Missense and truncated variants in ERF in individuals with a Noonan-like phenotype without craniosynostosis

Abstract ETS2 repressor factor (ERF) is a member of the ETS family of transcriptional repressors downstream of ERK. Although germline truncated variants in ERF have been identified in individuals with Noonan-like syndrome with or without craniosynostosis, the clinical spectrum of ERF variant-positiv...

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Main Authors: Yusuke Goto, Tetsuya Niihori, Seiji Mizuno, Nobuhiko Okamoto, Tsutomu Ogata, Kenji Kurosawa, Hirofumi Ohashi, Yoichi Matsubara, Taiki Abe, Atsuo Kikuchi, Yoko Aoki
Format: Article
Language:English
Published: Nature Portfolio 2025-04-01
Series:Scientific Reports
Online Access:https://doi.org/10.1038/s41598-025-89719-1
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author Yusuke Goto
Tetsuya Niihori
Seiji Mizuno
Nobuhiko Okamoto
Tsutomu Ogata
Kenji Kurosawa
Hirofumi Ohashi
Yoichi Matsubara
Taiki Abe
Atsuo Kikuchi
Yoko Aoki
author_facet Yusuke Goto
Tetsuya Niihori
Seiji Mizuno
Nobuhiko Okamoto
Tsutomu Ogata
Kenji Kurosawa
Hirofumi Ohashi
Yoichi Matsubara
Taiki Abe
Atsuo Kikuchi
Yoko Aoki
author_sort Yusuke Goto
collection DOAJ
description Abstract ETS2 repressor factor (ERF) is a member of the ETS family of transcriptional repressors downstream of ERK. Although germline truncated variants in ERF have been identified in individuals with Noonan-like syndrome with or without craniosynostosis, the clinical spectrum of ERF variant-positive individuals and the functional characterization of ERF variants are currently not fully understood. In this study, we identified one missense variant (p.G53R) and two truncating variants in ERF using whole exome sequencing (WES) in three individuals and one truncating variant using Sanger sequencing in one of 81 individuals with suspected Noonan syndrome without any pathogenic variants by targeted analysis in the previous study. Four Individuals with pathogenic ERF variants were diagnosed with Noonan-like syndrome, where craniosynostosis was not evident. Our investigation revealed that wild-type ERF undergoes nuclear-cytoplasmic shift, whereas truncated mutant ERF are predominantly localized in the nucleus. Moreover, R183* and G299Rfs variants lost their ability to repress the proliferation of osteoblast-like cells (MC3T3-E1). A luciferase assay examining the transcriptional activity of RUNX2 binding motifs indicated that the truncated variants were defective in their suppressive function. Further experimentation demonstrated that MC3T3-E1 cells expressing the p.G53R and three truncating variants induced ossification compared to the wild-type. These results suggest that loss-of-function mutations in ERF, which result in reduced ossification suppressor activity in MC3T3-E1 cells, can lead to craniofacial abnormalities in individuals with Noonan syndrome-like symptoms.
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spelling doaj-art-41d4015cab9d4eb0bd6e343daa6633b32025-08-20T02:10:50ZengNature PortfolioScientific Reports2045-23222025-04-0115111510.1038/s41598-025-89719-1Missense and truncated variants in ERF in individuals with a Noonan-like phenotype without craniosynostosisYusuke Goto0Tetsuya Niihori1Seiji Mizuno2Nobuhiko Okamoto3Tsutomu Ogata4Kenji Kurosawa5Hirofumi Ohashi6Yoichi Matsubara7Taiki Abe8Atsuo Kikuchi9Yoko Aoki10Department of Medical Genetics, Tohoku University School of MedicineDepartment of Medical Genetics, Tohoku University School of MedicineDepartment of Pediatrics, Central Hospital, Aichi Developmental Disability CenterDepartment of Medical Genetics, Research Institute, Osaka Women’s and Children’s HospitalDepartment of Pediatrics, Hamamatsu Medical CenterDivision of Medical Genetics, Kanagawa Children’s Medical CenterDivision of Medical Genetics, Saitama Children’s Medical CenterNational Center for Child Health and DevelopmentDepartment of Medical Genetics, Tohoku University School of MedicineDepartment of Pediatrics, Tohoku University School of MedicineDepartment of Medical Genetics, Tohoku University School of MedicineAbstract ETS2 repressor factor (ERF) is a member of the ETS family of transcriptional repressors downstream of ERK. Although germline truncated variants in ERF have been identified in individuals with Noonan-like syndrome with or without craniosynostosis, the clinical spectrum of ERF variant-positive individuals and the functional characterization of ERF variants are currently not fully understood. In this study, we identified one missense variant (p.G53R) and two truncating variants in ERF using whole exome sequencing (WES) in three individuals and one truncating variant using Sanger sequencing in one of 81 individuals with suspected Noonan syndrome without any pathogenic variants by targeted analysis in the previous study. Four Individuals with pathogenic ERF variants were diagnosed with Noonan-like syndrome, where craniosynostosis was not evident. Our investigation revealed that wild-type ERF undergoes nuclear-cytoplasmic shift, whereas truncated mutant ERF are predominantly localized in the nucleus. Moreover, R183* and G299Rfs variants lost their ability to repress the proliferation of osteoblast-like cells (MC3T3-E1). A luciferase assay examining the transcriptional activity of RUNX2 binding motifs indicated that the truncated variants were defective in their suppressive function. Further experimentation demonstrated that MC3T3-E1 cells expressing the p.G53R and three truncating variants induced ossification compared to the wild-type. These results suggest that loss-of-function mutations in ERF, which result in reduced ossification suppressor activity in MC3T3-E1 cells, can lead to craniofacial abnormalities in individuals with Noonan syndrome-like symptoms.https://doi.org/10.1038/s41598-025-89719-1
spellingShingle Yusuke Goto
Tetsuya Niihori
Seiji Mizuno
Nobuhiko Okamoto
Tsutomu Ogata
Kenji Kurosawa
Hirofumi Ohashi
Yoichi Matsubara
Taiki Abe
Atsuo Kikuchi
Yoko Aoki
Missense and truncated variants in ERF in individuals with a Noonan-like phenotype without craniosynostosis
Scientific Reports
title Missense and truncated variants in ERF in individuals with a Noonan-like phenotype without craniosynostosis
title_full Missense and truncated variants in ERF in individuals with a Noonan-like phenotype without craniosynostosis
title_fullStr Missense and truncated variants in ERF in individuals with a Noonan-like phenotype without craniosynostosis
title_full_unstemmed Missense and truncated variants in ERF in individuals with a Noonan-like phenotype without craniosynostosis
title_short Missense and truncated variants in ERF in individuals with a Noonan-like phenotype without craniosynostosis
title_sort missense and truncated variants in erf in individuals with a noonan like phenotype without craniosynostosis
url https://doi.org/10.1038/s41598-025-89719-1
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