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An iPSC-derived vascular model of Marfan syndrome identifies key mediators of smooth muscle cell death

Nature Genetics volume 49, pages 97109 (2017) | Download Citation

Abstract

Marfan syndrome (MFS) is a heritable connective tissue disorder caused by mutations in FBN1, which encodes the extracellular matrix protein fibrillin-1. To investigate the pathogenesis of aortic aneurysms in MFS, we generated a vascular model derived from human induced pluripotent stem cells (MFS-hiPSCs). Our MFS-hiPSC-derived smooth muscle cells (SMCs) recapitulated the pathology seen in Marfan aortas, including defects in fibrillin-1 accumulation, extracellular matrix degradation, transforming growth factor-β (TGF-β) signaling, contraction and apoptosis; abnormalities were corrected by CRISPR-based editing of the FBN1 mutation. TGF-β inhibition rescued abnormalities in fibrillin-1 accumulation and matrix metalloproteinase expression. However, only the noncanonical p38 pathway regulated SMC apoptosis, a pathological mechanism also governed by Krüppel-like factor 4 (KLF4). This model has enabled us to dissect the molecular mechanisms of MFS, identify novel targets for treatment (such as p38 and KLF4) and provided an innovative human platform for the testing of new drugs.

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Acknowledgements

The authors thank L. Vallier, the hiPSC core facility at the Anne McLaren Laboratory and I. Geti for help in generating the MFS hiPSC lines. We thank N. Figg for sectioning and staining of the teratomas, J. Sterling for help with skin biopsies, J. Skepper at the Cambridge Advance Imaging Centre at the University of Cambridge for the transmission electron microscopy images and C. Verstreken at the Cambridge Stem Cell Institute for providing the static stretching membranes. We also thank Z. Mallat for useful comments on the manuscript. This work was supported by Evelyn Trust, the NIHR Cambridge Biomedical Research Centre and the British Heart Foundation (FS/13/29/30024 (S.S.), RM/l3/3/30159 (S.S. and M.M.) and FS/11/77/29327 (W.B.)).

Author information

Affiliations

  1. Stroke Research Group, Department of Clinical Neurosciences, Cambridge Biomedical Campus, University of Cambridge, Cambridge, UK.

    • Alessandra Granata
  2. Anne McLaren Laboratory, Wellcome Trust-MRC Cambridge Stem Cell Institute, University of Cambridge, Cambridge, UK.

    • Felipe Serrano
    • , William George Bernard
    • , Madeline McNamara
    • , Priya Sastry
    •  & Sanjay Sinha
  3. Division of Cardiovascular Medicine, University of Cambridge, Cambridge, UK.

    • Felipe Serrano
    • , William George Bernard
    • , Madeline McNamara
    • , Lucinda Low
    • , Priya Sastry
    •  & Sanjay Sinha
  4. British Heart Foundation Oxbridge Centre for Regenerative Medicine, University of Cambridge, Cambridge, UK.

    • Madeline McNamara
    •  & Sanjay Sinha
  5. Papworth Hospitals NHS Foundation Trust, Cambridge, UK.

    • Priya Sastry

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Contributions

A.G., conception, design, acquisition, analysis and interpretation of iPSC-SMC data and to the drafting of the article; F.S., help with conception and design and the analysis of the CRISPR–Cas9 iPSC clones and NC protocol; W.G.B., help with to the neural crest protocol for deriving NC-SMCs; M.M., qPCR analysis of fibroblasts and iPSC-derived SMC lines and the stretch studies; L.L., teratoma assay; P.S., providing human aortic tissue; S.S., conceptual design of the experiments, help with obtaining funding and supervision of all studies. All authors contributed to revision of the article.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Sanjay Sinha.

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    Supplementary Figures 1–15, Supplementary Tables 1–5 and Supplementary Note

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DOI

https://doi.org/10.1038/ng.3723

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