EAP1 regulation of GnRH promoter activity is important for human pubertal timing

Alessandra Mancini, Sasha R. Howard, Claudia P. Cabrera, Michael R. Barnes, Alessia David, Karoliina Wehkalampi, Sabine Heger, Alejandro Lomniczi, Leonardo Guasti, Sergio Ojeda, Leo Dunkel

    Research output: Contribution to journalArticle

    2 Citations (Scopus)

    Abstract

    The initiation of puberty is orchestrated by an augmentation of gonadotropin-releasing hormone (GnRH) secretion from a few thousand hypothalamic neurons. Recent findings have indicated that the neuroendocrine control of puberty may be regulated by a hierarchically organized network of transcriptional factors acting upstream of GnRH. These include enhanced at puberty 1 (EAP1), which contributes to the initiation of female puberty through transactivation of the GnRH promoter. However, no EAP1 mutations have been found in humans with disorders of pubertal timing. We performed whole-exome sequencing in 67 probands and 93 relatives from a large cohort of familial self-limited delayed puberty (DP). Variants were analyzed for rare, potentially pathogenic variants enriched in case versus controls and relevant to the biological control of puberty. We identified one in-frame deletion (Ala221del) and one rare missense variant (Asn770His) in EAP1 in two unrelated families; these variants were highly conserved and potentially pathogenic. Expression studies revealed Eap1 mRNA abundance in peri-pubertal mouse hypothalamus. EAP1 binding to the GnRH1 promoter increased in monkey hypothalamus at the onset of puberty as determined by chromatin immunoprecipitation. Using a luciferase reporter assay, EAP1 mutants showed a reduced ability to trans-activate the GnRH promoter compared to wild-type EAP1, due to reduced protein levels caused by the Ala221del mutation and subcellular mislocation caused by the Asn770His mutation, as revealed by western blot and immunofluorescence, respectively. In conclusion, we have identified the first EAP1 mutations leading to reduced GnRH transcriptional activity resulting in a phenotype of self-limited DP.

    Original languageEnglish (US)
    Pages (from-to)1357-1368
    Number of pages12
    JournalHuman molecular genetics
    Volume28
    Issue number8
    DOIs
    StatePublished - Apr 15 2019

    Fingerprint

    Puberty
    Gonadotropin-Releasing Hormone
    Delayed Puberty
    Mutation
    Hypothalamus
    Exome
    Gene Regulatory Networks
    Chromatin Immunoprecipitation
    Luciferases
    Transcriptional Activation
    Haplorhini
    Fluorescent Antibody Technique
    Western Blotting
    Phenotype
    Neurons

    ASJC Scopus subject areas

    • Molecular Biology
    • Genetics
    • Genetics(clinical)

    Cite this

    Mancini, A., Howard, S. R., Cabrera, C. P., Barnes, M. R., David, A., Wehkalampi, K., ... Dunkel, L. (2019). EAP1 regulation of GnRH promoter activity is important for human pubertal timing. Human molecular genetics, 28(8), 1357-1368. https://doi.org/10.1093/hmg/ddy451

    EAP1 regulation of GnRH promoter activity is important for human pubertal timing. / Mancini, Alessandra; Howard, Sasha R.; Cabrera, Claudia P.; Barnes, Michael R.; David, Alessia; Wehkalampi, Karoliina; Heger, Sabine; Lomniczi, Alejandro; Guasti, Leonardo; Ojeda, Sergio; Dunkel, Leo.

    In: Human molecular genetics, Vol. 28, No. 8, 15.04.2019, p. 1357-1368.

    Research output: Contribution to journalArticle

    Mancini, A, Howard, SR, Cabrera, CP, Barnes, MR, David, A, Wehkalampi, K, Heger, S, Lomniczi, A, Guasti, L, Ojeda, S & Dunkel, L 2019, 'EAP1 regulation of GnRH promoter activity is important for human pubertal timing', Human molecular genetics, vol. 28, no. 8, pp. 1357-1368. https://doi.org/10.1093/hmg/ddy451
    Mancini A, Howard SR, Cabrera CP, Barnes MR, David A, Wehkalampi K et al. EAP1 regulation of GnRH promoter activity is important for human pubertal timing. Human molecular genetics. 2019 Apr 15;28(8):1357-1368. https://doi.org/10.1093/hmg/ddy451
    Mancini, Alessandra ; Howard, Sasha R. ; Cabrera, Claudia P. ; Barnes, Michael R. ; David, Alessia ; Wehkalampi, Karoliina ; Heger, Sabine ; Lomniczi, Alejandro ; Guasti, Leonardo ; Ojeda, Sergio ; Dunkel, Leo. / EAP1 regulation of GnRH promoter activity is important for human pubertal timing. In: Human molecular genetics. 2019 ; Vol. 28, No. 8. pp. 1357-1368.
    @article{94e5e862843d4a3aa682758f40240565,
    title = "EAP1 regulation of GnRH promoter activity is important for human pubertal timing",
    abstract = "The initiation of puberty is orchestrated by an augmentation of gonadotropin-releasing hormone (GnRH) secretion from a few thousand hypothalamic neurons. Recent findings have indicated that the neuroendocrine control of puberty may be regulated by a hierarchically organized network of transcriptional factors acting upstream of GnRH. These include enhanced at puberty 1 (EAP1), which contributes to the initiation of female puberty through transactivation of the GnRH promoter. However, no EAP1 mutations have been found in humans with disorders of pubertal timing. We performed whole-exome sequencing in 67 probands and 93 relatives from a large cohort of familial self-limited delayed puberty (DP). Variants were analyzed for rare, potentially pathogenic variants enriched in case versus controls and relevant to the biological control of puberty. We identified one in-frame deletion (Ala221del) and one rare missense variant (Asn770His) in EAP1 in two unrelated families; these variants were highly conserved and potentially pathogenic. Expression studies revealed Eap1 mRNA abundance in peri-pubertal mouse hypothalamus. EAP1 binding to the GnRH1 promoter increased in monkey hypothalamus at the onset of puberty as determined by chromatin immunoprecipitation. Using a luciferase reporter assay, EAP1 mutants showed a reduced ability to trans-activate the GnRH promoter compared to wild-type EAP1, due to reduced protein levels caused by the Ala221del mutation and subcellular mislocation caused by the Asn770His mutation, as revealed by western blot and immunofluorescence, respectively. In conclusion, we have identified the first EAP1 mutations leading to reduced GnRH transcriptional activity resulting in a phenotype of self-limited DP.",
    author = "Alessandra Mancini and Howard, {Sasha R.} and Cabrera, {Claudia P.} and Barnes, {Michael R.} and Alessia David and Karoliina Wehkalampi and Sabine Heger and Alejandro Lomniczi and Leonardo Guasti and Sergio Ojeda and Leo Dunkel",
    year = "2019",
    month = "4",
    day = "15",
    doi = "10.1093/hmg/ddy451",
    language = "English (US)",
    volume = "28",
    pages = "1357--1368",
    journal = "Human Molecular Genetics",
    issn = "0964-6906",
    publisher = "Oxford University Press",
    number = "8",

    }

    TY - JOUR

    T1 - EAP1 regulation of GnRH promoter activity is important for human pubertal timing

    AU - Mancini, Alessandra

    AU - Howard, Sasha R.

    AU - Cabrera, Claudia P.

    AU - Barnes, Michael R.

    AU - David, Alessia

    AU - Wehkalampi, Karoliina

    AU - Heger, Sabine

    AU - Lomniczi, Alejandro

    AU - Guasti, Leonardo

    AU - Ojeda, Sergio

    AU - Dunkel, Leo

    PY - 2019/4/15

    Y1 - 2019/4/15

    N2 - The initiation of puberty is orchestrated by an augmentation of gonadotropin-releasing hormone (GnRH) secretion from a few thousand hypothalamic neurons. Recent findings have indicated that the neuroendocrine control of puberty may be regulated by a hierarchically organized network of transcriptional factors acting upstream of GnRH. These include enhanced at puberty 1 (EAP1), which contributes to the initiation of female puberty through transactivation of the GnRH promoter. However, no EAP1 mutations have been found in humans with disorders of pubertal timing. We performed whole-exome sequencing in 67 probands and 93 relatives from a large cohort of familial self-limited delayed puberty (DP). Variants were analyzed for rare, potentially pathogenic variants enriched in case versus controls and relevant to the biological control of puberty. We identified one in-frame deletion (Ala221del) and one rare missense variant (Asn770His) in EAP1 in two unrelated families; these variants were highly conserved and potentially pathogenic. Expression studies revealed Eap1 mRNA abundance in peri-pubertal mouse hypothalamus. EAP1 binding to the GnRH1 promoter increased in monkey hypothalamus at the onset of puberty as determined by chromatin immunoprecipitation. Using a luciferase reporter assay, EAP1 mutants showed a reduced ability to trans-activate the GnRH promoter compared to wild-type EAP1, due to reduced protein levels caused by the Ala221del mutation and subcellular mislocation caused by the Asn770His mutation, as revealed by western blot and immunofluorescence, respectively. In conclusion, we have identified the first EAP1 mutations leading to reduced GnRH transcriptional activity resulting in a phenotype of self-limited DP.

    AB - The initiation of puberty is orchestrated by an augmentation of gonadotropin-releasing hormone (GnRH) secretion from a few thousand hypothalamic neurons. Recent findings have indicated that the neuroendocrine control of puberty may be regulated by a hierarchically organized network of transcriptional factors acting upstream of GnRH. These include enhanced at puberty 1 (EAP1), which contributes to the initiation of female puberty through transactivation of the GnRH promoter. However, no EAP1 mutations have been found in humans with disorders of pubertal timing. We performed whole-exome sequencing in 67 probands and 93 relatives from a large cohort of familial self-limited delayed puberty (DP). Variants were analyzed for rare, potentially pathogenic variants enriched in case versus controls and relevant to the biological control of puberty. We identified one in-frame deletion (Ala221del) and one rare missense variant (Asn770His) in EAP1 in two unrelated families; these variants were highly conserved and potentially pathogenic. Expression studies revealed Eap1 mRNA abundance in peri-pubertal mouse hypothalamus. EAP1 binding to the GnRH1 promoter increased in monkey hypothalamus at the onset of puberty as determined by chromatin immunoprecipitation. Using a luciferase reporter assay, EAP1 mutants showed a reduced ability to trans-activate the GnRH promoter compared to wild-type EAP1, due to reduced protein levels caused by the Ala221del mutation and subcellular mislocation caused by the Asn770His mutation, as revealed by western blot and immunofluorescence, respectively. In conclusion, we have identified the first EAP1 mutations leading to reduced GnRH transcriptional activity resulting in a phenotype of self-limited DP.

    UR - http://www.scopus.com/inward/record.url?scp=85064527607&partnerID=8YFLogxK

    UR - http://www.scopus.com/inward/citedby.url?scp=85064527607&partnerID=8YFLogxK

    U2 - 10.1093/hmg/ddy451

    DO - 10.1093/hmg/ddy451

    M3 - Article

    VL - 28

    SP - 1357

    EP - 1368

    JO - Human Molecular Genetics

    JF - Human Molecular Genetics

    SN - 0964-6906

    IS - 8

    ER -