Microregional RNA sequencing reveals layer-specific smooth muscle cell heterogeneity in human bicuspid aortic valve-associated aortic dilatation.
10 affiliations
- 1Life Science Center for Survival Dynamics, TARA, University of Tsukuba, Tsukuba, Japan.
- 2Division of Glocal Pharmacology, Department of Pharmacology, Graduate School of Medicine, The University of Osaka, Osaka, Japan.
- 3Doctoral Program in Medical Sciences, Graduate School of Comprehensive Human Sciences, University of Tsukuba, Tsukuba, Japan.
- 4Department of Cardiovascular Surgery, Saitama Medical Center, Saitama Medical University, Saitama, Japan.
- 5Research Organization for Nano and Life Innovation, Waseda University, Tokyo, Japan.
- 6Center for Spatial Omics Analysis Research, Waseda University, Tokyo, Japan.
- 7Division of Bioinformation and Therapeutic Systems, National Defense Medical College Research Institute, Saitama, Japan.
- 8Graduate School of Advanced Science and Engineering, Waseda University, Tokyo, Japan.
- 9Department of Cardiovascular Surgery, University of Tsukuba, Tsukuba, Japan.
- 10Faculty of Medicine, University of Tsukuba, Tsukuba, Japan.
Abstract
Objective
Smooth muscle cells in the human ascending aorta exhibit layer-specific heterogeneity; however, their molecular profiles and contribution to aortic diseases remain poorly understood. In this study, we applied microregional RNA sequencing to characterize layer-specific transcriptomic signatures and investigate the differences between bicuspid aortic valve (BAV) and tricuspid aortic valve (TAV) aortopathy.Methods
Dilated ascending aortic wall samples were obtained from nine patients who underwent ascending aorta replacement concomitant with aortic valve replacement: five with BAV and four with TAV. Microdissected tissues were collected from four distinct layers in media using a micro-punching device. RNA was extracted and sequenced, and the transcriptomic differences across the layers and between BAV- and TAV-dilated aortas were analyzed.Results
Microregional RNA sequencing revealed significant upregulation of several extracellular matrix remodeling genes in the innermost layer, including FN1, VCAN, MGP, and POSTN. Among them, POSTN was further validated at the protein level, confirming its upregulation in the innermost layer, supporting the validity of this analysis. Next, the transcriptional differences between BAV- and TAV-dilated aortas across individual layers were analyzed. The gene expression differences were the most prominent in the innermost layer. Gene set enrichment analysis revealed that epithelial-mesenchymal transition and transforming growth factor β signaling-related genes were enriched in BAV-dilated aortas. In contrast, inflammation-related genes were enriched in TAV-dilated aortas. We also evaluated the pathways associated with the genes implicated in BAV formation to identify potential upstream mechanisms, revealing that mechanosensing-related genes are coordinately suppressed in BAV-dilated aortas.Conclusions
The present study demonstrated the application of microregional RNA sequencing to human aortic tissue, revealing layer-specific transcriptomic differences in aortic diseases. The differences in gene-set-level findings indicate distinct biological contexts and mechanisms in BAV- and TAV-dilated aortas, particularly in the innermost media layer. This approach is useful for assessing layer-specific profiles in aortic diseases.Clinical relevance
This study identified layer-specific heterogeneity within the human aorta, with molecular alterations in the innermost medial layer potentially contributing to differences between TAV- and BAV-associated aortopathy. The findings suggest that layer-specific transcriptome profiles differ across disease types. Accordingly, diagnostic approaches and therapeutic strategies may be more effective when tailored to specific aortic regions and their underlying molecular characteristics.
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BibTeX
@article{koizumi2026microregional,
title = {{Microregional RNA sequencing reveals layer-specific smooth muscle cell heterogeneity in human bicuspid aortic valve-associated aortic dilatation.}},
author = {Shigeki Koizumi and Chiho Tokunaga and Keiichi Asano and Kenichi Kimura and Hiroko Matsunaga and Eri Motoyama and Hnin Htet Htet Aung and Kaori Sugiyama and Haruko Takeyama and Yuji Hiramatsu and Hiromi Yanagisawa},
journal = {JVS-vascular science},
year = {2026},
month = sep,
doi = {10.1016/j.jvssci.2026.100444},
url = {https://europepmc.org/article/MED/42908712},
note = {CC BY},
}
RIS
TY - JOUR
TI - Microregional RNA sequencing reveals layer-specific smooth muscle cell heterogeneity in human bicuspid aortic valve-associated aortic dilatation.
AU - Shigeki Koizumi
AU - Chiho Tokunaga
AU - Keiichi Asano
AU - Kenichi Kimura
AU - Hiroko Matsunaga
AU - Eri Motoyama
AU - Hnin Htet Htet Aung
AU - Kaori Sugiyama
AU - Haruko Takeyama
AU - Yuji Hiramatsu
AU - Hiromi Yanagisawa
PY - 2026
DA - 2026/09/16
JO - JVS-vascular science
DO - 10.1016/j.jvssci.2026.100444
UR - https://europepmc.org/article/MED/42908712
AB - Objective Smooth muscle cells in the human ascending aorta exhibit layer-specific heterogeneity; however, their molecular profiles and contribution to aortic diseases remain poorly understood. In this study, we applied microregional RNA sequencing to characterize layer-specific transcriptomic signatures and investigate the differences between bicuspid aortic valve (BAV) and tricuspid aortic valve (TAV) aortopathy.Methods Dilated ascending aortic wall samples were obtained from nine patients who underwent ascending aorta replacement concomitant with aortic valve replacement: five with BAV and four with TAV. Microdissected tissues were collected from four distinct layers in media using a micro-punching device. RNA was extracted and sequenced, and the transcriptomic differences across the layers and between BAV- and TAV-dilated aortas were analyzed.Results Microregional RNA sequencing revealed significant upregulation of several extracellular matrix remodeling genes in the innermost layer, including FN1, VCAN, MGP, and POSTN. Among them, POSTN was further validated at the protein level, confirming its upregulation in the innermost layer, supporting the validity of this analysis. Next, the transcriptional differences between BAV- and TAV-dilated aortas across individual layers were analyzed. The gene expression differences were the most prominent in the innermost layer. Gene set enrichment analysis revealed that epithelial-mesenchymal transition and transforming growth factor β signaling-related genes were enriched in BAV-dilated aortas. In contrast, inflammation-related genes were enriched in TAV-dilated aortas. We also evaluated the pathways associated with the genes implicated in BAV formation to identify potential upstream mechanisms, revealing that mechanosensing-related genes are coordinately suppressed in BAV-dilated aortas.Conclusions The present study demonstrated the application of microregional RNA sequencing to human aortic tissue, revealing layer-specific transcriptomic differences in aortic diseases. The differences in gene-set-level findings indicate distinct biological contexts and mechanisms in BAV- and TAV-dilated aortas, particularly in the innermost media layer. This approach is useful for assessing layer-specific profiles in aortic diseases.Clinical relevance This study identified layer-specific heterogeneity within the human aorta, with molecular alterations in the innermost medial layer potentially contributing to differences between TAV- and BAV-associated aortopathy. The findings suggest that layer-specific transcriptome profiles differ across disease types. Accordingly, diagnostic approaches and therapeutic strategies may be more effective when tailored to specific aortic regions and their underlying molecular characteristics.
SN - 2666-3503
ER -