Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Article
  • Published:

Endothelial plasticity drives aberrant vascularization and impedes cardiac repair after myocardial infarction

Nature Cardiovascular Research volume 1, pages 372–388 (2022)Cite this article

Subjects

Abstract

Myocardial infarction (MI) is a leading cause of death worldwide, largely because efficient interventions to restore cardiac function after MI are currently lacking. Here, we characterize vascular aberrancies induced by MI and propose to target acquired endothelial cell (EC) changes to normalize vessels and promote cardiac repair after MI. Single-cell transcriptome analyses of MI-associated ECs indicates that ECs acquire mesenchymal gene signatures that result in phenotypic and functional changes and lead to vessel abnormalities. We identify a platelet-derived growth factor (PDGF)–nuclear factor κB (NF-κB)–hypoxia-inducible factor 1-α (HIF-1α) axis that induces Snail expression and mesenchymal phenotypes in ECs under hypoxia, altogether causing aberrant vascularization. EC-specific knockout of platelet-derived growth factor receptor beta (PDGFR-β), pharmacological PDGFR inhibition or nanoparticle-based targeted PDGFR-β small interfering RNA delivery in mice attenuates vascular abnormalities in the infarcted tissue and improves cardiac repair after MI. These findings illustrate a mechanism controlling aberrant neovascularization after ischemia and suggest that targeting PDGF/Snail-mediated endothelial plasticity may offer opportunities for normalizing vasculature and treating ischemic heart diseases.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Fig. 1: scRNA-seq analysis reveals dynamic endothelial plasticity in MI tissue, which is associated with vessel aberrancy.
Fig. 2: ECs acquire mesenchymal phenotypes under MI conditions.
Fig. 3: Linear RNA amplification and transcriptome analysis identifies PDGF as a critical regulator of endothelial plasticity and dysfunction under MI.
Fig. 4: PDGF regulates EC plasticity under hypoxia via NF-κB- and HIF1-α-dependent Snail expression.
Fig. 5: PDGF induces Snail binding to the VE-cadherin promoter, leading to VE-cadherin downregulated expression and mesenchymal phenotypes in ECs.
Fig. 6: PDGFR-β deletion in ECs normalizes the vasculature and improves cardiac function recovery after MI.
Fig. 7: Endothelial-specific deletion of PDGFR-β promotes tissue repair and cardiomyocyte function after MI.
Fig. 8: Pharmacological inhibition or nanoparticle-mediated targeting of PDGFR improves cardiac function recovery after MI.

Similar content being viewed by others

Data availability

The RNA-seq data have been deposited in the National Center for Biotechnology Information’s Gene Expression Omnibus under accession no. GSE163772, while the single-cell and single-nuclei RNA-seq analysis of mouse cardiac tissues with MI have been deposited in the same publicly available database under the accession nos. GSE163956 and GSE193290, respectively. The data supporting the findings of this study are available within the paper and its supplementary information. Source data are provided with this paper.

References

  1. Ware, J. A. & Simons, M. Angiogenesis in ischemic heart disease. Nat. Med. 3, 158–164 (1997).

    Article  CAS  PubMed  Google Scholar 

  2. Harada, K. et al. Vascular endothelial growth factor administration in chronic myocardial ischemia. Am. J. Physiol. 270, H1791–H1802 (1996).

    CAS  PubMed  Google Scholar 

  3. Banai, S. et al. Angiogenic-induced enhancement of collateral blood flow to ischemic myocardium by vascular endothelial growth factor in dogs. Circulation 89, 2183–2189 (1994).

    Article  CAS  PubMed  Google Scholar 

  4. Henry, T. D. et al. The VIVA trial: Vascular endothelial growth factor in Ischemia for Vascular Angiogenesis. Circulation 107, 1359–1365 (2003).

    Article  CAS  PubMed  Google Scholar 

  5. Stewart, D. J. et al. VEGF gene therapy fails to improve perfusion of ischemic myocardium in patients with advanced coronary disease: results of the NORTHERN trial. Mol. Ther. 17, 1109–1115 (2009).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  6. Rajagopalan, S. et al. Regional angiogenesis with vascular endothelial growth factor in peripheral arterial disease: a phase II randomized, double-blind, controlled study of adenoviral delivery of vascular endothelial growth factor 121 in patients with disabling intermittent claudication. Circulation 108, 1933–1938 (2003).

    Article  CAS  PubMed  Google Scholar 

  7. Potente, M., Gerhardt, H. & Carmeliet, P. Basic and therapeutic aspects of angiogenesis. Cell 146, 873–887 (2011).

    Article  CAS  PubMed  Google Scholar 

  8. Goel, S. et al. Normalization of the vasculature for treatment of cancer and other diseases. Physiol. Rev. 91, 1071–1121 (2011).

    Article  CAS  PubMed  Google Scholar 

  9. Kovacic, J. C., Mercader, N., Torres, M., Boehm, M. & Fuster, V. Epithelial-to-mesenchymal and endothelial-to-mesenchymal transition: from cardiovascular development to disease. Circulation 125, 1795–1808 (2012).

    Article  PubMed  PubMed Central  Google Scholar 

  10. Piera-Velazquez, S. & Jimenez, S. A. Endothelial to mesenchymal transition: role in physiology and in the pathogenesis of human diseases. Physiol. Rev. 99, 1281–1324 (2019).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  11. Zeisberg, E. M. et al. Endothelial-to-mesenchymal transition contributes to cardiac fibrosis. Nat. Med. 13, 952–961 (2007).

    Article  CAS  PubMed  Google Scholar 

  12. Maddaluno, L. et al. EndMT contributes to the onset and progression of cerebral cavernous malformations. Nature 498, 492–496 (2013).

    Article  CAS  PubMed  Google Scholar 

  13. Manavski, Y. et al. Clonal expansion of endothelial cells contributes to ischemia-induced neovascularization. Circ. Res. 122, 670–677 (2018).

    Article  CAS  PubMed  Google Scholar 

  14. Li, Z. et al. Single-cell transcriptome analyses reveal novel targets modulating cardiac neovascularization by resident endothelial cells following myocardial infarction. Eur. Heart J. 40, 2507–2520 (2019).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  15. Tombor, L. S. et al. Single cell sequencing reveals endothelial plasticity with transient mesenchymal activation after myocardial infarction. Nat. Commun. 12, 681 (2021).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  16. Huang, M. et al. c-Met-mediated endothelial plasticity drives aberrant vascularization and chemoresistance in glioblastoma. J. Clin. Invest. 126, 1801–1814 (2016).

    Article  PubMed  PubMed Central  Google Scholar 

  17. Fan, Y. Vascular detransformation for cancer therapy. Trends Cancer 5, 460–463 (2019).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  18. De Palma, M. et al. Tie2 identifies a hematopoietic lineage of proangiogenic monocytes required for tumor vessel formation and a mesenchymal population of pericyte progenitors. Cancer Cell 8, 211–226 (2005).

    Article  CAS  PubMed  Google Scholar 

  19. Andrae, J., Gallini, R. & Betsholtz, C. Role of platelet-derived growth factors in physiology and medicine. Genes Dev. 22, 1276–1312 (2008).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  20. Acloque, H., Adams, M. S., Fishwick, K., Bronner-Fraser, M. & Nieto, M. A. Epithelial-mesenchymal transitions: the importance of changing cell state in development and disease. J. Clin. Invest. 119, 1438–1449 (2009).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  21. Kalluri, R. & Weinberg, R. A. The basics of epithelial-mesenchymal transition. J. Clin. Invest. 119, 1420–1428 (2009).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  22. Lamouille, S., Xu, J. & Derynck, R. Molecular mechanisms of epithelial–mesenchymal transition. Nat. Rev. Mol. Cell Biol. 15, 178–196 (2014).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  23. Liu, T. et al. PDGF-mediated mesenchymal transformation renders endothelial resistance to anti-VEGF treatment in glioblastoma. Nat. Commun. 9, 3439 (2018).

    Article  PubMed  PubMed Central  Google Scholar 

  24. Khan, O. F. et al. Endothelial siRNA delivery in nonhuman primates using ionizable low-molecular weight polymeric nanoparticles. Sci. Adv. 4, eaar8409 (2018).

    Article  PubMed  PubMed Central  Google Scholar 

  25. Krohn-Grimberghe, M. et al. Nanoparticle-encapsulated siRNAs for gene silencing in the haematopoietic stem-cell niche. Nat. Biomed. Eng. 4, 1076–1089 (2020).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  26. Hoeben, A. et al. Vascular endothelial growth factor and angiogenesis. Pharmacol. Rev. 56, 549–580 (2004).

    Article  CAS  PubMed  Google Scholar 

  27. Forsythe, J. A. et al. Activation of vascular endothelial growth factor gene transcription by hypoxia-inducible factor 1. Mol. Cell. Biol. 16, 4604–4613 (1996).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  28. Oladipupo, S. et al. VEGF is essential for hypoxia-inducible factor-mediated neovascularization but dispensable for endothelial sprouting. Proc. Natl Acad. Sci. USA 108, 13264–13269 (2011).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  29. Nagy, J. A., Dvorak, A. M. & Dvorak, H. F. VEGF-A and the induction of pathological angiogenesis. Annu. Rev. Pathol. 2, 251–275 (2007).

    Article  CAS  PubMed  Google Scholar 

  30. Carmeliet, P. & Jain, R. K. Principles and mechanisms of vessel normalization for cancer and other angiogenic diseases. Nat. Rev. Drug Discov. 10, 417–427 (2011).

    Article  CAS  PubMed  Google Scholar 

  31. Potenta, S., Zeisberg, E. & Kalluri, R. The role of endothelial-to-mesenchymal transition in cancer progression. Br. J. Cancer 99, 1375–1379 (2008).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  32. Chen, P.-Y. et al. Endothelial-to-mesenchymal transition drives atherosclerosis progression. J. Clin. Invest. 125, 4514–4528 (2015).

    Article  PubMed  PubMed Central  Google Scholar 

  33. Dejana, E., Hirschi, K. K. & Simons, M. The molecular basis of endothelial cell plasticity. Nat. Commun. 8, 14361 (2017).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  34. Thiery, J. P. Epithelial–mesenchymal transitions in tumour progression. Nat. Rev. Cancer 2, 442–454 (2002).

    Article  CAS  PubMed  Google Scholar 

  35. Peinado, H., Olmeda, D. & Cano, A. Snail, Zeb and bHLH factors in tumour progression: an alliance against the epithelial phenotype? Nat. Rev. Cancer 7, 415–428 (2007).

    Article  CAS  PubMed  Google Scholar 

  36. Nieto, M. A. The snail superfamily of zinc-finger transcription factors. Nat. Rev. Mol. Cell Biol. 3, 155–166 (2002).

    Article  CAS  PubMed  Google Scholar 

  37. Bachelder, R. E., Yoon, S.-O., Franci, C., de Herreros, A. G. & Mercurio, A. M. Glycogen synthase kinase-3 is an endogenous inhibitor of Snail transcription: implications for the epithelial–mesenchymal transition. J. Cell Biol. 168, 29–33 (2005).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  38. Pires, B. R. B. et al. NF-kappaB is involved in the regulation of EMT genes in breast cancer cells. PLoS ONE 12, e0169622 (2017).

    Article  PubMed  PubMed Central  Google Scholar 

  39. Zhu, Y. et al. HIF-1α regulates EMT via the Snail and β-catenin pathways in paraquat poisoning-induced early pulmonary fibrosis. J. Cell. Mol. Med. 20, 688–697 (2016).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  40. Zhang, L. et al. Hypoxia induces epithelial-mesenchymal transition via activation of SNAI1 by hypoxia-inducible factor-1α in hepatocellular carcinoma. BMC Cancer 13, 108 (2013).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  41. Xu, X. et al. Snail is a direct target of hypoxia-inducible factor 1α (HIF1α) in hypoxia-induced endothelial to mesenchymal transition of human coronary endothelial cells. J. Biol. Chem. 290, 16653–16664 (2015).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  42. Kokudo, T. et al. Snail is required for TGFβ-induced endothelial-mesenchymal transition of embryonic stem cell-derived endothelial cells. J. Cell Sci. 121, 3317–3324 (2008).

    Article  CAS  PubMed  Google Scholar 

  43. Mahmoud, M. M. et al. Shear stress induces endothelial-to-mesenchymal transition via the transcription factor Snail. Sci. Rep. 7, 3375 (2017).

    Article  PubMed  PubMed Central  Google Scholar 

  44. Cheng, J.-C., Chang, H.-M. & Leung, P. C. K. Transforming growth factor-β1 inhibits trophoblast cell invasion by inducing Snail-mediated down-regulation of vascular endothelial-cadherin protein. J. Biol. Chem. 288, 33181–33192 (2013).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  45. Lopez, D., Niu, G., Huber, P. & Carter, W. B. Tumor-induced upregulation of Twist, Snail, and Slug represses the activity of the human VE-cadherin promoter. Arch. Biochem. Biophys. 482, 77–82 (2009).

    Article  CAS  PubMed  Google Scholar 

  46. Ishikawa, F. et al. Identification of angiogenic activity and the cloning and expression of platelet-derived endothelial cell growth factor. Nature 338, 557–562 (1989).

    Article  CAS  PubMed  Google Scholar 

  47. Benjamin, L. E., Hemo, I. & Keshet, E. A plasticity window for blood vessel remodelling is defined by pericyte coverage of the preformed endothelial network and is regulated by PDGF-B and VEGF. Development 125, 1591–1598 (1998).

    Article  CAS  PubMed  Google Scholar 

  48. Battegay, E. J., Rupp, J., Iruela-Arispe, L., Sage, E. H. & Pech, M. PDGF-BB modulates endothelial proliferation and angiogenesis in vitro via PDGF beta-receptors. J. Cell Biol. 125, 917–928 (1994).

    Article  CAS  PubMed  Google Scholar 

  49. Liu, C. et al. Platelet-derived growth factor blockade on cardiac remodeling following infarction. Mol. Cell. Biochem. 397, 295–304 (2014).

    Article  CAS  PubMed  Google Scholar 

  50. Zymek, P. et al. The role of platelet-derived growth factor signaling in healing myocardial infarcts. J. Am. Coll. Cardiol. 48, 2315–2323 (2006).

    Article  CAS  PubMed  Google Scholar 

  51. Wang, Y. et al. Ephrin-B2 controls VEGF-induced angiogenesis and lymphangiogenesis. Nature 465, 483–486 (2010).

    Article  CAS  PubMed  Google Scholar 

  52. Gong, Y., Zhao, Y., Li, Y., Fan, Y. & Hoover-Plow, J. Plasminogen regulates cardiac repair after myocardial infarction through its noncanonical function in stem cell homing to the infarcted heart. J. Am. Coll. Cardiol. 63, 2862–2872 (2014).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  53. Lang, R. M. et al. Recommendations for cardiac chamber quantification by echocardiography in adults: an update from the American Society of Echocardiography and the European Association of Cardiovascular Imaging. Eur. Heart J. Cardiovasc. Imaging 16, 233–270 (2015).

    Article  PubMed  Google Scholar 

  54. Qiu, X. et al. Single-cell mRNA quantification and differential analysis with Census. Nat. Methods 14, 309–315 (2017).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  55. Liu, T. et al. Circulating glioma cells exhibit stem cell-like properties. Cancer Res. 78, 6632–6642 (2018).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  56. Yang, B. et al. Single-cell phenotyping within transparent intact tissue through whole-body clearing. Cell 158, 945–958 (2014).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  57. Billingsley, M. M. et al. Ionizable lipid nanoparticle-mediated mRNA delivery for human CAR T cell engineering. Nano Lett. 20, 1578–1589 (2020).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

Download references

Acknowledgements

We thank J. Eberwine (Penn Pharmacology), J. Schug (Penn Next-Generation Sequencing Core), A. Stout (Penn CDB Microscope Core) and E. Blankemeyer (Penn Small Animal Imaging Core) for assistance with RNA linear amplification, scRNA-seq, light sheet fluorescence imaging and SPECT analysis, respectively. This work was supported in part by an American Heart Association (AHA) Innovative Project Award no. IPA34170252 (to Y.F.), National Institutes of Health (NIH) grant nos. R01HL155198 (to Y.F. and Y.G.) and R01NS094533, R01NS106108 and R01CA241501 (to Y.F.), American Association for Cancer Research (AACR) Judah Folkman Award (to Y.F.), AHA Scientist Development grant no. SDG9050018 and grant-in-Aid no. GRNT3365002 (to Y.G.) and AHA Predoctoral Fellowship (to D.Z.). L.P. is supported by the Office of the Assistant Secretary of Defense for Health Affairs through the Peer Reviewed Medical Research Program (nos. W81XWH2010042 and W81XWH2010089). M.J.M. acknowledges support from a Burroughs Wellcome Fund Career Award at the Scientific Interface, an NIH Director’s New Innovator Award (no. DP2 TR002776), NIH grant nos. R01CA241661, R37CA244911 and R01DK123049, an Abramson Cancer Center-School of Engineering and Applied Sciences Discovery Grant (no. P30CA016520) and an AACR-Bayer Innovation and Discovery Grant (no. 18-80-44-MITC).

Author information

Author notes

  1. Maohuan Lin

    Present address: Department of Cardiology, Sun Yat-sen Memorial Hospital, Sun Yat-sen University, Guangzhou, China

  2. These authors contributed equally: Menggui Huang, Fan Yang, Duo Zhang, Maohuan Lin.

  3. These authors jointly supervised this work: Yi Fan, Yanqing Gong.

Authors and Affiliations

  1. Department of Radiation Oncology, University of Pennsylvania Perelman School of Medicine, Philadelphia, PA, USA

    Menggui Huang, Fan Yang, Duo Zhang, Hao Duan & Yi Fan

  2. Division of Human Genetics and Translational Medicine, Department of Medicine, University of Pennsylvania Perelman School of Medicine, Philadelphia, PA, USA

    Maohuan Lin, Daniel J. Rader & Yanqing Gong

  3. Department of Bioengineering, University of Pennsylvania School of Engineering and Applied Science, Philadelphia, PA, USA

    Rakan El-Mayta & Michael J. Mitchell

  4. Department of Obstetrics & Gynecology, University of Pennsylvania Perelman School of Medicine, Philadelphia, PA, USA

    Lin Zhang

  5. Department of Orthopaedic Surgery, University of Pennsylvania Perelman School of Medicine, Philadelphia, PA, USA

    Ling Qin

  6. Cardiovascular Institute, University of Pennsylvania Perelman School of Medicine, Philadelphia, PA, USA

    Swapnil V. Shewale, Liming Pei, Michael J. Mitchell, Daniel J. Rader, Yi Fan & Yanqing Gong

  7. Center for Mitochondrial and Epigenomic Medicine, Department of Pathology and Laboratory Medicine, Children’s Hospital of Philadelphia, Philadelphia, PA, USA

    Liming Pei

  8. Abramson Cancer Center, University of Pennsylvania Perelman School of Medicine, Philadelphia, PA, USA

    Michael J. Mitchell & Yi Fan

  9. Institute for Immunology, University of Pennsylvania Perelman School of Medicine, Philadelphia, PA, USA

    Michael J. Mitchell & Yi Fan

  10. Institute for Regenerative Medicine, University of Pennsylvania Perelman School of Medicine, Philadelphia, PA, USA

    Michael J. Mitchell

Authors
  1. Menggui Huang
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Fan Yang
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Duo Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Maohuan Lin
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Hao Duan
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Rakan El-Mayta
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Lin Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Ling Qin
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Swapnil V. Shewale
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Liming Pei
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. Michael J. Mitchell
    View author publications

    You can also search for this author in PubMed Google Scholar

  12. Daniel J. Rader
    View author publications

    You can also search for this author in PubMed Google Scholar

  13. Yi Fan
    View author publications

    You can also search for this author in PubMed Google Scholar

  14. Yanqing Gong
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

M.H., F.Y., D.Z. and M.L. designed, performed and analyzed the experiments. D.Z., L.Z. and L.Q. contributed to the scRNA-seq studies. M.L. and S.V.S. contributed to the echocardiogram analysis. H.D. helped tissue histology analysis. R.E. and M.J.M. contributed to nanoparticle production. L.P. helped with the single-nucleus analysis. D.J.R. contributed to the experimental design and discussion. Y.G. and M.L. performed the MI surgery. Y.F. and Y.G. cosupervised the experiments and wrote the manuscript. All authors commented on the manuscript.

Corresponding authors

Correspondence to Yi Fan or Yanqing Gong.

Ethics declarations

Competing interests

The authors declare no competing interests.

Peer review

Peer review information

Nature Cardiovascular Research thanks the anonymous reviewers for their contribution to the peer review of this work.

Additional information

Publisher’s note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Integrated supplementary information

Extended Data Fig. 1 Aberrant vascularization characterizes infarcted cardiac tissue.

Mice were subjected to MI surgery or sham operation. Hearts were excised 3 weeks after surgery. Tissue sections were immunostained with (a) anti-CD31 antibody, (b) anti-NG2 and anti-CD31 antibodies, or (c) anti-collagen IV and anti-CD31 antibodies. a, Representative images in normal tissue and infarct zone are shown (n = 3 mice). Bars indicate 100 µm. b,c, Left, representative images in normal tissue and infarct zone are shown. Right, quantified results (mean ± SEM, n = 3 mice). Statistical analysis by unpaired two-tailed Students’ t test. Bars indicate 100 µm.

Source Data

Extended Data Fig. 2 Cardiomyocyte-conditioned medium induces FSP-1 and α-SMA expression in ECs.

Mouse cardiac-conditioned medium (CCM) were harvested from primary mouse cardiomyocytes cultured under hypoxia (1% O2). Mouse cardiac microvascular ECs were treated with CCM under normoxia or hypoxia for 2 days. Cell lysates were immunoblotted. These experiments were repeated independently twice with similar results.

Source Data

Extended Data Fig. 3 Expression of mesenchymal genes in MI-associated ECs.

MI was induced in wild-type mice. Three weeks after MI induction, ECs were isolated from normal LV and MI tissues. RNA was extracted and subjected to RT-PCR analysis (mean ± SEM, n = 5-7 mice, specific n indicated in the graphs). Statistical analysis by unpaired two-tailed Students’ t test. Expression of (a) S100A4 (FSP-1) and Acta2 (α-SMA), and (b) Snai1 (Snail), Snai2 (Slug), and Twist1 was normalized with GAPDH expression levels.

Source Data

Extended Data Fig. 4 PDGF-AB induces expression of mesenchymal proteins in ECs.

Human cardiac microvascular ECs were treated with 100 ng/ml SCF, PDGF-AB, or HGF for 2 days under hypoxia. Cell lysates were immunoblotted. These experiments were repeated independently twice with similar results.

Source Data

Extended Data Fig. 5 PDGFR-β knockout inhibits PDGF-AB-induced phosphorylation of Erk1 and Akt1 in ECs and does not affect vascular density in normal hearts.

a, Aortic ECs were isolated from tamoxifen-treated Cdh5-CreERT2;Pdgfrbfl/fl (PDGFR-β-ΔEC) and Pdgfrbfl/fl (control) mice. Cells were treated with 100 ng/ml PDGF-AB for 10 min, followed by immunoblot analysis. These experiments were repeated independently twice with similar results. b,Left ventricles were collected from PDGFR-β-ΔEC and control mice. Cardiac sections were stained with anti-CD31 antibody and analyzed by immunofluorescence. Left, representative images are shown. Scale bar: 200 μm. Right, quantified results (mean ± SEM, n = 6 mice). Statistical analysis by unpaired two-tailed Students’ t test.

Source Data

Extended Data Fig. 6 PDGFR-β knockout in ECs does not increase vascular density in infarct zone and border zone.

MI was induced in control and PDGFR-β-ΔEC mice. (a,b) 4 or (c) 8 weeks after MI induction, cardiac tissues were collected. Sections were stained with anti-collagen I and anti-CD31 antibodies, followed by immunofluorescence analysis. a, Representative images are shown (n = 6 mice). Scale bar: 100 μm. b,c, Vascular density was quantified (mean ± SEM). b, n = 3-4 mice, specific n indicated in the graphs. c, n = 6 mice. Statistical analysis by two-way ANOVA Fisher’s test.

Source Data

Extended Data Fig. 7 PDGFR-β knockout reduces vascular density and inhibits Ki67, Snail and FSP-1 expression in MI- associated ECs.

MI was induced in WT and PDGFR-β-ΔEC mice. Hearts were excised 3 weeks after surgery. MI tissue sections were immunostained with (a-c) anti-Ki67 and anti-CD31, (d) anti-vWF and anti-Snail, or (e) anti-CD31 and anti-FSP-1 antibodies. a, Representative immunofluorescence images in infarct zone are shown (n = 3 mice). Scale bar: 100 μm. b,c, quantified results (mean ± SEM, n = 3 mice). b, Density of CD31+ cells. AU, arbitrary unit. c, Ki67 expression in CD31+ cells. Statistical analysis by unpaired two-tailed Students’ t test. d,e, Representative immunofluorescence images in infarct zone are shown (n = 5 mice). Scale bar: 100 μm.

Source Data

Extended Data Fig. 8 PDGFR-β knockout reduces Snail expression in MI-associated ECs.

MI was induced in WT and PDGFR-β-ΔEC mice. (a,b) 14 and (c,d) 28 days after MI induction, MI tissue sections were collected and subjected to single-nuclei RNA sequencing analysis (total = 4 mice). Uniform manifold approximation and projection (UMAP) analysis of transcriptome gene signature in ECs. (a,c) UMAP analysis of ECs. (b,d) Expression of Snail, Slug, and Ki67 in ECs.

Extended Data Fig. 9 PDGFR-β knockout in ECs alters expression of metabolism-associated genes in myocytes.

MI was induced in WT and PDGFR-β-ΔEC mice. 14 and 28 days after MI induction, MI tissue sections were collected and subjected to single-nuclei RNA sequencing analysis (total = 4 mice). Expression of metabolism-associated genes were analyzed. a, Heatmap of mapped genes. b, Global changes in gene expression at days 14 and 28 after MI induction.

Extended Data Fig. 10 PDGFR inhibition improves cardiac function recovery and promotes tissue repair after MI.

MI was induced in mice, followed by administration with saline or 15 mg/kg crenolanib. a,b, Cardiac function was analyzed by echocardiogram (n = 11 mice). Representative images are shown. a, Long-axis echocardiogram analysis. Scale bar: 2 mm. b, M mode echocardiogram analysis. Scale bar: 2 mm. Arrows indicate myocardial anterior walls. c,d, Heart tissues were harvested 3 weeks after MI induction. Cardiac sections were stained with Masson’s trichrome stain. c, Representative images are shown (n = 4 mice). Scale bar: 500 μm. Arrows indicate epicardium. d, Quantified cardiomyocyte area (mean ± SEM, n = 4-5 mice). Statistical analysis by unpaired two-tailed Students’ t test. e, Heart tissues were harvested 3 weeks after MI induction. Cardiac sections were stained with an anti-CD31 antibody and analyzed by immunofluorescence imaging. Vascular density was quantified (mean ± SEM, n = 5 mice). Statistical analysis by two-way ANOVA.

Source Data

Supplementary information

Reporting Summary

Supplementary Video 1.

MI was induced in control and PDGFR-β-ΔEC mice. Mice underwent echocardiography analysis at days 14 and 28 after MI.

Source data

Source Data Fig. 1

Statistical source data.

Source Data Fig. 2

Statistical source data.

Source Data Fig. 2

Unprocessed western blots.

Source Data Fig. 3

Statistical source data.

Source Data Fig. 3

Unprocessed western blots.

Source Data Fig. 4

Statistical source data.

Source Data Fig. 4

Unprocessed western blots.

Source Data Fig. 5

Statistical source data.

Source Data Fig. 5

Unprocessed western blots.

Source Data Fig. 6

Statistical source data.

Source Data Fig. 6

Unprocessed western blots.

Source Data Fig. 7

Statistical source data.

Source Data Fig. 8

Statistical source data.

Source Data Fig. 8

Unprocessed western blots.

Source Data Extended Data Fig. 1

Statistical source data.

Source Data Extended Data Fig. 2

Unprocessed western blots.

Source Data Extended Data Fig. 3

Statistical source data.

Source Data Extended Data Fig. 4

Unprocessed western blots.

Source Data Extended Data Fig. 5

Statistical source data.

Source Data Extended Data Fig. 5

Unprocessed western blots.

Source Data Extended Data Fig. 6

Statistical source data.

Source Data Extended Data Fig. 7

Statistical source data.

Source Data Extended Data Fig. 10

Statistical source data.

Rights and permissions

Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Huang, M., Yang, F., Zhang, D. et al. Endothelial plasticity drives aberrant vascularization and impedes cardiac repair after myocardial infarction. Nat Cardiovasc Res 1, 372–388 (2022). https://doi.org/10.1038/s44161-022-00047-3

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/s44161-022-00047-3

This article is cited by

Search

Advanced search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing