Reprogramming of dermal fibroblasts from a Duchenne muscular dystrophy patient carrying a deletion of exons 45–50 into an induced pluripotent stem cell line (CCMi005-A)

Rovina, Davide; Castiglioni, Elisa; Mallia, Sara; Rabino, Martina; Farini, Andrea; Belicchi, M.; Giuseppe, Giusy Di; Gervasini, Cristina; Torrente, Yvan; Pompilio, Giulio; Gowran, Aoife

doi:10.1016/j.scr.2022.102889

Cited by 2 publications

(5 citation statements)

References 6 publications

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“…We explored isoforms of dystrophin expressed by hiPSC-derived cardiac fibroblasts (hiPSC-fibs) obtained from control and DMD patients with out of frame deletions of exons (DMD1 and DMD4) and a patient who has a spontaneous point mutation in intron 47 (DMD5) that leads to a frameshift and the formation of a premature stop signal [ 13 – 15 ].…”

Section: Resultsmentioning

confidence: 99%

“…The isolation of the human cells and the subsequent reprogramming into iPSC lines was performed in conformation with the Declaration of Helsinki. iPS cells from 3 DMD patients and 3 healthy donors were obtained from Centro Cardiologico Monzino, Milan Italy, as previously described [ 13 – 15 ]. The three Duchenne patients carried different DMD mutations: two exons deletions and a point mutation.…”

Section: Methodsmentioning

confidence: 99%

“…The three Duchenne patients carried different DMD mutations: two exons deletions and a point mutation. In particular, DMD1 had a deletion of exons 49 and 50 [ 13 ], while DMD4 carried a bigger deletion from exon 45 to exon 50 [ 15 ]. DMD5 carried a point mutation c.6913–1 G > A; this mutation causes the deletion of a nucleotide in the mature transcript with a shift in the reading frame that causes a premature stop codon.…”

Section: Methodsmentioning

confidence: 99%

“…Cell reprogramming into iPSC lines was previously described by Rovina et al [ 15 ]. Briefly, iPSCs were seeded in Matrigel (Corning, 356,230) coated 6 well plates in StemFlex medium (ThermoFisher, A3349401) supplemented with RevitaCell (ThermoFisher, A2644501).…”

Section: Methodsmentioning

confidence: 99%

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IPSC derived cardiac fibroblasts of DMD patients show compromised actin microfilaments, metabolic shift and pro-fibrotic phenotype

Soussi,

Savchenko,

Rovina

et al. 2023

Biol Direct

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Duchenne muscular dystrophy (DMD) is a severe form of muscular dystrophy caused by mutations in the dystrophin gene. We characterized which isoforms of dystrophin were expressed by human induced pluripotent stem cell (hiPSC)-derived cardiac fibroblasts obtained from control and DMD patients. Distinct dystrophin isoforms were observed; however, highest molecular weight isoform was absent in DMD patients carrying exon deletions or mutations in the dystrophin gene. The loss of the full-length dystrophin isoform in hiPSC-derived cardiac fibroblasts from DMD patients resulted in deficient formation of actin microfilaments and a metabolic switch from mitochondrial oxidation to glycolysis. The DMD hiPSC-derived cardiac fibroblasts exhibited a dysregulated mitochondria network and reduced mitochondrial respiration, with enhanced compensatory glycolysis to sustain cellular ATP production. This metabolic remodeling was associated with an exacerbated myofibroblast phenotype and increased fibroblast activation in response to pro fibrotic challenges. As cardiac fibrosis is a critical pathological feature of the DMD heart, the myofibroblast phenotype induced by the absence of dystrophin may contribute to deterioration in cardiac function. Our study highlights the relationship between cytoskeletal dynamics, metabolism of the cell and myofibroblast differentiation and provides a new mechanism by which inactivation of dystrophin in non-cardiomyocyte cells may increase the severity of cardiopathy.

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Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

See 2 more Smart Citations

IPSC derived cardiac fibroblasts of DMD patients show compromised actin microfilaments, metabolic shift and pro-fibrotic phenotype

Soussi,

Savchenko,

Rovina

et al. 2023

Biol Direct

Self Cite

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show abstract

“…PBMCs were reprogrammed to hiPSCs as previously described ( Rovina, Castiglioni et al 2022 ). Briefly, the protocol consisted of electroporation with the reprogramming vectors of the Epi5 Episomal iPSC Reprogramming Kit (ThermoFisher Scientific) at day 0 (1650 V, 10 s, 3 pulse), followed by a gradual transition of the culture medium from StemSpanSFEM II (STEMCELL) to ReproTeSR (STEMCELL) for promoting iPSC colony maturation.…”

Section: Methodsmentioning

confidence: 99%

Generation of four human induced pluripotent stem cell lines from COVID-19 hospitalized patients with increased levels of cardiac Troponin in the acute infection phase developing or not myocarditis

et al. 2023

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Reprogramming of dermal fibroblasts from a Duchenne muscular dystrophy patient carrying a deletion of exons 45–50 into an induced pluripotent stem cell line (CCMi005-A)

Cited by 2 publications

References 6 publications

IPSC derived cardiac fibroblasts of DMD patients show compromised actin microfilaments, metabolic shift and pro-fibrotic phenotype

IPSC derived cardiac fibroblasts of DMD patients show compromised actin microfilaments, metabolic shift and pro-fibrotic phenotype

Generation of four human induced pluripotent stem cell lines from COVID-19 hospitalized patients with increased levels of cardiac Troponin in the acute infection phase developing or not myocarditis

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