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Updated: Aug 15, 2026
Single Cell Transcriptional Profiling of Adult Mouse Cardiomyocytes
Published on: December 28, 2011
Integrative Single-Cell Analysis of Cardiomyopathy Identifies Differences in Cell Stemness and Transcriptional
Wenyang Nie1,2, Zhijie Zhao3,4, Yuhang Liu5
1Department of Cardiovascular Diseases, Affiliated Hospital of Shandong University of Traditional Chinese Medicine, 16369 Jing 10 Rd, Jinan 250000, China.
Insights
This study reveals that AGT+ fibroblasts are key players in cardiomyopathy progression, potentially worsening myocardial fibrosis and cardiac dysfunction. Targeting these specific fibroblast subpopulations offers new therapeutic avenues for treating cardiomyopathy.
Area of Science:
- Cardiology
- Molecular Biology
- Genomics
Background:
- Cardiomyopathy involves myocardial diseases leading to heart failure and arrhythmias.
- Myocardial fibrosis is a key pathological process in cardiomyopathy, driven by cardiac fibroblasts.
- Limited understanding exists regarding cardiac fibroblast plasticity and transcriptional regulation in cardiomyopathy.
Purpose of the Study:
- To conduct a comprehensive single-cell analysis of cardiac fibroblasts in cardiomyopathy.
- To explore differences in cellular plasticity and transcriptional regulatory networks among fibroblast subpopulations.
- To provide references for cardiomyopathy diagnosis, prognosis, and treatment.
Main Methods:
- Single-cell RNA sequencing and analysis of 179,927 cells.
- Clustering into 32 cell types, including 4 cardiac fibroblast subpopulations.
- Gene Ontology (GO) analysis, AUCell, Monocle, and SCENIC for transcriptional analysis.
Main Results:
- Identified 4 cardiac fibroblast subpopulations: C0 THBS4+, C1 LINC01133+, C2 FGF7+, and C3 AGT+ Fibroblasts.
- C3 AGT+ Fibroblasts are linked to immune response, protein transport, and myocardial function, correlating with disease progression.
- FOS identified as a key transcription factor in C3 AGT+ Fibroblasts, associated with proliferation and protein hydrolysis.
Conclusions:
- C3 AGT+ Fibroblasts show heightened sensitivity to adverse cardiomyopathy outcomes like fibrosis and impaired function.
- Differential activity of fibroblast subpopulations offers novel therapeutic targets for cardiomyopathy.
- Stemness genes EPAS1, MYC, and regulator FOS may modulate cardiac fibroblast biology in cardiomyopathy.
Background:
Cardiomyopathy encompasses a broad spectrum of diseases affecting myocardial tissue, characterized clinically by abnormalities in cardiac structure, heart failure, and/or arrhythmias. Clinically heterogeneous, major types include dilated cardiomyopathy (DCM), hypertrophic cardiomyopathy (HCM), restrictive cardiomyopathy (RM), ischemic cardiomyopathy (ICM), among which DCM is more prevalent, while ICM exhibits higher incidence and mortality rates. Myocardial injury during cardiomyopathy progression may lead to myocardial fibrosis. Failure to intervene early and inhibit the process of myocardial fibrosis may culminate in heart failure. Cardiac fibroblasts constitute crucial cellular components determining the extent and quality of myocardial fibrosis, with various subpopulations exerting diverse roles in cardiomyopathy progression. Despite this, understanding of the cellular plasticity and transcriptional regulatory networks of cardiac fibroblasts in cardiomyopathy remains limited. Therefore, in this study, we conducted comprehensive single-cell analysis of cardiac fibroblasts in cardiomyopathy to explore differences in cellular plasticity and transcriptional regulatory networks among fibroblast subpopulations, with the aim of providing as many useful references as possible for the diagnosis, prognosis, and treatment of cardiomyopathy.
Materials And Methods:
Cells with mitochondrial gene expression comprising >20% of total expressed genes were excluded. Differential expression genes (DEGs) and stemness genes within cardiac fibroblast subpopulations were subjected to Gene Ontology (GO) analysis of biological processes (BP) and AUCell analysis. Monocle software was employed to analyze the pseudo-temporal trajectory of cardiac fibroblasts in cardiomyopathy. Additionally, the Python package SCENIC was utilized to assess enrichment of transcription factors and activity of regulators within cardiac fibroblast subpopulations in cardiomyopathy.
Results:
Following batch effect correction, 179,927 cells were clustered into 32 clusters, designated as T_NK cells, endothelial cells, myeloid cells, fibroblasts, pericytes, SMCs, CMs, proliferating cells, EndoCs, and EPCs. Among them, 8148 fibroblasts were further subdivided into 4 subpopulations, namely C0 THBS4+ Fibroblasts, C1 LINC01133+ Fibroblasts, C2 FGF7+ Fibroblasts, and C3 AGT + Fibroblasts. Results from GO_BP and AUCell analyses suggest that C3 AGT + Fibroblasts may be associated with immune response activation, protein transport, and myocardial contractile function, correlating with disease progression in cardiomyopathy. Transcription factor enrichment analysis indicates that FOS is the most significant TF in C3 AGT + Fibroblasts, also associated with the M1 module, possibly implicated in protein hydrolysis, intracellular DNA replication, and cell proliferation. Moreover, correlation analysis of transcriptional regulatory activity between fibroblast subpopulations reveals a more pronounced heterogeneity within C3 AGT + Fibroblasts in cardiomyopathy.
Conclusion:
C3 AGT + Fibroblasts exhibit increased sensitivity towards adverse outcomes in cardiomyopathy, such as myocardial fibrosis and impaired cardiac contractile function, compared to other cardiac fibroblast subpopulations. The differential cellular plasticity and transcriptional regulatory activity between C3 AGT + Fibroblasts and other subgroups offer new perspectives for targeting fibroblast subpopulation activity to treat cardiomyopathy. Additionally, stemness genes EPAS1 and MYC, along with the regulator FOS, may play roles in modulating the biological processes of cardiac fibroblasts in cardiomyopathy.
