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. 2025 Feb;45(2):e15-e29.
doi: 10.1161/ATVBAHA.124.321482. Epub 2025 Jan 2.

Single-Cell Transcriptomics Identifies Selective Lineage-Specific Regulation of Genes in Aortic Smooth Muscle Cells in Mice

Shalabh Shukla #  1 , Sayantan Jana #  1 , Nicole Sanford  1 , Chloe Y Lee  1 , Li Liu  1 , Paul Cheng  2 , Thomas Quertermous  2 , David A Dichek  1
Affiliations

Single-Cell Transcriptomics Identifies Selective Lineage-Specific Regulation of Genes in Aortic Smooth Muscle Cells in Mice

Shalabh Shukla et al. Arterioscler Thromb Vasc Biol. 2025 Feb.

Abstract

Background: Smooth muscle cells (SMCs) of the proximal thoracic aorta are derived from second heart field (SHF) and cardiac neural crest (CNC) lineages. Recent studies, both in vitro and in vivo, have implied relevance of lineage-specific SMC functions in the pathophysiology of thoracic aortic diseases; however, whether 2 lineage-derived SMCs have any predisposed transcriptional differences in the control aorta remains unexplored.

Methods: Single-cell RNA sequencing and single-nucleus assay for transposase-accessible chromatin sequencing were performed on isolated cells from the aortic root and ascending aortas of 14-week-old SHF-traced (Mef2c-Cre+/0-Yfp+/0) and CNC-traced (Wnt1-Cre+/0-Yfp+/0) male mice. RNA in situ hybridization was performed for spatial expression of selected differentially expressed genes (DEGs) of both lineages.

Results: Lineage stratification of SMCs in the proximal thoracic aorta was identified using antibody-based immunofluorescence staining. Single-cell RNA sequencing recognized 12 consistently upregulated DEGs (Des, Tnnt2, Hand2os1, Psd, Gpc3, Meis2, Dcn, Gm34030, Palld, Nrtn, Lum, and Cfh) in SHF-derived SMCs and 9 consistently upregulated DEGs (Ccn5, Ccdc42, Tes, Eln, Aebp1, Galnt6, Ccn2, Aopep, and Wtip) in CNC-derived SMCs. RNA in situ hybridization validated upregulated expressions of selective SHF-specific DEGs at the aortic root. We found SHF-derived SMCs contain a distinct, large subpopulation of SMCs that is enriched with Des and Tnnt2 expressions. Single-nucleus assay for transposase-accessible chromatin analysis further confirmed higher chromosomal accessibility for upregulated DEGs of SHF-derived SMCs.

Conclusions: The present study recognizes the presence of limited but distinct transcriptomic differences between CNC-derived and SHF-derived SMCs in the control proximal thoracic aorta.

Keywords: aorta; chromatin; gene expression profiling; neural crest.

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Conflict of interest statement

None.

References

    1. Majesky MW. Developmental basis of vascular smooth muscle diversity. Arterioscler Thromb Vasc Biol. 2007;27:1248–1258. doi: 10.1161/ATVBAHA.107.141069 - DOI - PubMed
    1. Sawada H, Rateri DL, Moorleghen JJ, Majesky MW, Daugherty A. Smooth Muscle Cells Derived From Second Heart Field and Cardiac Neural Crest Reside in Spatially Distinct Domains in the Media of the Ascending Aorta-Brief Report. Arterioscler Thromb Vasc Biol. 2017;37:1722–1726. doi: 10.1161/atvbaha.117.309599 - DOI - PMC - PubMed
    1. Fletcher AJ, Syed MBJ, Aitman TJ, Newby DE, Walker NL. Inherited Thoracic Aortic Disease: New Insights and Translational Targets. Circulation. 2020;141:1570–1587. doi: 10.1161/CIRCULATIONAHA.119.043756 - DOI - PMC - PubMed
    1. Loeys BL, Schwarze U, Holm T, Callewaert BL, Thomas GH, Pannu H, De Backer JF, Oswald GL, Symoens S, Manouvrier S, et al. Aneurysm syndromes caused by mutations in the TGF-beta receptor. N Engl J Med. 2006;355:788–798. doi: 10.1056/NEJMoa055695 - DOI - PubMed
    1. Roman MJ, Devereux RB, Kramer-Fox R, Spitzer MC. Comparison of cardiovascular and skeletal features of primary mitral valve prolapse and Marfan syndrome. Am J Cardiol. 1989;63:317–321. doi: 10.1016/0002-9149(89)90338-x - DOI - PubMed

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