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Dnmt3b regulates DUX4 expression in a tissue-dependent manner in transgenic D4Z4 mice
BACKGROUND: Facioscapulohumeral muscular dystrophy (FSHD) is a skeletal muscle disorder that is caused by derepression of the transcription factor DUX4 in skeletal muscle cells. Apart from SMCHD1, DNMT3B was recently identified as a disease gene and disease modifier in FSHD. However, the exact role...
Autores principales: | , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
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BioMed Central
2020
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7528343/ https://www.ncbi.nlm.nih.gov/pubmed/33004076 http://dx.doi.org/10.1186/s13395-020-00247-0 |
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author | Bouwman, Linde F. den Hamer, Bianca Verveer, Elwin P. Lerink, Lente J. S. Krom, Yvonne D. van der Maarel, Silvère M. de Greef, Jessica C. |
author_facet | Bouwman, Linde F. den Hamer, Bianca Verveer, Elwin P. Lerink, Lente J. S. Krom, Yvonne D. van der Maarel, Silvère M. de Greef, Jessica C. |
author_sort | Bouwman, Linde F. |
collection | PubMed |
description | BACKGROUND: Facioscapulohumeral muscular dystrophy (FSHD) is a skeletal muscle disorder that is caused by derepression of the transcription factor DUX4 in skeletal muscle cells. Apart from SMCHD1, DNMT3B was recently identified as a disease gene and disease modifier in FSHD. However, the exact role of DNMT3B at the D4Z4 repeat array remains unknown. METHODS: To determine the role of Dnmt3b on DUX4 repression, hemizygous mice with a FSHD-sized D4Z4 repeat array (D4Z4-2.5 mice) were cross-bred with mice carrying an in-frame exon skipping mutation in Dnmt3b (Dnmt3b(MommeD14) mice). Additionally, siRNA knockdowns of Dnmt3b were performed in mouse embryonic stem cells (mESCs) derived from the D4Z4-2.5 mouse model. RESULTS: In mESCs derived from D4Z4-2.5 mice, Dnmt3b was enriched at the D4Z4 repeat array and DUX4 transcript levels were upregulated after a knockdown of Dnmt3b. In D4Z4-2.5/Dnmt3b(MommeD14) mice, Dnmt3b protein levels were reduced; however, DUX4 RNA levels in skeletal muscles were not enhanced and no pathology was observed. Interestingly, D4Z4-2.5/Dnmt3b(MommeD14) mice showed a loss of DNA methylation at the D4Z4 repeat array and significantly higher DUX4 transcript levels in secondary lymphoid organs. As these lymphoid organs seem to be more sensitive to epigenetic modifiers of the D4Z4 repeat array, different immune cell populations were quantified in the spleen and inguinal lymph nodes of D4Z4-2.5 mice crossed with Dnmt3b(MommeD14) mice or Smchd1(MommeD1) mice. Only in D4Z4-2.5/Smchd1(MommeD1) mice the immune cell populations were disturbed. CONCLUSIONS: Our data demonstrates that loss of Dnmt3b results in derepression of DUX4 in lymphoid tissues and mESCs but not in myogenic cells of D4Z4-2.5/Dnmt3b(MommeD14) mice. In addition, the Smchd1(MommeD1) variant seems to have a more potent role in DUX4 derepression. Our studies suggest that the immune system is particularly but differentially sensitive to D4Z4 chromatin modifiers which may provide a molecular basis for the yet underexplored immune involvement in FSHD. |
format | Online Article Text |
id | pubmed-7528343 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2020 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-75283432020-10-02 Dnmt3b regulates DUX4 expression in a tissue-dependent manner in transgenic D4Z4 mice Bouwman, Linde F. den Hamer, Bianca Verveer, Elwin P. Lerink, Lente J. S. Krom, Yvonne D. van der Maarel, Silvère M. de Greef, Jessica C. Skelet Muscle Research BACKGROUND: Facioscapulohumeral muscular dystrophy (FSHD) is a skeletal muscle disorder that is caused by derepression of the transcription factor DUX4 in skeletal muscle cells. Apart from SMCHD1, DNMT3B was recently identified as a disease gene and disease modifier in FSHD. However, the exact role of DNMT3B at the D4Z4 repeat array remains unknown. METHODS: To determine the role of Dnmt3b on DUX4 repression, hemizygous mice with a FSHD-sized D4Z4 repeat array (D4Z4-2.5 mice) were cross-bred with mice carrying an in-frame exon skipping mutation in Dnmt3b (Dnmt3b(MommeD14) mice). Additionally, siRNA knockdowns of Dnmt3b were performed in mouse embryonic stem cells (mESCs) derived from the D4Z4-2.5 mouse model. RESULTS: In mESCs derived from D4Z4-2.5 mice, Dnmt3b was enriched at the D4Z4 repeat array and DUX4 transcript levels were upregulated after a knockdown of Dnmt3b. In D4Z4-2.5/Dnmt3b(MommeD14) mice, Dnmt3b protein levels were reduced; however, DUX4 RNA levels in skeletal muscles were not enhanced and no pathology was observed. Interestingly, D4Z4-2.5/Dnmt3b(MommeD14) mice showed a loss of DNA methylation at the D4Z4 repeat array and significantly higher DUX4 transcript levels in secondary lymphoid organs. As these lymphoid organs seem to be more sensitive to epigenetic modifiers of the D4Z4 repeat array, different immune cell populations were quantified in the spleen and inguinal lymph nodes of D4Z4-2.5 mice crossed with Dnmt3b(MommeD14) mice or Smchd1(MommeD1) mice. Only in D4Z4-2.5/Smchd1(MommeD1) mice the immune cell populations were disturbed. CONCLUSIONS: Our data demonstrates that loss of Dnmt3b results in derepression of DUX4 in lymphoid tissues and mESCs but not in myogenic cells of D4Z4-2.5/Dnmt3b(MommeD14) mice. In addition, the Smchd1(MommeD1) variant seems to have a more potent role in DUX4 derepression. Our studies suggest that the immune system is particularly but differentially sensitive to D4Z4 chromatin modifiers which may provide a molecular basis for the yet underexplored immune involvement in FSHD. BioMed Central 2020-10-01 /pmc/articles/PMC7528343/ /pubmed/33004076 http://dx.doi.org/10.1186/s13395-020-00247-0 Text en © The Author(s) 2020 Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data. |
spellingShingle | Research Bouwman, Linde F. den Hamer, Bianca Verveer, Elwin P. Lerink, Lente J. S. Krom, Yvonne D. van der Maarel, Silvère M. de Greef, Jessica C. Dnmt3b regulates DUX4 expression in a tissue-dependent manner in transgenic D4Z4 mice |
title | Dnmt3b regulates DUX4 expression in a tissue-dependent manner in transgenic D4Z4 mice |
title_full | Dnmt3b regulates DUX4 expression in a tissue-dependent manner in transgenic D4Z4 mice |
title_fullStr | Dnmt3b regulates DUX4 expression in a tissue-dependent manner in transgenic D4Z4 mice |
title_full_unstemmed | Dnmt3b regulates DUX4 expression in a tissue-dependent manner in transgenic D4Z4 mice |
title_short | Dnmt3b regulates DUX4 expression in a tissue-dependent manner in transgenic D4Z4 mice |
title_sort | dnmt3b regulates dux4 expression in a tissue-dependent manner in transgenic d4z4 mice |
topic | Research |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7528343/ https://www.ncbi.nlm.nih.gov/pubmed/33004076 http://dx.doi.org/10.1186/s13395-020-00247-0 |
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