkert, H. et al. Large-scale association analysis identifies 13 new susceptibility loci for coronary artery disease. Nat. Genet. 43, 333–338 (2011).
doi: 10.1038/ng.784
Miller, C. L. et al. Disease-related growth factor and embryonic signaling pathways modulate an enhancer of TCF21 expression at the 6q23.2 coronary heart disease locus. PLoS Genet. 9, e1003652 (2013).
doi: 10.1371/journal.pgen.1003652
Miller, C. L. et al. Coronary heart disease-associated variation in TCF21 disrupts a miR-224 binding site and miRNA-mediated regulation. PLoS Genet. 10, e1004263 (2014).
doi: 10.1371/journal.pgen.1004263
Dettman, R. W., Denetclaw, W. Jr, Ordahl, C. P. & Bristow, J. Common epicardial origin of coronary vascular smooth muscle, perivascular fibroblasts, and intermyocardial fibroblasts in the avian heart. Dev. Biol. 193, 169–181 (1998).
doi: 10.1006/dbio.1997.8801
Winter, E. M. & Gittenberger-de Groot, A. C. Epicardium-derived cells in cardiogenesis and cardiac regeneration. Cell. Mol. Life Sci. 64, 692–703 (2007).
doi: 10.1007/s00018-007-6522-3
Acharya, A. et al. The bHLH transcription factor Tcf21 is required for lineage-specific EMT of cardiac fibroblast progenitors. Development 139, 2139–2149 (2012).
doi: 10.1242/dev.079970
Nurnberg, S. T. et al. Coronary artery disease associated transcription factor TCF21 regulates smooth muscle precursor cells that contribute to the fibrous cap. PLoS Genet. 11, e1005155 (2015).
doi: 10.1371/journal.pgen.1005155
Herring, B. P., Hoggatt, A. M., Burlak, C. & Offermanns, S. Previously differentiated medial vascular smooth muscle cells contribute to neointima formation following vascular injury. Vasc. Cell 6, 21 (2014).
doi: 10.1186/2045-824X-6-21
Wirth, A. et al. G12-G13-LARG-mediated signaling in vascular smooth muscle is required for salt-induced hypertension. Nat. Med. 14, 64–68 (2008).
doi: 10.1038/nm1666
Madisen, L. et al. A robust and high-throughput Cre reporting and characterization system for the whole mouse brain. Nat. Neurosci. 13, 133–140 (2010).
doi: 10.1038/nn.2467
Zheng, G. X. et al. Massively parallel digital transcriptional profiling of single cells. Nat. Commun. 8, 14049 (2017).
doi: 10.1038/ncomms14049
Dobnikar, L. et al. Disease-relevant transcriptional signatures identified in individual smooth muscle cells from healthy mouse vessels. Nat. Commun. 9, 4567 (2018).
doi: 10.1038/s41467-018-06891-x
Kitchen, C. M., Cowan, S. L., Long, X. & Miano, J. M. Expression and promoter analysis of a highly restricted integrin alpha gene in vascular smooth muscle. Gene 513, 82–89 (2013).
doi: 10.1016/j.gene.2012.10.073
Stoeckius, M. et al. Simultaneous epitope and transcriptome measurement in single cells. Nat. Methods 14, 865–868 (2017).
doi: 10.1038/nmeth.4380
Jacobsen, K. et al. Diverse cellular architecture of atherosclerotic plaque derives from clonal expansion of a few medial SMCs. JCI Insight 2, 95890 (2017).
doi: 10.1172/jci.insight.95890
Vengrenyuk, Y. et al. Cholesterol loading reprograms the microRNA-143/145–myocardin axis to convert aortic smooth muscle cells to a dysfunctional macrophage-like phenotype. Arterioscler. Thromb. Vasc. Biol. 35, 535–546 (2015).
doi: 10.1161/ATVBAHA.114.304029
Sazonova, O. et al. Characterization of TCF21 downstream target regions identifies a transcriptional network linking multiple independent coronary artery disease loci. PLoS Genet. 11, e1005202 (2015).
doi: 10.1371/journal.pgen.1005202
Franzen, O. et al. Cardiometabolic risk loci share downstream cis- and trans-gene regulation across tissues and diseases. Science 353, 827–830 (2016).
doi: 10.1126/science.aad6970
Clement, N. et al. Notch3 and IL-1β exert opposing effects on a vascular smooth muscle cell inflammatory pathway in which NF-κB drives crosstalk. J. Cell Sci. 120, 3352–3361 (2007).
doi: 10.1242/jcs.007872
Pidkovka, N. A. et al. Oxidized phospholipids induce phenotypic switching of vascular smooth muscle cells in vivo and in vitro. Circ. Res. 101, 792–801 (2007).
doi: 10.1161/CIRCRESAHA.107.152736
Dandre, F. & Owens, G. K. Platelet-derived growth factor-BB and Ets-1 transcription factor negatively regulate transcription of multiple smooth muscle cell differentiation marker genes. Am. J. Physiol. Heart Circ. Physiol. 286, H2042–H2051 (2004).
doi: 10.1152/ajpheart.00625.2003
Rong, J. X., Shapiro, M., Trogan, E. & Fisher, E. A. Transdifferentiation of mouse aortic smooth muscle cells to a macrophage-like state after cholesterol loading. Proc. Natl Acad. Sci. USA 100, 13531–13536 (2003).
doi: 10.1073/pnas.1735526100
Rong, J. X., Berman, J. W., Taubman, M. B. & Fisher, E. A. Lysophosphatidylcholine stimulates monocyte chemoattractant protein-1 gene expression in rat aortic smooth muscle cells. Arterioscler. Thromb. Vasc. Biol. 22, 1617–1623 (2002).
doi: 10.1161/01.ATV.0000035408.93749.71
Allahverdian, S., Chehroudi, A. C., McManus, B. M., Abraham, T. & Francis, G. A. Contribution of intimal smooth muscle cells to cholesterol accumulation and macrophage-like cells in human atherosclerosis. Circulation 129, 1551–1559 (2014).
doi: 10.1161/CIRCULATIONAHA.113.005015
Kanisicak, O. et al. Genetic lineage tracing defines myofibroblast origin and function in the injured heart. Nat. Commun. 7, 12260 (2016).
doi: 10.1038/ncomms12260
Chappell, J. et al. Extensive proliferation of a subset of differentiated, yet plastic, medial vascular smooth muscle cells contributes to neointimal formation in mouse injury and atherosclerosis models. Circ. Res. 119, 1313–1323 (2016).
doi: 10.1161/CIRCRESAHA.116.309799
Butler, A., Hoffman, P., Smibert, P., Papalexi, E. & Satija, R. Integrating single-cell transcriptomic data across different conditions, technologies, and species. Nat. Biotechnol. 36, 411–420 (2018).
doi: 10.1038/nbt.4096
Satija, R., Farrell, J. A., Gennert, D., Schier, A. F. & Regev, A. Spatial reconstruction of single-cell gene expression data. Nat. Biotechnol. 33, 495–502 (2015).
doi: 10.1038/nbt.3192
Kasowski, M. et al. Variation in transcription factor binding among humans. Science 328, 232–235 (2010).
doi: 10.1126/science.1183621
Li, H. & Durbin, R. Fast and accurate short read alignment with Burrows–Wheeler transform. Bioinformatics 25, 1754–1760 (2009).
doi: 10.1093/bioinformatics/btp324
Zhang, Y. et al. Model-based analysis of ChIP-Seq (MACS). Genome Biol. 9, R137 (2008).
doi: 10.1186/gb-2008-9-9-r137