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Patient-specific genomics and cross-species functional analysis implicate LRP2 in hypoplastic left heart syndrome

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Cite this article as: eLife 2020;9:e59554 doi: 10.7554/eLife.59554

Abstract

Congenital heart diseases (CHDs), including hypoplastic left heart syndrome (HLHS), are genetically complex and poorly understood. Here, a multi-disciplinary platform was established to functionally evaluate novel CHD gene candidates, based on whole genome and iPSC RNA sequencing of a HLHS family-trio. Filtering for rare variants and altered expression in proband iPSCs prioritized 10 candidates. siRNA/RNAi-mediated knockdown in generic human iPSC-derived cardiomyocytes (hiPSC-CM) and in developing Drosophila and zebrafish hearts revealed that LDL receptor-related protein LRP2 is required for cardiomyocyte proliferation and differentiation. Consistent with hypoplastic heart defects, compared to patents the proband's iPSC-CMs exhibited reduced proliferation. Interestingly, rare, predicted-damaging LRP2 variants were enriched in a HLHS cohort; however, understanding their contribution to HLHS requires further investigation. Collectively, we have established a multi-species high-throughput platform to rapidly evaluate candidate genes and their interactions during heart development, which are crucial first steps towards deciphering oligogenic underpinnings of CHDs, including maladaptive left hearts.

Article and author information

Author details

  1. Jeanne L Theis

    Cardiovascular Genetics Research Laboratory, Mayo Clinic, Rochester, United States
    Competing interests
    The authors declare that no competing interests exist.
  2. Georg Vogler

    Development, Aging and Regeneration, Sanford Burnham Prebys Medical Discovery Institute, La Jolla, United States
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-8303-3531
  3. Maria A Missinato

    Development, Aging and Regeneration, Sanford Burnham Prebys Medical Discovery Institute, La Jolla, United States
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-9055-758X
  4. Xing Li

    Division of Biomedical Statistics and Informatics, Mayo Clinic, Rochester, United States
    Competing interests
    The authors declare that no competing interests exist.
  5. Tanja Nielsen

    Development, Aging and Regeneration, Sanford Burnham Prebys Medical Discovery Institute, La Jolla, United States
    Competing interests
    The authors declare that no competing interests exist.
  6. Xin-Xin I Zeng

    Development, Aging and Regeneration, Sanford Burnham Prebys Medical Discovery Institute, La Jolla, United States
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-2707-7759
  7. Almudena Martinez-Fernandez

    Department of Cardiovascular Medicine, Mayo Clinic, Rochester, United States
    Competing interests
    The authors declare that no competing interests exist.
  8. Stanley M Walls

    Development, Aging and Regeneration, Sanford Burnham Prebys Medical Discovery Institute, La Jolla, United States
    Competing interests
    The authors declare that no competing interests exist.
  9. Anaïs Kervadec

    Development, Aging and Regeneration, Sanford Burnham Prebys Medical Discovery Institute, La Jolla, United States
    Competing interests
    The authors declare that no competing interests exist.
  10. James N Kezos

    Development, Aging and Regeneration, Sanford Burnham Prebys Medical Discovery Institute, La Jolla, United States
    Competing interests
    The authors declare that no competing interests exist.
  11. Katja Birker

    Development, Aging and Regeneration, Sanford Burnham Prebys Medical Discovery Institute, La Jolla, United States
    Competing interests
    The authors declare that no competing interests exist.
  12. Jared M Evans

    Division of Biomedical Statistics and Informatics, Mayo Clinic, Rochester, United States
    Competing interests
    The authors declare that no competing interests exist.
  13. Megan M O'Byrne

    Division of Biomedical Statistics and Informatics, Mayo Clinic, Rochester, United States
    Competing interests
    The authors declare that no competing interests exist.
  14. Zachary C Fogarty

    Division of Biomedical Statistics and Informatics, Mayo Clinic, Rochester, United States
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-5588-3216
  15. André Terzic

    Department of Cardiovascular Medicine, Mayo Clinic, Rochester, United States
    Competing interests
    The authors declare that no competing interests exist.
  16. Paul Grossfeld

    Rady's Hospital, University of California San Diego, San Diego, United States
    Competing interests
    The authors declare that no competing interests exist.
  17. Karen Ocorr

    Development, Aging and Regeneration, Sanford Burnham Prebys Medical Discovery Institute, La Jolla, United States
    Competing interests
    The authors declare that no competing interests exist.
  18. Timothy J Nelson

    Department of Internal Medicine, Mayo Clinic, Rochester, United States
    For correspondence
    nelson.timothy@mayo.edu
    Competing interests
    The authors declare that no competing interests exist.
  19. Timothy M Olson

    Department of Cardiovascular Medicine, Mayo Clinic, Rochester, United States
    For correspondence
    olson.timothy@mayo.edu
    Competing interests
    The authors declare that no competing interests exist.
  20. Alexandre R Colas

    Development, Aging and Regeneration, Sanford Burnham Prebys Medical Discovery Institute, La Jolla, United States
    Competing interests
    The authors declare that no competing interests exist.
  21. Rolf Bodmer

    Development, Aging, and Regeneration Program, Sanford Burnham Prebys Medical Discovery Institute, La Jolla, United States
    For correspondence
    rolf@sbpdiscovery.org
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-9087-1210

Funding

Todd and Karen Wanek Family Program for Hypoplastic Left Heart Syndrome, Mayo Clinic Foundation, Rochester, MN (SAN-233970)

  • Alexandre R Colas
  • Rolf Bodmer

National Institutes of Health (HL054732)

  • Rolf Bodmer

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

Ethics

Animal experimentation: SBP has retained the services of a veterinarian who is a diplomat of the American College of Laboratory Animal Medicine. Close contact with Animal Facility personnel is maintained through telephone calls and on-campus visits once a week. This person is a member of the Institute's Animal Care and Use Committee (IACUC)and attends monthly meetings. This study was performed in strict accordance with the recommendations in the Guide for the Care and Use of Laboratory Animals of the National Institutes of Health. All of the animals were handled according to the Institute's Animal Care and Use Program, which is accredited by AAALAC International, and a Multiple Project Assurance A3053-1 is on file in the OLAW, DHHS. The protocol was approved by SBP IACUC (Permit Number: 19-087). Animals are euthanized after filming hearts by an overdose of anesthetic (3-aminobenzoic acid ethyl ester (MS-222) at 250-300 mg/L.

Human subjects: Written informed consent was obtained for the index family and an HLHS cohort, under a research protocol approved by the Mayo Clinic Institutional Review Board (11-000114 "Genetic Investigations in Hypoplastic Left Heart Syndrome").

Reviewing Editor

  1. Antje Ebert

Publication history

  1. Received: June 1, 2020
  2. Accepted: September 29, 2020
  3. Accepted Manuscript published: October 2, 2020 (version 1)
  4. Version of Record published: October 22, 2020 (version 2)
  5. Version of Record updated: November 23, 2020 (version 3)

Copyright

© 2020, Theis 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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