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Consequences of Lmna Exon 4 Mutations in Myoblast Function

Laminopathies are causally associated with mutations on the Lamin A/C gene (LMNA). To date, more than 400 mutations in LMNA have been reported in patients. These mutations are widely distributed throughout the entire gene and are associated with a wide range of phenotypes. Unfortunately, little is k...

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Autores principales: Gómez-Domínguez, Déborah, Epifano, Carolina, de Miguel, Fernando, Castaño, Albert García, Vilaplana-Martí, Borja, Martín, Alberto, Amarilla-Quintana, Sandra, Bertrand, Anne T, Bonne, Gisèle, Ramón-Azcón, Javier, Rodríguez-Milla, Miguel A, Pérez de Castro, Ignacio
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7291140/
https://www.ncbi.nlm.nih.gov/pubmed/32455813
http://dx.doi.org/10.3390/cells9051286
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author Gómez-Domínguez, Déborah
Epifano, Carolina
de Miguel, Fernando
Castaño, Albert García
Vilaplana-Martí, Borja
Martín, Alberto
Amarilla-Quintana, Sandra
Bertrand, Anne T
Bonne, Gisèle
Ramón-Azcón, Javier
Rodríguez-Milla, Miguel A
Pérez de Castro, Ignacio
author_facet Gómez-Domínguez, Déborah
Epifano, Carolina
de Miguel, Fernando
Castaño, Albert García
Vilaplana-Martí, Borja
Martín, Alberto
Amarilla-Quintana, Sandra
Bertrand, Anne T
Bonne, Gisèle
Ramón-Azcón, Javier
Rodríguez-Milla, Miguel A
Pérez de Castro, Ignacio
author_sort Gómez-Domínguez, Déborah
collection PubMed
description Laminopathies are causally associated with mutations on the Lamin A/C gene (LMNA). To date, more than 400 mutations in LMNA have been reported in patients. These mutations are widely distributed throughout the entire gene and are associated with a wide range of phenotypes. Unfortunately, little is known about the mechanisms underlying the effect of the majority of these mutations. This is the case of more than 40 mutations that are located at exon 4. Using CRISPR/Cas9 technology, we generated a collection of Lmna exon 4 mutants in mouse C2C12 myoblasts. These cell models included different types of exon 4 deletions and the presence of R249W mutation, one of the human variants associated with a severe type of laminopathy, LMNA-associated congenital muscular dystrophy (L-CMD). We characterized these clones by measuring their nuclear circularity, myogenic differentiation capacity in 2D and 3D conditions, DNA damage, and levels of p-ERK and p-AKT (phosphorylated Mitogen-Activated Protein Kinase 1/3 and AKT serine/threonine kinase 1). Our results indicated that Lmna exon 4 mutants showed abnormal nuclear morphology. In addition, levels and/or subcellular localization of different members of the lamin and LINC (LInker of Nucleoskeleton and Cytoskeleton) complex were altered in all these mutants. Whereas no significant differences were observed for ERK and AKT activities, the accumulation of DNA damage was associated to the Lmna p.R249W mutant myoblasts. Finally, significant myogenic differentiation defects were detected in the Lmna exon 4 mutants. These results have key implications in the development of future therapeutic strategies for the treatment of laminopathies.
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spelling pubmed-72911402020-06-17 Consequences of Lmna Exon 4 Mutations in Myoblast Function Gómez-Domínguez, Déborah Epifano, Carolina de Miguel, Fernando Castaño, Albert García Vilaplana-Martí, Borja Martín, Alberto Amarilla-Quintana, Sandra Bertrand, Anne T Bonne, Gisèle Ramón-Azcón, Javier Rodríguez-Milla, Miguel A Pérez de Castro, Ignacio Cells Article Laminopathies are causally associated with mutations on the Lamin A/C gene (LMNA). To date, more than 400 mutations in LMNA have been reported in patients. These mutations are widely distributed throughout the entire gene and are associated with a wide range of phenotypes. Unfortunately, little is known about the mechanisms underlying the effect of the majority of these mutations. This is the case of more than 40 mutations that are located at exon 4. Using CRISPR/Cas9 technology, we generated a collection of Lmna exon 4 mutants in mouse C2C12 myoblasts. These cell models included different types of exon 4 deletions and the presence of R249W mutation, one of the human variants associated with a severe type of laminopathy, LMNA-associated congenital muscular dystrophy (L-CMD). We characterized these clones by measuring their nuclear circularity, myogenic differentiation capacity in 2D and 3D conditions, DNA damage, and levels of p-ERK and p-AKT (phosphorylated Mitogen-Activated Protein Kinase 1/3 and AKT serine/threonine kinase 1). Our results indicated that Lmna exon 4 mutants showed abnormal nuclear morphology. In addition, levels and/or subcellular localization of different members of the lamin and LINC (LInker of Nucleoskeleton and Cytoskeleton) complex were altered in all these mutants. Whereas no significant differences were observed for ERK and AKT activities, the accumulation of DNA damage was associated to the Lmna p.R249W mutant myoblasts. Finally, significant myogenic differentiation defects were detected in the Lmna exon 4 mutants. These results have key implications in the development of future therapeutic strategies for the treatment of laminopathies. MDPI 2020-05-21 /pmc/articles/PMC7291140/ /pubmed/32455813 http://dx.doi.org/10.3390/cells9051286 Text en © 2020 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Gómez-Domínguez, Déborah
Epifano, Carolina
de Miguel, Fernando
Castaño, Albert García
Vilaplana-Martí, Borja
Martín, Alberto
Amarilla-Quintana, Sandra
Bertrand, Anne T
Bonne, Gisèle
Ramón-Azcón, Javier
Rodríguez-Milla, Miguel A
Pérez de Castro, Ignacio
Consequences of Lmna Exon 4 Mutations in Myoblast Function
title Consequences of Lmna Exon 4 Mutations in Myoblast Function
title_full Consequences of Lmna Exon 4 Mutations in Myoblast Function
title_fullStr Consequences of Lmna Exon 4 Mutations in Myoblast Function
title_full_unstemmed Consequences of Lmna Exon 4 Mutations in Myoblast Function
title_short Consequences of Lmna Exon 4 Mutations in Myoblast Function
title_sort consequences of lmna exon 4 mutations in myoblast function
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7291140/
https://www.ncbi.nlm.nih.gov/pubmed/32455813
http://dx.doi.org/10.3390/cells9051286
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