CRISPR-Cas9 editing of a TNPO3 mutation in a muscle cell model of limb-girdle muscular dystrophy type D2.

Fecha de publicación: 14/03/2023 Fecha Ahead of Print: 11/01/2023

Autores de IIS La Fe

Javier Poyatos García

Autor
Marta Selva Gimenez

Autor
Ariadna Bargiela Schonbrunn

Autor
Rafael Vázquez Manrique

Autor
Juan Jesús Vílchez Padilla

Autor

Participantes ajenos a IIS La Fe

Blazquez-Bernal, Agueda
Espinosa-Espinosa, Jorge
Bigot, Anne
Artero, Ruben

Grupos

Biomedicina Molecular, Celular y Genómica

Leader

José María Millán Salvador
Patología Neuromuscular y Ataxias

Investigador Principal

Teresa Sevilla Mantecón
Unidad mixta de investigación en enfermedades raras (CIPF)

Leader

José María Millán Salvador

Abstract

A single-nucleotide deletion in the stop codon of the nuclear import receptor transportin-3 (TNPO3), also involved in human immunodeficiency virus type 1 (HIV-1) infection, causes the ultrarare autosomal dominant disease limb-girdle muscular dystrophy D2 (LGMDD2) by extending the wild-type protein. Here, we generated a patient-derived in vitro model of LGMDD2 as an immortalized myoblast cell line carrying the TNP O 3 mutation. The cell model reproduced critical molecular alterations seen in patients, such as TNP O 3 overexpression, defects in terminal muscle markers, and autophagy overactivation. Correction of the TNP O 3 mutation via CRISPR-Cas9 editing caused a significant reversion of the pathological phenotypes in edited cells, including a complete absence of the mutant TNPO3 protein, as detected with a polyclonal antibody specific against the abnormal 15-aa peptide. Transcriptomic analyses found that 15% of the transcriptome was differentially expressed in model myotubes. CRISPR-Cas9-corrected cells showed that 44% of the alterations were rescued toward normal levels. MicroRNAs (miRNAs) analyses showed that around 50% of miRNAs with impaired expression because of the disease were recovered on the mutation edition. In summary, this work provides proof of concept of the potential of CRISPR-Cas9-mediated gene editing of TNP O 3 as a therapeutic approach and describes critical reagents in LGMDD2 research.

Datos de la publicación

ISSN/ISSNe:: 2162-2531, 2162-2531
Tipo:: Article
Páginas:: 324-338
DOI:: 10.1016/j.omtn.2023.01.004
Factor de Impacto:: 1,852 SCImago ℠
Cuartil:: Q1 SCImago ℠

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Métricas

Filiaciones

Poyatos-Garcia, Javier:: Centre for Biomedical Network Research on Rare Diseases (CIBERER), U763, CB06/05/0091, 46026 Valencia, Spain

Instituto de Investigación. Neuromuscular and Ataxias Research Group, Health Research Institute Hospital La Fe (IIS La Fe), 46026 Valencia, Spain
Blazquez-Bernal, Agueda:: Translational Genomics Group, University Institute for Biotechnology and Biomedicine (BIOTECMED), University of Valencia, Burjasot, 46100 Valencia, Spain

INCLIVA Biomedical Research Institute, 46010 Valencia, Spain
Selva-Gimenez, Marta:: Centre for Biomedical Network Research on Rare Diseases (CIBERER), U763, CB06/05/0091, 46026 Valencia, Spain

Instituto de Investigación. Neuromuscular and Ataxias Research Group, Health Research Institute Hospital La Fe (IIS La Fe), 46026 Valencia, Spain
Bargiela, Ariadna:: Instituto de Investigación. Neuromuscular and Ataxias Research Group, Health Research Institute Hospital La Fe (IIS La Fe), 46026 Valencia, Spain
Espinosa-Espinosa, Jorge:: Translational Genomics Group, University Institute for Biotechnology and Biomedicine (BIOTECMED), University of Valencia, Burjasot, 46100 Valencia, Spain

INCLIVA Biomedical Research Institute, 46010 Valencia, Spain
Vazquez-Manrique, Rafael P:: Centre for Biomedical Network Research on Rare Diseases (CIBERER), U763, CB06/05/0091, 46026 Valencia, Spain

Instituto de Investigación. Grupo de Investigación en Biomedicina Molecular, Celular y Genómica, Instituto de Investigación Sanitaria La Fe (IIS La Fe), 46026 Valencia, Spain

Instituto de Investigación. Joint Unit for Rare Diseases IIS La Fe-CIPF, 46012 Valencia, Spain
Bigot, Anne:: Sorbonne Université, INSERM, Institut de Myologie, Centre de Recherche en Myologie, 75013 Paris, France
Artero, Ruben:: Translational Genomics Group, University Institute for Biotechnology and Biomedicine (BIOTECMED), University of Valencia, Burjasot, 46100 Valencia, Spain

INCLIVA Biomedical Research Institute, 46010 Valencia, Spain
Vilchez, Juan Jesus:: Centre for Biomedical Network Research on Rare Diseases (CIBERER), U763, CB06/05/0091, 46026 Valencia, Spain

Instituto de Investigación. Neuromuscular and Ataxias Research Group, Health Research Institute Hospital La Fe (IIS La Fe), 46026 Valencia, Spain