{
  "abstract": "Background: Duchenne muscular dystrophy (DMD) is a X-linked disease affecting skeletal and cardiac muscle and is caused by mutations in the dystrophin gene (DMD). Patient-derived induced pluripotent stem cells (iPSCs) serve as reliable in vitro disease models. Their genetic correction by CRISPR/Cas9 allows the generation of isogenic controls and holds promises for gene therapy. However, restoring full-length dystrophin, especially when deletions involve multiple exons, constitutes a technological challenge. This study aimed to fully repair the dystrophin gene from a DMD iPSC line carrying the deletion of exons 49–50 and to characterize the rescue of the cardiac phenotype.\n\nMethods: We developed an innovative CRISPR/Cas9-based approach involving the insertion of coding sequences of the deleted region, at the 3’ of exon 48, thereby generating a single continuous coding sequence encompassing exons 48-49-50. Subsequently, iPSCs were differentiated into cardiomyocytes and cardiac fibroblasts. Cardiac phenotypes were analysed by western blot, immunofluorescence, ELISA, FACS, Ionoptix, 3D engineered heart tissue (EHT) and single-nuclei RNA-seq.\n\nResults: The correction of a two-exons DMD gene deletion in Duchenne iPSCs, using CRISPR/Cas9, enabled the re-expression of a stable and functional full-length dystrophin in cardiomyocytes resulting in the rescue of cardiac pathological phenotypes. Edited cardiomyocytes showed improved morphology, reduced release of the cardiomyocytes damage marker troponin I, and decreased ROS production. Moreover, dystrophin restoration enhanced contractility and ameliorated the Ca2+ kinetics. Notably, edited iPSC derived fibroblasts showed reduced pro-fibrotic stimuli response. In parallel, we also observed enhanced functioning of a 3D engineered heart tissue and profound change in the transcriptomic profile in both cardiomyocytes and fibroblasts after the re-expression of full-length dystrophin.\n\nConclusions: We developed an innovative approach that enabled the re-expression of full-length dystrophin in a DMD iPSC line with consequent complete rescue of in vitro DMD cardiac phenotypes. On the long term, these results could lay a foundation for future applications of cell therapy or in vivo CRISPR/Cas9-based intervention to treat DMD.\n\nSupplementary Information: The online version contains supplementary material available at https://doi.org/10.1186/s13073-026-01738-3.",
  "abstract_shown": true,
  "affiliations": [
    "Unit of Inherited Cardiomyopathies, Centro Cardiologico Monzino IRCCS, Milan, 20138 Italy",
    "iPSC Facility for Cardiac Modelling and Function, Centro Cardiologico Monzino IRCCS, Milan, 20138 Italy",
    "Department of Pharmacy, School of Medicine and Surgery, University of Naples Federico II, Naples, 80131 Italy",
    "Department of Biology, University of Florence, Florence, 50134 Italy",
    "Department of NeuroFarBa, University of Florence, Florence, 50139 Italy",
    "Department of Experimental and Clinical Medicine, University of Florence, Florence, 50139 Italy",
    "Bioinformatics and Artificial Intelligence Facility, Centro Cardiologico Monzino IRCCS, Milan, 20138 Italy",
    "Department of Electronics, Information and Biomedical Engineering, Politecnico di Milano, Milan, 20133 Italy",
    "Department of Thoracic and Cardiovascular Diseases, Azienda Ospedaliero-Universitaria Maggiore della Carità, Novara, 28100 Italy",
    "Department of Translational Medicine, University of Eastern Piedmont, Novara, 28100 Italy",
    "Paediatric Neurology and Centro Clinico NeMO, Policlinico Gemelli, Catholic University, Fondazione Policlinico Universitario Agostino Gemelli IRCSS, Rome, 00168 Italy",
    "Department of Biomedical, Surgical and Dental Sciences, Università degli Studi di Milano, Milan, 20122 Italy"
  ],
  "article_type": "research article",
  "arxiv": null,
  "author_total": 15,
  "authors": [
    {
      "affs": [
        0,
        1
      ],
      "name": "Davide Rovina"
    },
    {
      "affs": [
        0,
        2
      ],
      "name": "Sofia Milasi"
    },
    {
      "affs": [
        0
      ],
      "name": "Giusy Di Giuseppe"
    },
    {
      "affs": [
        3
      ],
      "name": "Josè Manuel Pioner"
    },
    {
      "affs": [
        3
      ],
      "name": "Marianna Langione"
    },
    {
      "affs": [
        4
      ],
      "name": "Lucrezia Giammarino"
    },
    {
      "affs": [
        5
      ],
      "name": "Cecilia Ferrantini"
    },
    {
      "affs": [
        6,
        7
      ],
      "name": "Mattia Chiesa"
    },
    {
      "affs": [
        8
      ],
      "name": "Francesco Canonico"
    },
    {
      "affs": [
        9
      ],
      "name": "Domenico D’Amario"
    },
    {
      "affs": [
        10
      ],
      "name": "Eugenio Maria Mercuri"
    },
    {
      "affs": [
        0,
        11
      ],
      "name": "Giulio Pompilio"
    },
    {
      "affs": [
        0
      ],
      "name": "Elena Sommariva"
    },
    {
      "affs": [
        0
      ],
      "name": "Aoife Gowran"
    },
    {
      "affs": [
        1
      ],
      "name": "Angela Serena Maione"
    }
  ],
  "bibtex": "https://research.draconitic.com/paper/79z15nzsjjxr0.bib",
  "cited_by": 0,
  "doi": "10.1186/s13073-026-01738-3",
  "fulltext_url": "https://pmc.ncbi.nlm.nih.gov/articles/PMC13715161/",
  "id": "79z15nzsjjxr0",
  "issns": [
    "1756-994X"
  ],
  "landing_url": "https://doi.org/10.1186/s13073-026-01738-3",
  "language": "en",
  "licence": "CC-BY-NC-ND-4.0",
  "licence_label": "CC BY-NC-ND 4.0",
  "licence_source": "article",
  "licence_url": "https://creativecommons.org/licenses/by-nc-nd/4.0/",
  "metadata_licence": null,
  "native_subjects": [
    [
      "heading",
      "Research"
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      "openalex",
      "1312|CRISPR and Genetic Engineering"
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  "openalex": "W7172014218",
  "pdf_url": null,
  "pmcid": "PMC13715161",
  "pmid": "42947239",
  "published": "2026-07-31",
  "publisher": "BMC",
  "ris": "https://research.draconitic.com/paper/79z15nzsjjxr0.ris",
  "sources": [
    {
      "first_seen": "2026-11-01T12:22:31Z",
      "native_id": "PMC13715161",
      "source": "pmc",
      "url": "https://doi.org/10.1186/s13073-026-01738-3"
    }
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  "subjects": [
    {
      "field": "biochemistry",
      "origin": "openalex:1312|CRISPR and Genetic Engineering",
      "subfield": "molecular-biology"
    }
  ],
  "title": "Phenotype rescue through the restoration of full-length dystrophin using CRISPR/Cas9 genome editing in Duchenne muscular dystrophy patient-derived iPSCs carrying the deletion of two exons",
  "title_text": "Phenotype rescue through the restoration of full-length dystrophin using CRISPR/Cas9 genome editing in Duchenne muscular dystrophy patient-derived iPSCs carrying the deletion of two exons",
  "type": "peer-reviewed",
  "updated_at": "2026-11-01T12:22:31Z",
  "url": "https://research.draconitic.com/paper/79z15nzsjjxr0",
  "venue": "Genome Medicine",
  "versions": []
}