Estrogens enhance myoblast differentiation in facioscapulohumeral muscular dystrophy by antagonizing DUX4 activity

E. Teveroni, M. Pellegrino, S. Sacconi, P. Calandra, I. Cascino, S. Farioli-Vecchioli, A. Puma, M. Garibaldi, R. Morosetti, G. Tasca, E. Ricci, C.P. Trevisan, G. Galluzzi, A. Pontecorvi, M. Crescenzi, G. Deidda, F. Moretti

Research output: Contribution to journalArticle

13 Citations (Scopus)

Abstract

Facioscapulohumeral muscular dystrophy (FSHD) is an autosomal dominant neuromuscular disorder that is characterized by extreme variability in symptoms, with females being less severely affected than males and presenting a higher proportion of asymptomatic carriers. The sex-related factors involved in the disease are not known. Here, we have utilized myoblasts isolated from FSHD patients (FSHD myoblasts) to investigate the effect of estrogens on muscle properties. Our results demonstrated that estrogens counteract the differentiation impairment of FSHD myoblasts without affecting cell proliferation or survival. Estrogen effects are mediated by estrogen receptor ? (ER?), which reduces chromatin occupancy and transcriptional activity of double homeobox 4 (DUX4), a protein whose aberrant expression has been implicated in FSHD pathogenesis. During myoblast differentiation, we observed that the levels and activity of DUX4 increased progressively and were associated with its enhanced recruitment in the nucleus. ER? interfered with this recruitment by relocalizing DUX4 in the cytoplasm. This work identifies estrogens as a potential disease modifier that underlie sex-related differences in FSHD by protecting against myoblast differentiation impairments in this disease.
Original languageEnglish
Pages (from-to)1531-1545
Number of pages15
JournalJournal of Clinical Investigation
Volume127
Issue number4
DOIs
Publication statusPublished - 2017

Fingerprint

Facioscapulohumeral Muscular Dystrophy
Homeobox Genes
Myoblasts
Estrogens
Sex Factors
Sex Characteristics
Estrogen Receptors
Chromatin
Cell Survival
Cytoplasm
Cell Proliferation
Muscles

Keywords

  • double homeobox 4 protein
  • estrogen
  • estrogen receptor beta
  • homeodomain protein
  • unclassified drug
  • DUX4 protein, human
  • ESR2 protein, human
  • estradiol
  • Article
  • case report
  • cell differentiation
  • cell nucleus
  • cell proliferation
  • cell survival
  • cellular distribution
  • chromatin
  • controlled study
  • cytoplasm
  • facioscapulohumeral muscular dystrophy
  • female
  • human
  • human cell
  • male
  • myoblast
  • priority journal
  • protein expression
  • cell culture
  • gene expression
  • metabolism
  • pathology
  • physiology
  • protein transport
  • transcription initiation
  • Cell Differentiation
  • Cells, Cultured
  • Estradiol
  • Estrogen Receptor beta
  • Estrogens
  • Gene Expression
  • Homeodomain Proteins
  • Humans
  • Muscular Dystrophy, Facioscapulohumeral
  • Myoblasts
  • Protein Transport
  • Transcriptional Activation

Cite this

Teveroni, E., Pellegrino, M., Sacconi, S., Calandra, P., Cascino, I., Farioli-Vecchioli, S., ... Moretti, F. (2017). Estrogens enhance myoblast differentiation in facioscapulohumeral muscular dystrophy by antagonizing DUX4 activity. Journal of Clinical Investigation, 127(4), 1531-1545. https://doi.org/10.1172/JCI89401

Estrogens enhance myoblast differentiation in facioscapulohumeral muscular dystrophy by antagonizing DUX4 activity. / Teveroni, E.; Pellegrino, M.; Sacconi, S.; Calandra, P.; Cascino, I.; Farioli-Vecchioli, S.; Puma, A.; Garibaldi, M.; Morosetti, R.; Tasca, G.; Ricci, E.; Trevisan, C.P.; Galluzzi, G.; Pontecorvi, A.; Crescenzi, M.; Deidda, G.; Moretti, F.

In: Journal of Clinical Investigation, Vol. 127, No. 4, 2017, p. 1531-1545.

Research output: Contribution to journalArticle

Teveroni, E, Pellegrino, M, Sacconi, S, Calandra, P, Cascino, I, Farioli-Vecchioli, S, Puma, A, Garibaldi, M, Morosetti, R, Tasca, G, Ricci, E, Trevisan, CP, Galluzzi, G, Pontecorvi, A, Crescenzi, M, Deidda, G & Moretti, F 2017, 'Estrogens enhance myoblast differentiation in facioscapulohumeral muscular dystrophy by antagonizing DUX4 activity', Journal of Clinical Investigation, vol. 127, no. 4, pp. 1531-1545. https://doi.org/10.1172/JCI89401
Teveroni, E. ; Pellegrino, M. ; Sacconi, S. ; Calandra, P. ; Cascino, I. ; Farioli-Vecchioli, S. ; Puma, A. ; Garibaldi, M. ; Morosetti, R. ; Tasca, G. ; Ricci, E. ; Trevisan, C.P. ; Galluzzi, G. ; Pontecorvi, A. ; Crescenzi, M. ; Deidda, G. ; Moretti, F. / Estrogens enhance myoblast differentiation in facioscapulohumeral muscular dystrophy by antagonizing DUX4 activity. In: Journal of Clinical Investigation. 2017 ; Vol. 127, No. 4. pp. 1531-1545.
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title = "Estrogens enhance myoblast differentiation in facioscapulohumeral muscular dystrophy by antagonizing DUX4 activity",
abstract = "Facioscapulohumeral muscular dystrophy (FSHD) is an autosomal dominant neuromuscular disorder that is characterized by extreme variability in symptoms, with females being less severely affected than males and presenting a higher proportion of asymptomatic carriers. The sex-related factors involved in the disease are not known. Here, we have utilized myoblasts isolated from FSHD patients (FSHD myoblasts) to investigate the effect of estrogens on muscle properties. Our results demonstrated that estrogens counteract the differentiation impairment of FSHD myoblasts without affecting cell proliferation or survival. Estrogen effects are mediated by estrogen receptor ? (ER?), which reduces chromatin occupancy and transcriptional activity of double homeobox 4 (DUX4), a protein whose aberrant expression has been implicated in FSHD pathogenesis. During myoblast differentiation, we observed that the levels and activity of DUX4 increased progressively and were associated with its enhanced recruitment in the nucleus. ER? interfered with this recruitment by relocalizing DUX4 in the cytoplasm. This work identifies estrogens as a potential disease modifier that underlie sex-related differences in FSHD by protecting against myoblast differentiation impairments in this disease.",
keywords = "double homeobox 4 protein, estrogen, estrogen receptor beta, homeodomain protein, unclassified drug, DUX4 protein, human, ESR2 protein, human, estradiol, Article, case report, cell differentiation, cell nucleus, cell proliferation, cell survival, cellular distribution, chromatin, controlled study, cytoplasm, facioscapulohumeral muscular dystrophy, female, human, human cell, male, myoblast, priority journal, protein expression, cell culture, gene expression, metabolism, pathology, physiology, protein transport, transcription initiation, Cell Differentiation, Cells, Cultured, Estradiol, Estrogen Receptor beta, Estrogens, Gene Expression, Homeodomain Proteins, Humans, Muscular Dystrophy, Facioscapulohumeral, Myoblasts, Protein Transport, Transcriptional Activation",
author = "E. Teveroni and M. Pellegrino and S. Sacconi and P. Calandra and I. Cascino and S. Farioli-Vecchioli and A. Puma and M. Garibaldi and R. Morosetti and G. Tasca and E. Ricci and C.P. Trevisan and G. Galluzzi and A. Pontecorvi and M. Crescenzi and G. Deidda and F. Moretti",
note = "Export Date: 6 April 2018 CODEN: JCINA Correspondence Address: Moretti, F.; Institute of Cell Biology and Neurobiology, National Research Council of Italy (CNR), Via del Fosso di Fiorano, 64, Italy; email: fabiola.moretti@cnr.it Chemicals/CAS: estradiol, 50-28-2; DUX4 protein, human; ESR2 protein, human; Estradiol; Estrogen Receptor beta; Estrogens; Homeodomain Proteins References: Tawil, R., Van Der Maarel, S.M., Facioscapulohumeral muscular dystrophy (2006) Muscle Nerve, 34 (1), pp. 1-15; Statland, J.M., Donlin-Smith, C.M., Tapscott, S.J., Lemmers, R.J., Van Der Maarel, S.M., Tawil, R., Milder phenotype in facioscapulohumeral dystrophy with 7-10 residual D4Z4 repeats (2015) Neurology, 85 (24), pp. 2147-2150; Zatz, M., Marie, S.K., Cerqueira, A., Vainzof, M., Pavanello, R.C., Passos-Bueno, M.R., The facioscapulohumeral muscular dystrophy (FSHD1) gene affects males more severely and more frequently than females (1998) Am J Med Genet, 77 (2), pp. 155-161; Ricci, E., Progress in the molecular diagnosis of facioscapulohumeral muscular dystrophy and correlation between the number of KpnI repeats at the 4q35 locus and clinical phenotype (1999) Ann Neurol, 45 (6), pp. 751-757; Tonini, M.M., Homozygosity for autosomal dominant facioscapulohumeral muscular dystrophy (FSHD) does not result in a more severe phenotype (2004) J Med Genet, 41 (2), p. e17; Lin, F., Wang, Z.Q., Lin, M.T., Murong, S.X., Wang, N., New insights into genotype-phenotype correlations in Chinese facioscapulohumeral muscular dystrophy: A retrospective analysis of 178 patients (2015) Chin Med J, 128 (13), pp. 1707-1713; Klinge, L., Eagle, M., Haggerty, I.D., Roberts, C.E., Straub, V., Bushby, K.M., Severe phenotype in infantile facioscapulohumeral muscular dystrophy (2006) Neuromuscul Disord, 16 (9-10), pp. 553-558; Lunt, P.W., Correlation between fragment size at D4F104S1 and age at onset or at wheelchair use, with a possible generational effect, accounts for much phenotypic variation in 4q35-facioscapulohumeral muscular dystrophy (FSHD) (1995) Hum Mol Genet, 4 (5), pp. 951-958; Rudnik-Sch{\"o}neborn, S., Glauner, B., R{\"o}hrig, D., Zerres, K., Obstetric aspects in women with facioscapulohumeral muscular dystrophy, limb-girdle muscular dystrophy, and congenital myopathies (1997) Arch Neurol, 54 (7), pp. 888-894; Awater, C., Zerres, K., Rudnik-Sch{\"o}neborn, S., Pregnancy course and outcome in women with hereditary neuromuscular disorders: Comparison of obstetric risks in 178 patients (2012) Eur J Obstet Gynecol Reprod Biol, 162 (2), pp. 153-159; Ciafaloni, E., Pregnancy and birth outcomes in women with facioscapulohumeral muscular dystrophy (2006) Neurology, 67 (10), pp. 1887-1889; Sacconi, S., Salviati, L., Desnuelle, C., Facioscapulohumeral muscular dystrophy (2015) Biochim Biophys Acta, 1852 (4), pp. 607-614; Tawil, R., Van Der Maarel, S.M., Tapscott, S.J., Facioscapulohumeral dystrophy: The path to consensus on pathophysiology (2014) Skelet Muscle, 4, p. 12; Calandra, P., Allele-specific DNA hypomethylation characterises FSHD1 and FSHD2 (2016) J Med Genet, 53 (5), pp. 348-355; Van Overveld, P.G., Hypomethylation of D4Z4 in 4q-linked and non-4q-linked facioscapulohumeral muscular dystrophy (2003) Nat Genet, 35 (4), pp. 315-317; Lemmers, R.J., Inter-individual differences in CpG methylation at D4Z4 correlate with clinical variability in FSHD1 and FSHD2 (2015) Hum Mol Genet, 24 (3), pp. 659-669; Geng, L.N., DUX4 activates germline genes, retroelements, and immune mediators: Implications for facioscapulohumeral dystrophy (2012) Dev Cell, 22 (1), pp. 38-51; Daxinger, L., Tapscott, S.J., Van Der Maarel, S.M., Genetic and epigenetic contributors to FSHD (2015) Curr Opin Genet Dev, 33, pp. 56-61; Dieli-Conwright, C.M., Spektor, T.M., Rice, J.C., Todd Schroeder, E., Oestradiol and SERM treatments influence oestrogen receptor coregulator gene expression in human skeletal muscle cells (2009) Acta Physiol (Oxf), 197 (3), pp. 187-196; Wiik, A., Expression of oestrogen receptor ? and ? is higher in skeletal muscle of highly endurance-trained than of moderately active men (2005) Acta Physiol Scand, 184 (2), pp. 105-112; Wiik, A., Ekman, M., Johansson, O., Jansson, E., Esbj{\"o}rnsson, M., Expression of both oestrogen receptor ? and ? in human skeletal muscle tissue (2009) Histochem Cell Biol, 131 (2), pp. 181-189; Wiik, A., Oestrogen receptor ? is expressed in adult human skeletal muscle both at the mRNA and protein level (2003) Acta Physiol Scand, 179 (4), pp. 381-387; Shiomi, K., CDK4 and cyclin D1 allow human myogenic cells to recapture growth property without compromising differentiation potential (2011) Gene Ther, 18 (9), pp. 857-866; Barro, M., Carnac, G., Flavier, S., Mercier, J., Vassetzky, Y., Laoudj-Chenivesse, D., Myoblasts from affected and non-affected FSHD muscles exhibit morphological differentiation defects (2010) J Cell Mol Med, 14 (1-2), pp. 275-289; Tassin, A., DUX4 expression in FSHD muscle cells: How could such a rare protein cause a myopathy? (2013) J Cell Mol Med, 17 (1), pp. 76-89; Dmitriev, P., DUX4-induced constitutive DNA damage and oxidative stress contribute to aberrant differentiation of myoblasts from FSHD patients (2016) Free Radic Biol Med, 99, pp. 244-258; Knopp, P., DUX4 induces a transcriptome more characteristic of a less-differentiated cell state and inhibits myogenesis (2016) J Cell Sci, 129 (20), pp. 3816-3831; Yakimchuk, K., Jondal, M., Okret, S., Estrogen receptor ? and ? in the normal immune system and in lymphoid malignancies (2013) Mol Cell Endocrinol, 375 (1-2), pp. 121-129; Thomas, C., Gustafsson, J.{\AA}., The different roles of ER subtypes in cancer biology and therapy (2011) Nat Rev Cancer, 11 (8), pp. 597-608; Lemmers, R.J., A unifying genetic model for facioscapulohumeral muscular dystrophy (2010) Science, 329 (5999), pp. 1650-1653; Wallace, L.M., DUX4, a candidate gene for facioscapulohumeral muscular dystrophy, causes p53-dependent myopathy in vivo (2011) Ann Neurol, 69 (3), pp. 540-552; Bosnakovski, D., An isogenetic myoblast expression screen identifies DUX4-mediated FSHD-associated molecular pathologies (2008) EMBO J, 27 (20), pp. 2766-2779; Vanderplanck, C., The FSHD atrophic myotube phenotype is caused by DUX4 expression (2011) PLoS One, 6 (10), p. e26820; Bosnakovski, D., Daughters, R.S., Xu, Z., Slack, J.M., Kyba, M., Biphasic myopathic phenotype of mouse DUX, an ORF within conserved FSHD-related repeats (2009) PLoS One, 4 (9), p. e7003; Sappok, A., Mahlknecht, U., Ribavirin restores ESR1 gene expression and tamoxifen sensitivity in ESR1 negative breast cancer cell lines (2011) Clin Epigenetics, 3, p. 8; O'Donnell, A.J., Macleod, K.G., Burns, D.J., Smyth, J.F., Langdon, S.P., Estrogen receptor-alpha mediates gene expression changes and growth response in ovarian cancer cells exposed to estrogen (2005) Endocr Relat Cancer, 12 (4), pp. 851-866; Wijayaratne, A.L., McDonnell, D.P., The human estrogen receptor-alpha is a ubiquitinated protein whose stability is affected differentially by agonists, antagonists, and selective estrogen receptor modulators (2001) J Biol Chem, 276 (38), pp. 35684-35692; Meyers, M.J., Sun, J., Carlson, K.E., Marriner, G.A., Katzenellenbogen, B.S., Katzenellenbogen, J.A., Estrogen receptor-beta potency-selective ligands: Structure-activity relationship studies of diarylpropionitriles and their acetylene and polar analogues (2001) J Med Chem, 44 (24), pp. 4230-4251; Ferreboeuf, M., DUX4 and DUX4 downstream target genes are expressed in fetal FSHD muscles (2014) Hum Mol Genet, 23 (1), pp. 171-181; Rickard, A.M., Petek, L.M., Miller, D.G., Endogenous DUX4 expression in FSHD myotubes is sufficient to cause cell death and disrupts RNA splicing and cell migration pathways (2015) Hum Mol Genet, 24 (20), pp. 5901-5914; Zhang, Y., DNA-binding sequence specificity of DUX4 (2016) Skelet Muscle, 6, p. 8; Dixit, M., DUX4, a candidate gene of facioscapulohumeral muscular dystrophy, encodes a transcriptional activator of PITX1 (2007) Proc Natl Acad Sci U S A, 104 (46), pp. 18157-18162; Pandey, S.N., Conditional over-expression of PITX1 causes skeletal muscle dystrophy in mice (2012) Biol Open, 1 (7), pp. 629-639; Klooster, R., Comprehensive expression analysis of FSHD candidate genes at the mRNA and protein level (2009) Eur J Hum Genet, 17 (12), pp. 1615-1624; Pinto, G., Systematic nucleo-cytoplasmic trafficking of proteins following exposure of MCF7 breast cancer cells to estradiol (2014) J Proteome Res, 13 (2), pp. 1112-1127; Corona, E.D., Jacquelin, D., Gatica, L., Rosa, A.L., Multiple protein domains contribute to nuclear import and cell toxicity of DUX4, a candidate pathogenic protein for facioscapulohumeral muscular dystrophy (2013) PLoS One, 8 (10), p. e75614; Tawil, R., Evidence-based guideline summary: Evaluation, diagnosis, and management of facioscapulohumeral muscular dystrophy: Report of the Guideline Development, Dissemination, and Implementation Subcommittee of the American Academy of Neurology and the Practice Issues Review Panel of the American Association of Neuromuscular & Electrodiagnostic Medicine (2015) Neurology, 85 (4), pp. 357-364; Broccolini, A., Neprilysin participates in skeletal muscle regeneration and is accumulated in abnormal muscle fibres of inclusion body myositis (2006) J Neurochem, 96 (3), pp. 777-789; Bader, D., Masaki, T., Fischman, D.A., Immunochemical analysis of myosin heavy chain during avian myogenesis in vivo and in vitro (1982) J Cell Biol, 95 (3), pp. 763-770; Balog, J., Increased DUX4 expression during muscle differentiation correlates with decreased SMCHD1 protein levels at D4Z4 (2015) Epigenetics, 10 (12), pp. 1133-1142; Pfaffl, M.W., A new mathematical model for relative quantification in real-time RT-PCR (2001) Nucleic Acids Res, 29 (9), p. e45; Chen, J., Marechal, V., Levine, A.J., Mapping of the p53 and mdm-2 interaction domains (1993) Mol Cell Biol, 13 (7), pp. 4107-4114; Consalvi, S., Brancaccio, A., Dall'Agnese, A., Puri, P.L., Palacios, D., Praja1 E3 ubiquitin ligase promotes skeletal myogenesis through degradation of EZH2 upon p38? activation (2017) Nat Commun, 8, p. 13956; Pellegrino, M., Targeting the MDM2/MDM4 interaction interface as a promising approach for p53 reactivation therapy (2015) Cancer Res, 75 (21), pp. 4560-4572",
year = "2017",
doi = "10.1172/JCI89401",
language = "English",
volume = "127",
pages = "1531--1545",
journal = "Journal of Clinical Investigation",
issn = "0021-9738",
publisher = "The American Society for Clinical Investigation",
number = "4",

}

TY - JOUR

T1 - Estrogens enhance myoblast differentiation in facioscapulohumeral muscular dystrophy by antagonizing DUX4 activity

AU - Teveroni, E.

AU - Pellegrino, M.

AU - Sacconi, S.

AU - Calandra, P.

AU - Cascino, I.

AU - Farioli-Vecchioli, S.

AU - Puma, A.

AU - Garibaldi, M.

AU - Morosetti, R.

AU - Tasca, G.

AU - Ricci, E.

AU - Trevisan, C.P.

AU - Galluzzi, G.

AU - Pontecorvi, A.

AU - Crescenzi, M.

AU - Deidda, G.

AU - Moretti, F.

N1 - Export Date: 6 April 2018 CODEN: JCINA Correspondence Address: Moretti, F.; Institute of Cell Biology and Neurobiology, National Research Council of Italy (CNR), Via del Fosso di Fiorano, 64, Italy; email: fabiola.moretti@cnr.it Chemicals/CAS: estradiol, 50-28-2; DUX4 protein, human; ESR2 protein, human; Estradiol; Estrogen Receptor beta; Estrogens; Homeodomain Proteins References: Tawil, R., Van Der Maarel, S.M., Facioscapulohumeral muscular dystrophy (2006) Muscle Nerve, 34 (1), pp. 1-15; Statland, J.M., Donlin-Smith, C.M., Tapscott, S.J., Lemmers, R.J., Van Der Maarel, S.M., Tawil, R., Milder phenotype in facioscapulohumeral dystrophy with 7-10 residual D4Z4 repeats (2015) Neurology, 85 (24), pp. 2147-2150; Zatz, M., Marie, S.K., Cerqueira, A., Vainzof, M., Pavanello, R.C., Passos-Bueno, M.R., The facioscapulohumeral muscular dystrophy (FSHD1) gene affects males more severely and more frequently than females (1998) Am J Med Genet, 77 (2), pp. 155-161; Ricci, E., Progress in the molecular diagnosis of facioscapulohumeral muscular dystrophy and correlation between the number of KpnI repeats at the 4q35 locus and clinical phenotype (1999) Ann Neurol, 45 (6), pp. 751-757; Tonini, M.M., Homozygosity for autosomal dominant facioscapulohumeral muscular dystrophy (FSHD) does not result in a more severe phenotype (2004) J Med Genet, 41 (2), p. e17; Lin, F., Wang, Z.Q., Lin, M.T., Murong, S.X., Wang, N., New insights into genotype-phenotype correlations in Chinese facioscapulohumeral muscular dystrophy: A retrospective analysis of 178 patients (2015) Chin Med J, 128 (13), pp. 1707-1713; Klinge, L., Eagle, M., Haggerty, I.D., Roberts, C.E., Straub, V., Bushby, K.M., Severe phenotype in infantile facioscapulohumeral muscular dystrophy (2006) Neuromuscul Disord, 16 (9-10), pp. 553-558; Lunt, P.W., Correlation between fragment size at D4F104S1 and age at onset or at wheelchair use, with a possible generational effect, accounts for much phenotypic variation in 4q35-facioscapulohumeral muscular dystrophy (FSHD) (1995) Hum Mol Genet, 4 (5), pp. 951-958; Rudnik-Schöneborn, S., Glauner, B., Röhrig, D., Zerres, K., Obstetric aspects in women with facioscapulohumeral muscular dystrophy, limb-girdle muscular dystrophy, and congenital myopathies (1997) Arch Neurol, 54 (7), pp. 888-894; Awater, C., Zerres, K., Rudnik-Schöneborn, S., Pregnancy course and outcome in women with hereditary neuromuscular disorders: Comparison of obstetric risks in 178 patients (2012) Eur J Obstet Gynecol Reprod Biol, 162 (2), pp. 153-159; Ciafaloni, E., Pregnancy and birth outcomes in women with facioscapulohumeral muscular dystrophy (2006) Neurology, 67 (10), pp. 1887-1889; Sacconi, S., Salviati, L., Desnuelle, C., Facioscapulohumeral muscular dystrophy (2015) Biochim Biophys Acta, 1852 (4), pp. 607-614; Tawil, R., Van Der Maarel, S.M., Tapscott, S.J., Facioscapulohumeral dystrophy: The path to consensus on pathophysiology (2014) Skelet Muscle, 4, p. 12; Calandra, P., Allele-specific DNA hypomethylation characterises FSHD1 and FSHD2 (2016) J Med Genet, 53 (5), pp. 348-355; Van Overveld, P.G., Hypomethylation of D4Z4 in 4q-linked and non-4q-linked facioscapulohumeral muscular dystrophy (2003) Nat Genet, 35 (4), pp. 315-317; Lemmers, R.J., Inter-individual differences in CpG methylation at D4Z4 correlate with clinical variability in FSHD1 and FSHD2 (2015) Hum Mol Genet, 24 (3), pp. 659-669; Geng, L.N., DUX4 activates germline genes, retroelements, and immune mediators: Implications for facioscapulohumeral dystrophy (2012) Dev Cell, 22 (1), pp. 38-51; Daxinger, L., Tapscott, S.J., Van Der Maarel, S.M., Genetic and epigenetic contributors to FSHD (2015) Curr Opin Genet Dev, 33, pp. 56-61; Dieli-Conwright, C.M., Spektor, T.M., Rice, J.C., Todd Schroeder, E., Oestradiol and SERM treatments influence oestrogen receptor coregulator gene expression in human skeletal muscle cells (2009) Acta Physiol (Oxf), 197 (3), pp. 187-196; Wiik, A., Expression of oestrogen receptor ? and ? is higher in skeletal muscle of highly endurance-trained than of moderately active men (2005) Acta Physiol Scand, 184 (2), pp. 105-112; Wiik, A., Ekman, M., Johansson, O., Jansson, E., Esbjörnsson, M., Expression of both oestrogen receptor ? and ? in human skeletal muscle tissue (2009) Histochem Cell Biol, 131 (2), pp. 181-189; Wiik, A., Oestrogen receptor ? is expressed in adult human skeletal muscle both at the mRNA and protein level (2003) Acta Physiol Scand, 179 (4), pp. 381-387; Shiomi, K., CDK4 and cyclin D1 allow human myogenic cells to recapture growth property without compromising differentiation potential (2011) Gene Ther, 18 (9), pp. 857-866; Barro, M., Carnac, G., Flavier, S., Mercier, J., Vassetzky, Y., Laoudj-Chenivesse, D., Myoblasts from affected and non-affected FSHD muscles exhibit morphological differentiation defects (2010) J Cell Mol Med, 14 (1-2), pp. 275-289; Tassin, A., DUX4 expression in FSHD muscle cells: How could such a rare protein cause a myopathy? (2013) J Cell Mol Med, 17 (1), pp. 76-89; Dmitriev, P., DUX4-induced constitutive DNA damage and oxidative stress contribute to aberrant differentiation of myoblasts from FSHD patients (2016) Free Radic Biol Med, 99, pp. 244-258; Knopp, P., DUX4 induces a transcriptome more characteristic of a less-differentiated cell state and inhibits myogenesis (2016) J Cell Sci, 129 (20), pp. 3816-3831; Yakimchuk, K., Jondal, M., Okret, S., Estrogen receptor ? and ? in the normal immune system and in lymphoid malignancies (2013) Mol Cell Endocrinol, 375 (1-2), pp. 121-129; Thomas, C., Gustafsson, J.Å., The different roles of ER subtypes in cancer biology and therapy (2011) Nat Rev Cancer, 11 (8), pp. 597-608; Lemmers, R.J., A unifying genetic model for facioscapulohumeral muscular dystrophy (2010) Science, 329 (5999), pp. 1650-1653; Wallace, L.M., DUX4, a candidate gene for facioscapulohumeral muscular dystrophy, causes p53-dependent myopathy in vivo (2011) Ann Neurol, 69 (3), pp. 540-552; Bosnakovski, D., An isogenetic myoblast expression screen identifies DUX4-mediated FSHD-associated molecular pathologies (2008) EMBO J, 27 (20), pp. 2766-2779; Vanderplanck, C., The FSHD atrophic myotube phenotype is caused by DUX4 expression (2011) PLoS One, 6 (10), p. e26820; Bosnakovski, D., Daughters, R.S., Xu, Z., Slack, J.M., Kyba, M., Biphasic myopathic phenotype of mouse DUX, an ORF within conserved FSHD-related repeats (2009) PLoS One, 4 (9), p. e7003; Sappok, A., Mahlknecht, U., Ribavirin restores ESR1 gene expression and tamoxifen sensitivity in ESR1 negative breast cancer cell lines (2011) Clin Epigenetics, 3, p. 8; O'Donnell, A.J., Macleod, K.G., Burns, D.J., Smyth, J.F., Langdon, S.P., Estrogen receptor-alpha mediates gene expression changes and growth response in ovarian cancer cells exposed to estrogen (2005) Endocr Relat Cancer, 12 (4), pp. 851-866; Wijayaratne, A.L., McDonnell, D.P., The human estrogen receptor-alpha is a ubiquitinated protein whose stability is affected differentially by agonists, antagonists, and selective estrogen receptor modulators (2001) J Biol Chem, 276 (38), pp. 35684-35692; Meyers, M.J., Sun, J., Carlson, K.E., Marriner, G.A., Katzenellenbogen, B.S., Katzenellenbogen, J.A., Estrogen receptor-beta potency-selective ligands: Structure-activity relationship studies of diarylpropionitriles and their acetylene and polar analogues (2001) J Med Chem, 44 (24), pp. 4230-4251; Ferreboeuf, M., DUX4 and DUX4 downstream target genes are expressed in fetal FSHD muscles (2014) Hum Mol Genet, 23 (1), pp. 171-181; Rickard, A.M., Petek, L.M., Miller, D.G., Endogenous DUX4 expression in FSHD myotubes is sufficient to cause cell death and disrupts RNA splicing and cell migration pathways (2015) Hum Mol Genet, 24 (20), pp. 5901-5914; Zhang, Y., DNA-binding sequence specificity of DUX4 (2016) Skelet Muscle, 6, p. 8; Dixit, M., DUX4, a candidate gene of facioscapulohumeral muscular dystrophy, encodes a transcriptional activator of PITX1 (2007) Proc Natl Acad Sci U S A, 104 (46), pp. 18157-18162; Pandey, S.N., Conditional over-expression of PITX1 causes skeletal muscle dystrophy in mice (2012) Biol Open, 1 (7), pp. 629-639; Klooster, R., Comprehensive expression analysis of FSHD candidate genes at the mRNA and protein level (2009) Eur J Hum Genet, 17 (12), pp. 1615-1624; Pinto, G., Systematic nucleo-cytoplasmic trafficking of proteins following exposure of MCF7 breast cancer cells to estradiol (2014) J Proteome Res, 13 (2), pp. 1112-1127; Corona, E.D., Jacquelin, D., Gatica, L., Rosa, A.L., Multiple protein domains contribute to nuclear import and cell toxicity of DUX4, a candidate pathogenic protein for facioscapulohumeral muscular dystrophy (2013) PLoS One, 8 (10), p. e75614; Tawil, R., Evidence-based guideline summary: Evaluation, diagnosis, and management of facioscapulohumeral muscular dystrophy: Report of the Guideline Development, Dissemination, and Implementation Subcommittee of the American Academy of Neurology and the Practice Issues Review Panel of the American Association of Neuromuscular & Electrodiagnostic Medicine (2015) Neurology, 85 (4), pp. 357-364; Broccolini, A., Neprilysin participates in skeletal muscle regeneration and is accumulated in abnormal muscle fibres of inclusion body myositis (2006) J Neurochem, 96 (3), pp. 777-789; Bader, D., Masaki, T., Fischman, D.A., Immunochemical analysis of myosin heavy chain during avian myogenesis in vivo and in vitro (1982) J Cell Biol, 95 (3), pp. 763-770; Balog, J., Increased DUX4 expression during muscle differentiation correlates with decreased SMCHD1 protein levels at D4Z4 (2015) Epigenetics, 10 (12), pp. 1133-1142; Pfaffl, M.W., A new mathematical model for relative quantification in real-time RT-PCR (2001) Nucleic Acids Res, 29 (9), p. e45; Chen, J., Marechal, V., Levine, A.J., Mapping of the p53 and mdm-2 interaction domains (1993) Mol Cell Biol, 13 (7), pp. 4107-4114; Consalvi, S., Brancaccio, A., Dall'Agnese, A., Puri, P.L., Palacios, D., Praja1 E3 ubiquitin ligase promotes skeletal myogenesis through degradation of EZH2 upon p38? activation (2017) Nat Commun, 8, p. 13956; Pellegrino, M., Targeting the MDM2/MDM4 interaction interface as a promising approach for p53 reactivation therapy (2015) Cancer Res, 75 (21), pp. 4560-4572

PY - 2017

Y1 - 2017

N2 - Facioscapulohumeral muscular dystrophy (FSHD) is an autosomal dominant neuromuscular disorder that is characterized by extreme variability in symptoms, with females being less severely affected than males and presenting a higher proportion of asymptomatic carriers. The sex-related factors involved in the disease are not known. Here, we have utilized myoblasts isolated from FSHD patients (FSHD myoblasts) to investigate the effect of estrogens on muscle properties. Our results demonstrated that estrogens counteract the differentiation impairment of FSHD myoblasts without affecting cell proliferation or survival. Estrogen effects are mediated by estrogen receptor ? (ER?), which reduces chromatin occupancy and transcriptional activity of double homeobox 4 (DUX4), a protein whose aberrant expression has been implicated in FSHD pathogenesis. During myoblast differentiation, we observed that the levels and activity of DUX4 increased progressively and were associated with its enhanced recruitment in the nucleus. ER? interfered with this recruitment by relocalizing DUX4 in the cytoplasm. This work identifies estrogens as a potential disease modifier that underlie sex-related differences in FSHD by protecting against myoblast differentiation impairments in this disease.

AB - Facioscapulohumeral muscular dystrophy (FSHD) is an autosomal dominant neuromuscular disorder that is characterized by extreme variability in symptoms, with females being less severely affected than males and presenting a higher proportion of asymptomatic carriers. The sex-related factors involved in the disease are not known. Here, we have utilized myoblasts isolated from FSHD patients (FSHD myoblasts) to investigate the effect of estrogens on muscle properties. Our results demonstrated that estrogens counteract the differentiation impairment of FSHD myoblasts without affecting cell proliferation or survival. Estrogen effects are mediated by estrogen receptor ? (ER?), which reduces chromatin occupancy and transcriptional activity of double homeobox 4 (DUX4), a protein whose aberrant expression has been implicated in FSHD pathogenesis. During myoblast differentiation, we observed that the levels and activity of DUX4 increased progressively and were associated with its enhanced recruitment in the nucleus. ER? interfered with this recruitment by relocalizing DUX4 in the cytoplasm. This work identifies estrogens as a potential disease modifier that underlie sex-related differences in FSHD by protecting against myoblast differentiation impairments in this disease.

KW - double homeobox 4 protein

KW - estrogen

KW - estrogen receptor beta

KW - homeodomain protein

KW - unclassified drug

KW - DUX4 protein, human

KW - ESR2 protein, human

KW - estradiol

KW - Article

KW - case report

KW - cell differentiation

KW - cell nucleus

KW - cell proliferation

KW - cell survival

KW - cellular distribution

KW - chromatin

KW - controlled study

KW - cytoplasm

KW - facioscapulohumeral muscular dystrophy

KW - female

KW - human

KW - human cell

KW - male

KW - myoblast

KW - priority journal

KW - protein expression

KW - cell culture

KW - gene expression

KW - metabolism

KW - pathology

KW - physiology

KW - protein transport

KW - transcription initiation

KW - Cell Differentiation

KW - Cells, Cultured

KW - Estradiol

KW - Estrogen Receptor beta

KW - Estrogens

KW - Gene Expression

KW - Homeodomain Proteins

KW - Humans

KW - Muscular Dystrophy, Facioscapulohumeral

KW - Myoblasts

KW - Protein Transport

KW - Transcriptional Activation

U2 - 10.1172/JCI89401

DO - 10.1172/JCI89401

M3 - Article

VL - 127

SP - 1531

EP - 1545

JO - Journal of Clinical Investigation

JF - Journal of Clinical Investigation

SN - 0021-9738

IS - 4

ER -