Acute conversion of patient-derived Duchenne muscular dystrophy iPSC into myotubes reveals constitutive and inducible over-activation of TGFβ-dependent pro-fibrotic signaling

Caputo, Luca; Granados, Alice; Lenzi, Jessica; Rosa, Alessandro; Ait‐Si‐Ali, Slimane; Puri, Pier Lorenzo; Albini, Sonia

doi:10.1186/s13395-020-00224-7

Cited by 29 publications

(44 citation statements)

References 27 publications

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“…Fibrosis is a major hallmark of DMD pathophysiology, and the regulation of this process has been largely investigated in the past 50,74 . A long‐debated question is the implication of the TGFβ signalling pathway 75,76 . In this study, TGFβ signalling was inhibited up to Day 17 by specific molecules contained in the cell culture media, and TGFβ‐related genes were not up‐regulated at Day 25, suggesting that the observed up‐regulation of fibrosis‐related markers is TGFβ‐independent.…”

Section: Discussionmentioning

confidence: 67%

Myogenesis modelled by human pluripotent stem cells: a multi‐omic study of Duchenne myopathy early onset

Mournetas

Massouridès

Dupont

et al. 2021

J cachexia sarcopenia muscle

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Background Duchenne muscular dystrophy (DMD) causes severe disability of children and death of young men, with an incidence of approximately 1/5000 male births. Symptoms appear in early childhood, with a diagnosis made mostly around 4 years old, a time where the amount of muscle damage is already significant, preventing early therapeutic interventions that could be more efficient at halting disease progression. In the meantime, the precise moment at which disease phenotypes arise—even asymptomatically—is still unknown. Thus, there is a critical need to better define DMD onset as well as its first manifestations, which could help identify early disease biomarkers and novel therapeutic targets. Methods We have used both human tissue‐derived myoblasts and human induced pluripotent stem cells (hiPSCs) from DMD patients to model skeletal myogenesis and compared their differentiation dynamics with that of healthy control cells by a comprehensive multi‐omic analysis at seven time points. Results were strengthened with the analysis of isogenic CRISPR‐edited human embryonic stem cells and through comparisons against published transcriptomic and proteomic datasets from human DMD muscles. The study was completed with DMD knockdown/rescue experiments in hiPSC‐derived skeletal muscle progenitor cells and adenosine triphosphate measurement in hiPSC‐derived myotubes. Results Transcriptome and miRnome comparisons combined with protein analyses demonstrated that hiPSC differentiation (i) leads to embryonic/foetal myotubes that mimic described DMD phenotypes at the differentiation endpoint and (ii) homogeneously and robustly recapitulates key developmental steps—mesoderm, somite, and skeletal muscle. Starting at the somite stage, DMD dysregulations concerned almost 10% of the transcriptome. These include mitochondrial genes whose dysregulations escalate during differentiation. We also describe fibrosis as an intrinsic feature of DMD skeletal muscle cells that begins early during myogenesis. All the omics data are available online for exploration through a graphical interface at https://muscle-dmd.omics.ovh/. Conclusions Our data argue for an early developmental manifestation of DMD whose onset is triggered before the entry into the skeletal muscle compartment, data leading to a necessary reconsideration of dystrophin roles during muscle development. This hiPSC model of skeletal muscle differentiation offers the possibility to explore these functions as well as find earlier DMD biomarkers and therapeutic targets.

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Section: Discussionmentioning

confidence: 67%

Myogenesis modelled by human pluripotent stem cells: a multi‐omic study of Duchenne myopathy early onset

Mournetas

Massouridès

Dupont

et al. 2021

J cachexia sarcopenia muscle

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“…Induced pluripotent stem cell (iPSC) technologies and patient-derived iPSCs provide opportunities for regenerative medicine, drug discovery, and disease modeling from patient-derived stem cells, such as in cystic fibrosis, Huntington’s, Duchenne muscular dystrophy, Alzheimer, amyotrophic lateral sclerosis, age macular degeneration, and retinitis pigmentosa [ 10 , 11 , 12 , 13 , 14 , 15 , 16 ]. Although monolayer cultures are relatively easy to be cultivated, the features that they do not display, the complex tissue organization, limits their bioavailability.…”

Section: Introductionmentioning

confidence: 99%

Expression of Endogenous Angiotensin-Converting Enzyme 2 in Human Induced Pluripotent Stem Cell-Derived Retinal Organoids

Lai

Chou

Chien

et al. 2021

IJMS

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Angiotensin-converting enzyme 2 (ACE2) was identified as the main host cell receptor for the entry of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and its subsequent infection. In some coronavirus disease 2019 (COVID-19) patients, it has been reported that the nervous tissues and the eyes were also affected. However, evidence supporting that the retina is a target tissue for SARS-CoV-2 infection is still lacking. This present study aimed to investigate whether ACE2 expression plays a role in human retinal neurons during SARS-CoV-2 infection. Human induced pluripotent stem cell (hiPSC)-derived retinal organoids and monolayer cultures derived from dissociated retinal organoids were generated. To validate the potential entry of SARS-CoV-2 infection in the retina, we showed that hiPSC-derived retinal organoids and monolayer cultures endogenously express ACE2 and transmembrane serine protease 2 (TMPRSS2) on the mRNA level. Immunofluorescence staining confirmed the protein expression of ACE2 and TMPRSS2 in retinal organoids and monolayer cultures. Furthermore, using the SARS-CoV-2 pseudovirus spike protein with GFP expression system, we found that retinal organoids and monolayer cultures can potentially be infected by the SARS-CoV-2 pseudovirus. Collectively, our findings highlighted the potential of iPSC-derived retinal organoids as the models for ACE2 receptor-based SARS-CoV-2 infection.

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“…This suggests that increased TGFβ release in Gaa KO DBA mice, which are homozygous for the Ltbp4 Δ36 allele, may have an important role in the aetiology of the observed respiratory defects. Differently from PD, the high basal level of TGFβ in the mdx muscle [71] and the superior activation of the TGFβ-SMAD2/3 signalling pathway in MD myotubes [72] may possibly synergize with the increased TGFβ release in the DBA2/J background [42] to significantly worsen MD. Notably, we also found that the early restoration of the missing enzyme in skeletal muscle of Gaa KO DBA mice by systemic administration of AAV-secGAA vectors was able to normalize glycogen content, autophagy/mitophagy block and transcriptome gene expression.…”

Section: Discussionmentioning

confidence: 99%

Gene therapy with secreted acid alpha-glucosidase rescues Pompe disease in a novel mouse model with early-onset spinal cord and respiratory defects

et al. 2020

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Background Pompe disease (PD) is a neuromuscular disorder caused by deficiency of acidalpha-glucosidase (GAA), leading to motor and respiratory dysfunctions. Available Gaa knock-out (KO) mouse models do not accurately mimic PD, particularly its highly impaired respiratory phenotype. Methods Here we developed a new mouse model of PD crossing Gaa KO B6;129 with DBA2/J mice. We subsequently treated Gaa KO DBA2/J mice with adeno-associated virus (AAV) vectors expressing a secretable form of GAA (secGAA). Findings Male Gaa KO DBA2/J mice present most of the key features of the human disease, including early lethality, severe respiratory impairment, cardiac hypertrophy and muscle weakness. Transcriptome analyses of Gaa KO DBA2/J , compared to the parental Gaa KO B6;129 mice, revealed a profoundly impaired gene signature in the spinal cord and a similarly deregulated gene expression in skeletal muscle. Muscle and spinal cord transcriptome changes, biochemical defects, respiratory and muscle function in the Gaa KO DBA2/J model were significantly improved upon gene therapy with AAV vectors expressing secGAA. Interpretation These data show that the genetic background impacts on the severity of respiratory function and neuroglial spinal cord defects in the Gaa KO mouse model of PD. Our findings have implications for PD prognosis and treatment, show novel molecular pathophysiology mechanisms of the disease and provide a unique model to study PD respiratory defects, which majorly affect patients. Funding This work was supported by Genethon, the French Muscular Dystrophy Association (AFM), the European Commission (grant nos. 667751, 617432, and 797144), and Spark Therapeutics.

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Acute conversion of patient-derived Duchenne muscular dystrophy iPSC into myotubes reveals constitutive and inducible over-activation of TGFβ-dependent pro-fibrotic signaling

Cited by 29 publications

References 27 publications

Myogenesis modelled by human pluripotent stem cells: a multi‐omic study of Duchenne myopathy early onset

Myogenesis modelled by human pluripotent stem cells: a multi‐omic study of Duchenne myopathy early onset

Expression of Endogenous Angiotensin-Converting Enzyme 2 in Human Induced Pluripotent Stem Cell-Derived Retinal Organoids

Gene therapy with secreted acid alpha-glucosidase rescues Pompe disease in a novel mouse model with early-onset spinal cord and respiratory defects

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