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Integrative Cell Type-Specific Multi-Omics Approaches Reveal Impaired Programs of Glial Cell Differentiation in Mouse Culture Models of DM1

Myotonic dystrophy type 1 (DM1) is a neuromuscular disorder caused by a non-coding CTG repeat expansion in the DMPK gene. This mutation generates a toxic CUG RNA that interferes with the RNA processing of target genes in multiple tissues. Despite debilitating neurological impairment, the pathophysio...

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Autores principales: González-Barriga, Anchel, Lallemant, Louison, Dincã, Diana M., Braz, Sandra O., Polvèche, Hélène, Magneron, Paul, Pionneau, Cédric, Huguet-Lachon, Aline, Claude, Jean-Baptiste, Chhuon, Cerina, Guerrera, Ida Chiara, Bourgeois, Cyril F., Auboeuf, Didier, Gourdon, Geneviève, Gomes-Pereira, Mário
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8136287/
https://www.ncbi.nlm.nih.gov/pubmed/34025359
http://dx.doi.org/10.3389/fncel.2021.662035
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author González-Barriga, Anchel
Lallemant, Louison
Dincã, Diana M.
Braz, Sandra O.
Polvèche, Hélène
Magneron, Paul
Pionneau, Cédric
Huguet-Lachon, Aline
Claude, Jean-Baptiste
Chhuon, Cerina
Guerrera, Ida Chiara
Bourgeois, Cyril F.
Auboeuf, Didier
Gourdon, Geneviève
Gomes-Pereira, Mário
author_facet González-Barriga, Anchel
Lallemant, Louison
Dincã, Diana M.
Braz, Sandra O.
Polvèche, Hélène
Magneron, Paul
Pionneau, Cédric
Huguet-Lachon, Aline
Claude, Jean-Baptiste
Chhuon, Cerina
Guerrera, Ida Chiara
Bourgeois, Cyril F.
Auboeuf, Didier
Gourdon, Geneviève
Gomes-Pereira, Mário
author_sort González-Barriga, Anchel
collection PubMed
description Myotonic dystrophy type 1 (DM1) is a neuromuscular disorder caused by a non-coding CTG repeat expansion in the DMPK gene. This mutation generates a toxic CUG RNA that interferes with the RNA processing of target genes in multiple tissues. Despite debilitating neurological impairment, the pathophysiological cascade of molecular and cellular events in the central nervous system (CNS) has been less extensively characterized than the molecular pathogenesis of muscle/cardiac dysfunction. Particularly, the contribution of different cell types to DM1 brain disease is not clearly understood. We first used transcriptomics to compare the impact of expanded CUG RNA on the transcriptome of primary neurons, astrocytes and oligodendrocytes derived from DMSXL mice, a transgenic model of DM1. RNA sequencing revealed more frequent expression and splicing changes in glia than neuronal cells. In particular, primary DMSXL oligodendrocytes showed the highest number of transcripts differentially expressed, while DMSXL astrocytes displayed the most severe splicing dysregulation. Interestingly, the expression and splicing defects of DMSXL glia recreated molecular signatures suggestive of impaired cell differentiation: while DMSXL oligodendrocytes failed to upregulate a subset of genes that are naturally activated during the oligodendroglia differentiation, a significant proportion of missplicing events in DMSXL oligodendrocytes and astrocytes increased the expression of RNA isoforms typical of precursor cell stages. Together these data suggest that expanded CUG RNA in glial cells affects preferentially differentiation-regulated molecular events. This hypothesis was corroborated by gene ontology (GO) analyses, which revealed an enrichment for biological processes and cellular components with critical roles during cell differentiation. Finally, we combined exon ontology with phosphoproteomics and cell imaging to explore the functional impact of CUG-associated spliceopathy on downstream protein metabolism. Changes in phosphorylation, protein isoform expression and intracellular localization in DMSXL astrocytes demonstrate the far-reaching impact of the DM1 repeat expansion on cell metabolism. Our multi-omics approaches provide insight into the mechanisms of CUG RNA toxicity in the CNS with cell type resolution, and support the priority for future research on non-neuronal mechanisms and proteomic changes in DM1 brain disease.
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spelling pubmed-81362872021-05-21 Integrative Cell Type-Specific Multi-Omics Approaches Reveal Impaired Programs of Glial Cell Differentiation in Mouse Culture Models of DM1 González-Barriga, Anchel Lallemant, Louison Dincã, Diana M. Braz, Sandra O. Polvèche, Hélène Magneron, Paul Pionneau, Cédric Huguet-Lachon, Aline Claude, Jean-Baptiste Chhuon, Cerina Guerrera, Ida Chiara Bourgeois, Cyril F. Auboeuf, Didier Gourdon, Geneviève Gomes-Pereira, Mário Front Cell Neurosci Neuroscience Myotonic dystrophy type 1 (DM1) is a neuromuscular disorder caused by a non-coding CTG repeat expansion in the DMPK gene. This mutation generates a toxic CUG RNA that interferes with the RNA processing of target genes in multiple tissues. Despite debilitating neurological impairment, the pathophysiological cascade of molecular and cellular events in the central nervous system (CNS) has been less extensively characterized than the molecular pathogenesis of muscle/cardiac dysfunction. Particularly, the contribution of different cell types to DM1 brain disease is not clearly understood. We first used transcriptomics to compare the impact of expanded CUG RNA on the transcriptome of primary neurons, astrocytes and oligodendrocytes derived from DMSXL mice, a transgenic model of DM1. RNA sequencing revealed more frequent expression and splicing changes in glia than neuronal cells. In particular, primary DMSXL oligodendrocytes showed the highest number of transcripts differentially expressed, while DMSXL astrocytes displayed the most severe splicing dysregulation. Interestingly, the expression and splicing defects of DMSXL glia recreated molecular signatures suggestive of impaired cell differentiation: while DMSXL oligodendrocytes failed to upregulate a subset of genes that are naturally activated during the oligodendroglia differentiation, a significant proportion of missplicing events in DMSXL oligodendrocytes and astrocytes increased the expression of RNA isoforms typical of precursor cell stages. Together these data suggest that expanded CUG RNA in glial cells affects preferentially differentiation-regulated molecular events. This hypothesis was corroborated by gene ontology (GO) analyses, which revealed an enrichment for biological processes and cellular components with critical roles during cell differentiation. Finally, we combined exon ontology with phosphoproteomics and cell imaging to explore the functional impact of CUG-associated spliceopathy on downstream protein metabolism. Changes in phosphorylation, protein isoform expression and intracellular localization in DMSXL astrocytes demonstrate the far-reaching impact of the DM1 repeat expansion on cell metabolism. Our multi-omics approaches provide insight into the mechanisms of CUG RNA toxicity in the CNS with cell type resolution, and support the priority for future research on non-neuronal mechanisms and proteomic changes in DM1 brain disease. Frontiers Media S.A. 2021-05-05 /pmc/articles/PMC8136287/ /pubmed/34025359 http://dx.doi.org/10.3389/fncel.2021.662035 Text en Copyright © 2021 González-Barriga, Lallemant, Dincã, Braz, Polvèche, Magneron, Pionneau, Huguet-Lachon, Claude, Chhuon, Guerrera, Bourgeois, Auboeuf, Gourdon and Gomes-Pereira. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Neuroscience
González-Barriga, Anchel
Lallemant, Louison
Dincã, Diana M.
Braz, Sandra O.
Polvèche, Hélène
Magneron, Paul
Pionneau, Cédric
Huguet-Lachon, Aline
Claude, Jean-Baptiste
Chhuon, Cerina
Guerrera, Ida Chiara
Bourgeois, Cyril F.
Auboeuf, Didier
Gourdon, Geneviève
Gomes-Pereira, Mário
Integrative Cell Type-Specific Multi-Omics Approaches Reveal Impaired Programs of Glial Cell Differentiation in Mouse Culture Models of DM1
title Integrative Cell Type-Specific Multi-Omics Approaches Reveal Impaired Programs of Glial Cell Differentiation in Mouse Culture Models of DM1
title_full Integrative Cell Type-Specific Multi-Omics Approaches Reveal Impaired Programs of Glial Cell Differentiation in Mouse Culture Models of DM1
title_fullStr Integrative Cell Type-Specific Multi-Omics Approaches Reveal Impaired Programs of Glial Cell Differentiation in Mouse Culture Models of DM1
title_full_unstemmed Integrative Cell Type-Specific Multi-Omics Approaches Reveal Impaired Programs of Glial Cell Differentiation in Mouse Culture Models of DM1
title_short Integrative Cell Type-Specific Multi-Omics Approaches Reveal Impaired Programs of Glial Cell Differentiation in Mouse Culture Models of DM1
title_sort integrative cell type-specific multi-omics approaches reveal impaired programs of glial cell differentiation in mouse culture models of dm1
topic Neuroscience
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8136287/
https://www.ncbi.nlm.nih.gov/pubmed/34025359
http://dx.doi.org/10.3389/fncel.2021.662035
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