Shear stimulation of FOXC1 and FOXC2 differentially regulates cytoskeletal activity during lymphatic valve maturation

  1. Pieter R Norden
  2. Amélie Sabine
  3. Ying Wang
  4. Cansaran Saygili Demir
  5. Ting Liu
  6. Tatiana V Petrova
  7. Tsutomu Kume  Is a corresponding author
  1. Northwestern University, United States
  2. University of Lausanne, Switzerland
  3. Mayo Clinic, United States

Abstract

Mutations in the transcription factor FOXC2 are predominately associated with lymphedema. Herein, we demonstrate a key role for related factor FOXC1, in addition to FOXC2, in regulating cytoskeletal activity in lymphatic valves. FOXC1 is induced by laminar, but not oscillatory, shear and inducible, endothelial-specific deletion impaired postnatal lymphatic valve maturation in mice. However, deletion of Foxc2 induced valve degeneration, which is exacerbated in Foxc1; Foxc2 mutants. FOXC1 knockdown (KD) in human lymphatic endothelial cells increased focal adhesions and actin stress fibers whereas FOXC2-KD increased focal adherens and disrupted cell junctions, mediated by increased ROCK activation. ROCK inhibition rescued cytoskeletal or junctional integrity changes induced by inactivation of FOXC1 and FOXC2 in vitro and vivo respectively, but only ameliorated valve degeneration in Foxc2 mutants. These results identify both FOXC1 and FOXC2 as mediators of mechanotransduction in the postnatal lymphatic vasculature and posit cytoskeletal signaling as a therapeutic target in lymphatic pathologies.

Data availability

All data generated or analysed during this study are included in the manuscript and supporting files.

The following previously published data sets were used

Article and author information

Author details

  1. Pieter R Norden

    Medicine, Northwestern University, Chicago, United States
    Competing interests
    The authors declare that no competing interests exist.
  2. Amélie Sabine

    Oncology, University of Lausanne, Epalinge, Switzerland
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-7512-6703
  3. Ying Wang

    Biochemistry and Molecular Biology, Mayo Clinic, Jacksonville, United States
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-7852-386X
  4. Cansaran Saygili Demir

    Oncology, University of Lausanne, Epalinges, Switzerland
    Competing interests
    The authors declare that no competing interests exist.
  5. Ting Liu

    Medicine, Northwestern University, Chicago, United States
    Competing interests
    The authors declare that no competing interests exist.
  6. Tatiana V Petrova

    Oncology, University of Lausanne, Epalinges, Switzerland
    Competing interests
    The authors declare that no competing interests exist.
  7. Tsutomu Kume

    Medicine, Northwestern University, Chicago, United States
    For correspondence
    t-kume@northwestern.edu
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-6005-5316

Funding

National Institutes of Health (R01HL126920,R01HL144129)

  • Tsutomu Kume

National Institutes of Health (5T32HL094293)

  • Pieter R Norden

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.

Ethics

Animal experimentation: All procedures and animal studies were approved by Northwestern University's institutional animal care and use committee (IACUC) (Protocol: IS00008794) or by the Animal Ethics Committee of Vaud, Switzerland.

Reviewing Editor

  1. Natasha L Harvey, University of South Australia, Australia

Publication history

  1. Received: November 21, 2019
  2. Accepted: June 6, 2020
  3. Accepted Manuscript published: June 8, 2020 (version 1)
  4. Version of Record published: June 18, 2020 (version 2)

Copyright

© 2020, Norden et al.

This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.

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  1. Pieter R Norden
  2. Amélie Sabine
  3. Ying Wang
  4. Cansaran Saygili Demir
  5. Ting Liu
  6. Tatiana V Petrova
  7. Tsutomu Kume
(2020)
Shear stimulation of FOXC1 and FOXC2 differentially regulates cytoskeletal activity during lymphatic valve maturation
eLife 9:e53814.
https://doi.org/10.7554/eLife.53814
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