Abstract
Heterozygous deleterious null alleles and specific missense variants in the DNA-binding domain of the ETS2 repressor factor (ERF) cause craniosynostosis, while the recurrent p.(Tyr89Cys) missense variant is associated with Chitayat syndrome. Exome and whole transcriptome sequencing revealed the ERF de novo in-frame indel c.911_913del selectively removing the serine of the FSF motif, which interacts with the extracellular signal-regulated kinases (ERKs), in a 10-year-old girl with microcephaly, multiple congenital joint dislocations, generalized joint hypermobility, and Pierre-Robin sequence. Three additional cases with developmental delay variably associated with microcephaly, Pierre-Robin sequence and minor skeletal anomalies were detected carrying heterozygous de novo non-truncating alleles (two with c.911_913del and one with the missense c.907 T > A change) in the same FSF motif. Protein affinity maps, co-immunoprecipitation experiments and subcellular distribution showed that both the variants impair the interaction between ERF and activated ERK1/2 and increase ERF nuclear localization, affecting ERF repressor activity that may lead to developmental defects. Our work expands the phenotypic spectrum of ERF-related disorders to a pleiotropic condition with microcephaly, developmental delay and skeletal anomalies, that we termed MIDES syndrome, and adds to the understanding of the relevance of the ERF-ERK interaction in human development and disease.
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Fig. 1: Clinical and molecular data of Individual 1.
Fig. 2: ERF-Phe303Ile and ERF-Ser304del variants alter the ability of ERF to interact with the ERK1/2 kinases.
Fig. 3: ERF-Phe303Ile and ERF-Ser304del variants strongly decrease interaction with activated ERK1/2 and increase nuclear accumulation of ERF.
Data availability
The authors declare that all data supporting the findings of this study are available within the article and its supplementary material files, or from the corresponding author upon reasonable request.
Code availability
The underlying code for this study may be made available to qualified researchers on reasonable request from the corresponding author.
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Acknowledgements
MC is member of the ReCONNET ERN (European Reference Network for rare and complex musculoskeletal diseases) and ERN SKIN (European Reference Network for rare and complex skin diseases).
Funding
This work was supported by the Ricerca Corrente 2022–2024 to MC, Ricerca Finalizzata (project code: RF_2021_12373524), Italian Ministry of Health to MC and DC; European Regional Development Fund of the European Union and Greek national funds through the National Recovery and Resilience Program (project code: TAEDR-0535850) to GM; Fondazione Telethon Core Grant, Armenise-Harvard Foundation Career Development Award, European Research Council (grant agreement 759154, CellKarma) and Piano Operativo Salute Traiettoria 3, “Genomed,” Italian Ministry of Health to DC. The funder played no role in study design, data collection, analysis and interpretation of data, or the writing of this manuscript.
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Authors and Affiliations
Division of Medical Genetics, Fondazione IRCCS-Casa Sollievo della Sofferenza, Viale Cappuccini snc, 71013, San Giovanni Rotondo, Italy
Lucia Micale, Carmela Fusco, Riccardo Pracella, Grazia Nardella, Maria Pia Leone & Marco Castori
IMBB, FORTH, 71003, Heraklion, Crete, Greece
Aikaterini Vourlia, Lydia Xenou & George Mavrothalassitis
Medical School, University of Crete, 71003, Heraklion, Crete, Greece
Aikaterini Vourlia, Dimitrios-Christoforos Karagiannis & George Mavrothalassitis
Armenise/Harvard Laboratory of Integrative Genomics, Telethon Institute of Genetics and Medicine (TIGEM), Via Campi Flegrei 34, 80078, Pozzuoli, Italy
Lorenzo Vaccaro, Antonio Gramazio & Davide Cacchiarelli
Department of Translational Medicine, University of Naples “Federico II”, Naples, Italy
Lorenzo Vaccaro, Antonio Gramazio & Davide Cacchiarelli
Rare Diseases and Medical Genetics, IRCCS Ospedale Pediatrico Bambino Gesù, Rome, Italy
Maria Lisa Dentici
Translational Cytogenetics, IRCCS Ospedale Pediatrico Bambino Gesù, Rome, Italy
Chiara Aiello & Antonio Novelli
Department of Genome Sciences, University of Washington School of Medicine, Seattle, WA, USA
Yang Sui & Evan E. Eichler
Howard Hughes Medical Institute, University of Washington, Seattle, WA, USA
Evan E. Eichler
Genomics and Experimental Medicine Program, Scuola Superiore Meridionale, Naples, Italy
Davide Cacchiarelli
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Lucia Micale
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Contributions
MC, GM, and LM contributed to the study conception and design. Material preparation, data collection and interpretation were performed by MC, DC, GM, and LM. Funtional analysis were performed by CF, DCK, LM, GN, AV, and LX. ES data were provided by CA, EEE, MLD, MPL, LM, AN, and YS. RNA sequencing was carried out by AG, LV, and DC. In silico analysis was performed by RP. The first draft of the manuscript was written by MC, LM, and GM. All authors commented on previous versions of the manuscript. All authors read and approved the final manuscript.
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Correspondence to Lucia Micale or George Mavrothalassitis.
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All authors declare that there is no conflict of interest concerning this work. DC is founder, shareholder, and consultant of NEGEDIA S.r.l.
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This study is in accordance with the 1984 Helsinki declaration and subsequent revisions, and received IRB approval at Fondazione IRCCS-Casa Sollievo della Sofferenza (approval no. 2023/45/CE). All patients provided written informed consent for participation in this study and publication of molecular and clinical data.
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Micale, L., Vourlia, A., Fusco, C. et al. Heterozygous variants disrupting the interaction of ERF with activated ERK1/2 cause microcephaly, developmental delay, and skeletal anomalies. Eur J Hum Genet (2024). https://doi.org/10.1038/s41431-024-01721-9
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Received:18 June 2024
Revised:20 August 2024
Accepted:17 October 2024
Published:12 December 2024
DOI:https://doi.org/10.1038/s41431-024-01721-9
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