Original Article

Fibroblasts in an endocardial fibroelastosis disease model mainly originate from mesenchymal derivatives of epicardium

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Abstract

Endocardial fibroelastosis (EFE) refers to the thickening of the ventricular endocardium as a result of de novo deposition of subendocardial fibrous tissue layers during neonatal heart development. The origin of EFE fibroblasts is proposed to be postnatal endocardial cells that undergo an aberrant endothelial-to-mesenchymal transition (EndMT). Genetic lineage tracing of endocardial cells with the inducible endocardial Cre line Npr3-CreER and the endothelial cell tracing line Cdh5-CreER on an EFE-like model did not reveal any contribution of neonatal endocardial cells to fibroblasts in the EFE-like tissues. Instead, lineage tracing of embryonic epicardium by Wt1-CreER suggested that epicardium-derived mesenchymal cells (MCs) served as the major source of EFE fibroblasts. By labeling MCs using Sox9-CreER, we confirmed that MCs of the embryonic heart expand and contribute to the majority of neonatal EFE fibroblasts. During this pathological process, TGFβ signaling, the key mediator of fibroblasts activation, was highly upregulated in the EFE-like tissues. Targeting TGFβ signaling by administration of its antagonist bone morphogenetic protein 7 effectively reduced fibroblast accumulation and tissue fibrosis in the EFE-like model. Our study provides genetic evidence that excessive fibroblasts in the EFE-like tissues mainly originate from the epicardium-derived MCs through epicardial to mesenchymal transition (EpiMT). These EpiMT-derived fibroblasts within the EFE-like tissues could serve as a potential therapeutic target.

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Acknowledgements

We thank Baojin Wu, Guoyuan Chen, Zhonghui Weng and Aimin Huang for the animal husbandry; and Wei Bian for his technical help. We thank Shanghai Biomodel Organism Science & Technology Development Co., Ltd for mouse generation. We thank Ralf Adams at Max Plank Institute for providing the Cdh5-CreER mouse line and Hongkui Zeng for reporter lines. We also thank other members of our laboratory for insightful discussion and technical help throughout this study. This work was supported by Strategic Priority Research Program of the Chinese Academy of Sciences (CAS, XDB19000000), The National key Research & Development Program of China (2017YFC1001300 and 2016YFC1300600), National Natural Science Foundation of China (91639302, 31625019, 31571503, 31501172, 31601168), Youth Innovation Promotion Association of CAS (2015218), Key Project of Frontier Sciences of CAS (QYZDB-SSW-SMC003), International Cooperation Fund of CAS, National Program for Support of Top-notch Young Professionals, Shanghai Science and Technology Commission (14JC1407300, 17ZR1449600, 17ZR1449800), Shanghai Yangfan Project (15YF1414000, 16YF1413400) and Rising-Star Program (15QA1404300), China Postdoctoral Science Foundation (2015M581669, 2016T90387, 2016LH0042), President Fund of Shanghai Institutes for Biological Sciences (SIBS), Astrazeneca, Boehringer Ingelheim, Sanofi-SIBS Fellowship and Research Grants Council of Hong Kong (24110515, 14111916).

Author information

Author notes

    • Hui Zhang
    • , Xiuzhen Huang
    • , Kuo Liu
    •  & Juan Tang

    These four authors contributed equally to this work.

Affiliations

  1. The State Key Laboratory of Cell Biology, CAS Center for Excellence in Molecular Cell Science, Shanghai Institute of Biochemistry and Cell Biology, Chinese Academy of Sciences, University of Chinese Academy of Sciences, Shanghai 200031, China;

    • Hui Zhang
    • , Xiuzhen Huang
    • , Kuo Liu
    • , Juan Tang
    • , Lingjuan He
    • , Wenjuan Pu
    • , Qiaozhen Liu
    • , Yan Li
    • , Xueying Tian
    • , Yue Wang
    • , Libo Zhang
    • , Hongyan Wang
    • , Ronggui Hu
    •  & Bin Zhou
  2. Key Laboratory of Nutrition and Metabolism, Institute for Nutritional Sciences, Shanghai Institutes for Biological Sciences, Graduate School of the Chinese Academy of Sciences, Chinese Academy of Sciences, Shanghai 200031, China;

    • Hui Zhang
    • , Xiuzhen Huang
    • , Kuo Liu
    • , Juan Tang
    • , Lingjuan He
    • , Wenjuan Pu
    • , Qiaozhen Liu
    • , Yan Li
    • , Xueying Tian
    • , Yue Wang
    • , Libo Zhang
    • , Ying Yu
    •  & Bin Zhou
  3. School of Life Science and Technology, ShanghaiTech University, Shanghai 201210, China;

    • Hui Zhang
    • , Kuo Liu
    •  & Bin Zhou
  4. National Institute of Biological Sciences, Beijing 102206, China;

    • Fengchao Wang
    •  & Ting Chen
  5. Cardiovascular and Metabolic Diseases, Innovative Medicines and Early Clinical Development Biotech Unit, AstraZeneca, Mölndal 43183, Sweden;

    • Qing-Dong Wang
  6. Department of Cardiovascular Medicine, Southern Medical University Affiliated Fengxian Hospital, Shanghai 201499, China;

    • Zengyong Qiao
  7. Department of Cardiology, the First Affiliated Hospital, School of Medicine, Zhejiang University, 79 Qingchun Road, Hangzhou, Zhejiang 310003, China;

    • Li Zhang
  8. Department of Chemical Pathology, Li Ka Shing Institute of Health Sciences, The Chinese University of Hong Kong, Prince of Wales Hospital, Shatin, Hong Kong SAR 999077, China;

    • Kathy O Lui
  9. Key Laboratory of Regenerative Medicine of Ministry of Education, Institute of Aging and Regenerative Medicine, Jinan University, Guangzhou, Guangdong 510632, China

    • Bin Zhou

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Corresponding authors

Correspondence to Hui Zhang or Bin Zhou.

Supplementary information

PDF files

  1. 1.

    Supplementary information, Figure S1

    Endocardial fibroelastosis (EFE) tissue lining the left ventricle after heterotopic heart transplantation.

  2. 2.

    Supplementary information, Figure S2

    Npr3-CreER labeled endocardial or epicardial cells do not contribute to fibroblasts in native hearts.

  3. 3.

    Supplementary information, Figure S3

    Sox9 is expressed in cushion mesenchymal cells and epicardiai cells at E10.5 - E11.5.

  4. 4.

    Supplementary information, Figure S4

    Sox9-CreER labels embryonic cardiac mesenchymal cells and epicardial cells, but not endothelial cells.

  5. 5.

    Supplementary information, Figure S5

    Sox9-CreER does not label hematopoietic cells.

  6. 6.

    Supplementary information, Figure S6

    Embryonic endocardial cells contribute to fibroblasts in the native hearts.

  7. 7.

    Supplementary information, Figure S7

    FSP1 is expressed in leucocytes.

(Supplementary information is linked to the online version of the paper on the Cell Research website.)