Discovery of novel determinants of endothelial lineage using chimeric heterokaryons

  1. Wing Tak Wong
  2. Gianfranco Matrone
  3. XiaoYu Tian
  4. Simion Alin Tomoiaga
  5. Kin Fai Au
  6. Shu Meng
  7. Sayumi Yamazoe
  8. Daniel Sieveking
  9. Kaifu Chen
  10. David M Burns
  11. James K Chen
  12. Helen M Blau
  13. John P Cooke  Is a corresponding author
  1. Houston Methodist Research Institute, United States
  2. University of Iowa, United States
  3. Stanford University School of Medicine, United States

Abstract

We wish to identify determinants of endothelial lineage. Murine embryonic stem cells (mESC) were fused with human endothelial cells in stable, non-dividing, heterokaryons. Using RNA-seq it is possible to discriminate between human and mouse transcripts in these chimeric heterokaryons. We observed a temporal pattern of gene expression in the ESCs of the heterokaryons that recapitulated ontogeny, with early mesodermal factors being expressed before mature endothelial genes. A set of transcriptional factors not known to be involved in endothelial development was upregulated, one of which was POU class 3 homeobox 2 (Pou3f2). We confirmed its importance in differentiation to endothelial lineage via loss- and gain-of-function (LOF and GOF). Its role in vascular development was validated in zebrafish embryos using morpholino oligonucleotides. These studies provide a systematic and mechanistic approach for identifying key regulators in directed differentiation of pluripotent stem cells to somatic cell lineages.

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The following data sets were generated

Article and author information

Author details

  1. Wing Tak Wong

    Department of Cardiovascular Sciences, Houston Methodist Research Institute, Houston, United States
    Competing interests
    The authors declare that no competing interests exist.
  2. Gianfranco Matrone

    Department of Cardiovascular Sciences, Houston Methodist Research Institute, Houston, United States
    Competing interests
    The authors declare that no competing interests exist.
  3. XiaoYu Tian

    Department of Cardiovascular Sciences, Houston Methodist Research Institute, Houston, United States
    Competing interests
    The authors declare that no competing interests exist.
  4. Simion Alin Tomoiaga

    Department of Cardiovascular Sciences, Houston Methodist Research Institute, Houston, United States
    Competing interests
    The authors declare that no competing interests exist.
  5. Kin Fai Au

    Department of Internal Medicine, University of Iowa, Iowa City, United States
    Competing interests
    The authors declare that no competing interests exist.
  6. Shu Meng

    Department of Cardiovascular Sciences, Houston Methodist Research Institute, Houston, United States
    Competing interests
    The authors declare that no competing interests exist.
  7. Sayumi Yamazoe

    Department of Chemical and Systems Biology, Stanford University School of Medicine, Stanford, United States
    Competing interests
    The authors declare that no competing interests exist.
  8. Daniel Sieveking

    Department of Chemical and Systems Biology, Stanford University School of Medicine, Stanford, United States
    Competing interests
    The authors declare that no competing interests exist.
  9. Kaifu Chen

    Department of Cardiovascular Sciences, Houston Methodist Research Institute, Houston, United States
    Competing interests
    The authors declare that no competing interests exist.
  10. David M Burns

    Baxter Laboratory for Stem Cell Biology, Stanford University School of Medicine, Stanford, United States
    Competing interests
    The authors declare that no competing interests exist.
  11. James K Chen

    Department of Chemical and Systems Biology, Stanford University School of Medicine, Stanford, United States
    Competing interests
    The authors declare that no competing interests exist.
  12. Helen M Blau

    Baxter Laboratory for Stem Cell Biology, Stanford University School of Medicine, Stanford, United States
    Competing interests
    The authors declare that no competing interests exist.
  13. John P Cooke

    Department of Cardiovascular Sciences, Houston Methodist Research Institute, Houston, United States
    For correspondence
    jpcooke@houstonmethodist.org
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0003-0033-9138

Funding

National Institutes of Health

  • John P Cooke

American Heart Association

  • Wing Tak Wong

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

Ethics

Animal experimentation: Zebrafish are kept according to the laboratory protocols described in Zebrafish: A Practical Approach (Oxford University Press, 2002). These protocols comply with the Guide for the Care and Use of Laboratory Animals, the American Association for the Accreditation of Laboratory Animal Care (AAALAC) standards, and the regulations set forth in the Animals Welfare Act (P.L. 89-544, as amended by P.L. 91-579 and P.L . 94-279). Veterinary care is provided on a 24 hours basis, including weekends and holidays, by a staff of veterinarians with specialties in laboratory animal medicine and anesthesiology, and licensed animal health technicians. Training classes are offered. All veterinary care is provided by Houston Methodist Research Institute, which is fully accredited by AAALAC (ID A4555-01) and holds an approved NIH Assurance and USDA License (start date 03/08/2013). Support includes quarantine rooms, sterile operating rooms, post-surgical recovery rooms, radiology and diagnostic laboratory services. All surgery procedures were performed under anesthesia with Tricaine 0.02 mg/ml.

Reviewing Editor

  1. Gordana Vunjak-Novakovic, Columbia University, United States

Publication history

  1. Received: November 23, 2016
  2. Accepted: March 17, 2017
  3. Accepted Manuscript published: March 21, 2017 (version 1)
  4. Version of Record published: April 13, 2017 (version 2)
  5. Version of Record updated: April 24, 2017 (version 3)

Copyright

© 2017, Wong 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. Wing Tak Wong
  2. Gianfranco Matrone
  3. XiaoYu Tian
  4. Simion Alin Tomoiaga
  5. Kin Fai Au
  6. Shu Meng
  7. Sayumi Yamazoe
  8. Daniel Sieveking
  9. Kaifu Chen
  10. David M Burns
  11. James K Chen
  12. Helen M Blau
  13. John P Cooke
(2017)
Discovery of novel determinants of endothelial lineage using chimeric heterokaryons
eLife 6:e23588.
https://doi.org/10.7554/eLife.23588

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