A1 Refereed original research article in a scientific journal
The ERBB2 c.1795C>T, p.Arg599Cys variant is associated with left ventricular outflow tract obstruction defects in humans
Authors: Ampuja, Minna; Ericsson, Sabina; Paatero, Ilkka; Chowdhury, Iftekhar; Villman, Jenna; Broberg, Martin; Ramste, Amanda; Balboa, Diego; Ojala, Tiina; Chong, Jessica X.; Bamshad, Michael J.; Priest, James R.; Varjosalo, Markku; Kivelä, Riikka; Helle, Emmi
Publisher: ELSEVIER
Publishing place: AMSTERDAM
Publication year: 2025
Journal: HGG advances
Journal name in source: HUMAN GENETICS AND GENOMICS ADVANCES
Journal acronym: HUM GENET GENOM ADV
Article number: 100446
Volume: 6
Issue: 3
Number of pages: 15
eISSN: 2666-2477
DOI: https://doi.org/10.1016/j.xhgg.2025.100446
Web address : https://doi.org/10.1016/j.xhgg.2025.100446
Self-archived copy’s web address: https://research.utu.fi/converis/portal/detail/Publication/492315921
Non-syndromic congenital heart defects (CHDs) are occasionally familial and left ventricular outflow tract obstruction (LVOTO) defects are among the subtypes with the highest hereditability. The aim of this study was to evaluate the pathogenicity of a heterozygous ERBB2 variant c.1795C>T, p.Arg599Cys identified in three families with LVOTO defects. Variant detection was done with exome sequencing. Western blotting, digital PCR, mass spectrometry (MS), MS microscopy, and flow cytometry were used to study the function of the ERBB2 variant c.1795C>T. Cardiac structure and function were studied in zebrafish embryos expressing human ERBB2 wild type or c.1795C>T. Proband-derived human induced pluripotent stem cell cardiomyocytes (hiPS-CMs) and endothelial cells (hiPS-ECs) were used for transcriptomic analyses. While phosphorylation of the ERBB2 p.Arg599Cys receptor was not altered, the variant affected dramatically the binding partners of the protein, indicating mislocalization of the mutant ERBB2 from plasma membrane to endoplasmic reticulum. Expression of human ERBB2 p.Arg599Cys in zebrafish embryos resulted in cardiomyocyte hypertrophy, increased cardiac wall thickness, and impaired fractional shortening. Transcriptomic analyses of hiPS-ECs and hiPS-CMs from an individual with the c.1795C>T variant showed aberrant expression of genes related to cardiovascular system development and abnormal response to oxidative stress in both cell types. In conclusion, the heterozygous variant ERBB2 c.1795C>T, p.Arg599Cys leads to abnormal cellular localization of the ERBB2 receptor and induces structural changes and dysfunction in the zebrafish embryo heart. This evidence expands previous findings from animal studies to humans and suggests variants in ERBB2 may be associated with CHD.
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Funding information in the publication:
This study has been funded by the Academy of Finland (297245, 331405), the Finnish Medical Foundation, Foundation for Pediatric Research (Helsinki, Finland), The Finnish Cultural Foundation (Helsinki, Finland), Finnish Foundation for Cardiovascular Research (Helsinki, Finland), Sigrid Juselius Foundation, University of Helsinki, Helsinki University Hospital, Orion Research Foundation, Päivikki and Sakari Sohlberg Foundation, and Aarne Koskelo Foundation. Exome sequencing and data analysis were provided by the University of Washington Center for Rare Disease Research (UW-CRDR) with support from NHGRI grants U01 HG011744, UM1 HG006493, and U24 HG011746. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health. We thank Ilse Paetau and Katja Salo for their help in laboratory experiments and administrative support. We thank Kari Alitalo, Tanja Laakkonen, and Seppo Kaijalainen for help with the ERBB2 c.1795C>T plasmid. We thank Professor Anu Suomalainen-Wartiovaara, Professor Timo Otonkoski, and Docent Ras Trokovic for the iPS cells. We thank Michael Jeltsch for providing VEGF for the experiments. We thank Zebrafish Core and Cell Imaging Core (both Turku Bioscience Center, University of Turku, and Åbo Akademi University, supported by Biocenter Finland) for instrumentation and services. scRNA-seq service was performed at FIMM Single-Cell Analytics unit, RNA-seq at Biomedicum Functional Genomics Unit and imaging at the Biomedicum Imaging Unit, all supported by HiLIFE and Biocenter Finland at the University of Helsinki. We acknowledge the HiLife Flow Cytometry Unit, University of Helsinki and Biomedicum Virus Core Unit (BVC), University of Helsinki.
