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Updated: May 1, 2026
Isolation of Mouse Interstitial Valve Cells to Study the Calcification of the Aortic Valve In Vitro
Published on: May 10, 2021
Multiomics Analysis Reveals Stromal Cell State Changes and INHBA-Associated Remodeling in Calcific Aortic Valve
Shilin Jin1, Shishi Wu1, Feng Shen2
1Human Phenome Institute, Shanghai Pudong Hospital, Fudan University Pudong Medical Centre, Fudan University, China (S.J., S.W., C.M., M.S., D.Z., D.Y., M.Y.).
Insights
Researchers identified shifts in valvular interstitial cells (VICs) and valve-derived stromal cells (VDSCs) in calcific aortic valve disease (CAVD). The study implicates INHBA in VIC calcification and disease remodeling.
Area of Science:
- Cardiovascular Biology
- Developmental Biology
- Genomics
Background:
- Calcific aortic valve disease (CAVD) mechanisms are not fully understood.
- Signaling pathways active in valve development may play roles in CAVD.
- Context-specific roles of these pathways in CAVD require clarification.
Purpose of the Study:
- To identify CAVD-relevant cell populations and candidate genes using multiomics data.
- To investigate the role of INHBA in valvular interstitial cell (VIC) calcification.
- To analyze INHBA expression in in vivo CAVD models.
Main Methods:
- Integrated multiomics data: GWAS, single-cell RNA sequencing (scRNA-seq), bulk RNA sequencing.
- scRNA-seq analysis of fetal, healthy adult, and CAVD valves.
- In vitro VIC calcification model and in vivo CAVD model to assess INHBA function.
Main Results:
- Stage-associated changes in valve cell composition identified, with VDSC1 enrichment in CAVD.
- VDSC1 showed enrichment of inflammation and extracellular matrix remodeling pathways.
- INHBA expression increased during VIC osteogenic induction, and its silencing reduced calcification.
- 21 candidate CAVD genes identified, with INHBA showing stage-dependent expression.
Conclusions:
- VDSC1 enrichment is a key feature of CAVD, with stage-associated shifts in VIC and VDSC subpopulations.
- INHBA is implicated in VIC osteogenic remodeling.
- Further evaluation of INHBA in CAVD pathogenesis is warranted.
Background:
The cellular and molecular mechanisms underlying calcific aortic valve disease (CAVD) remain incompletely understood. Emerging evidence suggests that signaling components active during valve development may also be relevant to disease-associated remodeling, although their context-specific roles remain unclear.
Methods:
We integrated multiomics data, including genome-wide association study summary data, single-cell RNA sequencing from fetal, healthy adults, and CAVD valves, and bulk RNA-sequencing data sets to identify the CAVD-relevant cell populations and candidate genes. Then, an in vitro calcification model of valvular interstitial cells (VICs) was used to assess the role of INHBA (inhibin subunit βA) in VIC calcification and to examine its expression in an in vivo CAVD model.
Results:
Our single-cell analysis revealed stage-associated changes in valve cell composition across fetal, healthy, and CAVD conditions. VICs and valve-derived stromal cells (VDSCs) showed closely connected transcriptional states. Specifically, fetal VIC subpopulations, including VIC3 and VIC5, shifted toward adult VIC2 and VDSC2, and ultimately toward VDSC1-enriched states in CAVD. VDSC1 was consistently enriched in CAVD and showed enrichment of inflammation-related and extracellular matrix remodeling-related pathways. Cell-cell communication analysis showed significantly enhanced interactions between VDSC1 and macrophages in CAVD, consistent with increased inflammatory signaling in the local microenvironment. Integrated analysis of genome-wide association study, bulk RNA sequencing, and single-cell RNA sequencing identified 21 candidate genes associated with CAVD. Among these, INHBA showed stage-dependent expression across valve conditions. In vitro, INHBA expression increased during osteogenic induction of VICs, along with elevated RUNX2 (runt-related transcription factor 2) and ALP (alkaline phosphatase), whereas INHBA silencing reduced calcification. In vivo, INHBA expression also differed across fetal, healthy, and calcified valves.
Conclusions:
Those findings define stage-associated shifts in VIC and VDSC subpopulations across valve conditions and identify VDSC1 enrichment as a prominent feature of CAVD. Our data further implicate INHBA in osteogenic remodeling of VICs and support further evaluation of INHBA in disease-associated remodeling in CAVD.
