Cardiac fibroblasts regulate myocardium and coronary vasculature development in the murine heart via the collagen signaling pathway

The fibroblast (FB), cardiomyocyte (CM), and vascular endothelial cell (Vas_EC) are the three major cell types in the heart, yet their relationships during development are largely unexplored. To address this gap, we employed RNA staining of the FB marker gene Col1a1 together with the CM marker gene...

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Main Authors: Yiting Deng, Yuanhang He, Juan Xu, Haoting He, Manling Zhang, Guang Li
Format: Article
Language:English
Published: eLife Sciences Publications Ltd 2025-07-01
Series:eLife
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Online Access:https://elifesciences.org/articles/102305
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author Yiting Deng
Yuanhang He
Juan Xu
Haoting He
Manling Zhang
Guang Li
author_facet Yiting Deng
Yuanhang He
Juan Xu
Haoting He
Manling Zhang
Guang Li
author_sort Yiting Deng
collection DOAJ
description The fibroblast (FB), cardiomyocyte (CM), and vascular endothelial cell (Vas_EC) are the three major cell types in the heart, yet their relationships during development are largely unexplored. To address this gap, we employed RNA staining of the FB marker gene Col1a1 together with the CM marker gene Actn2 and the Vas_EC marker gene Cdh5 at various stages of mouse heart development. This approach enabled us to discern the anatomical pattern of cardiac FBs and identify approximately one EC and four CMs directly interacting with each FB. Molecularly, through the analysis of single-cell mRNA sequencing (scRNA-seq) data, we unveiled collagen as the top signaling molecule derived from FBs influencing CM and Vas_EC development. Subsequently, we used a Pdgfra-CreER controlled diphtheria toxin A (DTA) system to ablate the FBs at different stages. We found that the ablation of FBs disrupted myocardium and vasculature development and led to embryonic heart defects. Using scRNA-seq, we further profiled the ablated hearts and identified molecular defects in their ventricular CMs and Vas_ECs compared to control hearts. Moreover, we identified a reduction of collagen in the ablated hearts and predicted collagen as the major signaling pathway regulating the differentially expressed genes in the ablated ventricular CMs. Finally, we performed both short-term and long-term FB ablation at the neonatal stage. We found that short-term ablation caused a reduction in collagen and Vas_EC density, while long-term ablation may induce compensatory collagen expression without causing heart function reduction. In summary, our study has identified the function of FBs in regulating myocardium and vasculature development in the mouse heart and implicated an important role for the collagen pathway in this process.
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spelling doaj-art-7dc7da9debeb4b19bd0dd6b7d7ebef622025-08-20T02:38:06ZengeLife Sciences Publications LtdeLife2050-084X2025-07-011310.7554/eLife.102305Cardiac fibroblasts regulate myocardium and coronary vasculature development in the murine heart via the collagen signaling pathwayYiting Deng0Yuanhang He1Juan Xu2https://orcid.org/0009-0004-3184-8730Haoting He3Manling Zhang4Guang Li5https://orcid.org/0000-0002-8546-2364Department of Cell Biology, University of Pittsburgh School of Medicine, Pittsburgh, United StatesDepartment of Cell Biology, University of Pittsburgh School of Medicine, Pittsburgh, United States; Tsinghua University, Tsinghua medicine, School of Medicine, Beijing, ChinaDepartment of Cell Biology, University of Pittsburgh School of Medicine, Pittsburgh, United StatesDepartment of Cell Biology, University of Pittsburgh School of Medicine, Pittsburgh, United StatesVascular Medicine Institute, Department of Medicine, University of Pittsburgh, Pittsburgh, United StatesDepartment of Cell Biology, University of Pittsburgh School of Medicine, Pittsburgh, United StatesThe fibroblast (FB), cardiomyocyte (CM), and vascular endothelial cell (Vas_EC) are the three major cell types in the heart, yet their relationships during development are largely unexplored. To address this gap, we employed RNA staining of the FB marker gene Col1a1 together with the CM marker gene Actn2 and the Vas_EC marker gene Cdh5 at various stages of mouse heart development. This approach enabled us to discern the anatomical pattern of cardiac FBs and identify approximately one EC and four CMs directly interacting with each FB. Molecularly, through the analysis of single-cell mRNA sequencing (scRNA-seq) data, we unveiled collagen as the top signaling molecule derived from FBs influencing CM and Vas_EC development. Subsequently, we used a Pdgfra-CreER controlled diphtheria toxin A (DTA) system to ablate the FBs at different stages. We found that the ablation of FBs disrupted myocardium and vasculature development and led to embryonic heart defects. Using scRNA-seq, we further profiled the ablated hearts and identified molecular defects in their ventricular CMs and Vas_ECs compared to control hearts. Moreover, we identified a reduction of collagen in the ablated hearts and predicted collagen as the major signaling pathway regulating the differentially expressed genes in the ablated ventricular CMs. Finally, we performed both short-term and long-term FB ablation at the neonatal stage. We found that short-term ablation caused a reduction in collagen and Vas_EC density, while long-term ablation may induce compensatory collagen expression without causing heart function reduction. In summary, our study has identified the function of FBs in regulating myocardium and vasculature development in the mouse heart and implicated an important role for the collagen pathway in this process.https://elifesciences.org/articles/102305fibroblastextracellular matrixablationheartsingle cellmyocardium
spellingShingle Yiting Deng
Yuanhang He
Juan Xu
Haoting He
Manling Zhang
Guang Li
Cardiac fibroblasts regulate myocardium and coronary vasculature development in the murine heart via the collagen signaling pathway
eLife
fibroblast
extracellular matrix
ablation
heart
single cell
myocardium
title Cardiac fibroblasts regulate myocardium and coronary vasculature development in the murine heart via the collagen signaling pathway
title_full Cardiac fibroblasts regulate myocardium and coronary vasculature development in the murine heart via the collagen signaling pathway
title_fullStr Cardiac fibroblasts regulate myocardium and coronary vasculature development in the murine heart via the collagen signaling pathway
title_full_unstemmed Cardiac fibroblasts regulate myocardium and coronary vasculature development in the murine heart via the collagen signaling pathway
title_short Cardiac fibroblasts regulate myocardium and coronary vasculature development in the murine heart via the collagen signaling pathway
title_sort cardiac fibroblasts regulate myocardium and coronary vasculature development in the murine heart via the collagen signaling pathway
topic fibroblast
extracellular matrix
ablation
heart
single cell
myocardium
url https://elifesciences.org/articles/102305
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