Long-term maintenance of dystrophin expression and resistance to injury of skeletal muscle in gene-edited mice
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ABSTRACT:
PROVIDER: PRJNA749460 | ENA |
REPOSITORIES: ENA
Dataset's files
| Action | DRS | |||
|---|---|---|---|---|
| SRR15236569.fastq.gz | Fastqsanger.gz | |||
| SRR15236570.fastq.gz | Fastqsanger.gz | |||
| SRR15236571.fastq.gz | Fastqsanger.gz | |||
| SRR15236572.fastq.gz | Fastqsanger.gz | |||
| SRR15236573.fastq.gz | Fastqsanger.gz |
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Project description:Duchenne Muscular Dystrophy (DMD) is a lethal muscle disease caused by mutations in the dystrophin gene. CRISPR/Cas9 genome-editing has been used to correct DMD mutations in animal models at young ages. However, the longevity and durability of CRISPR/Cas9 editing remained to be determined. To address these issues, we subjected DEx44 DMD mice to systemic delivery of AAV9 expressing CRISPR/Cas9 gene editing components to reframe exon 45 of the dystrophin gene, allowing robust dystrophin expression and maintenance of muscle structure and function. We showed that genome correction by CRISPR/Cas9 confers lifelong expression of dystrophin in mice and that corrected skeletal muscle is highly durable and resistant to myofiber necrosis and fibrosis, even in response to chronic injury. In contrast, when muscle fibers were ablated by barium chloride injection, satellite cells were unable to restore dystrophin expression due to restriction of gene editing components. Analysis of on-target and off-target gene editing in aged mice confirmed the stability of gene correction and the lack of significant off-target editing at 18 months of age. These findings demonstrate the long-term durability of CRISPR/Cas9 genome editing as a therapy for maintaining the integrity and function of DMD muscle, even under conditions of stress.
2021-07-27 | GSE180771 | GEO
Project description:Duchenne muscular dystrophy (DMD) is a lethal muscle disease caused by mutations in the dystrophin gene. CRISPR/Cas9 genome editing has been used to correct DMD mutations in animal models at young ages. However, the longevity and durability of CRISPR/Cas9 editing remained to be determined. To address these issues, we subjected ΔEx44 DMD mice to systemic delivery of AAV9-expressing CRISPR/Cas9 gene editing components to reframe exon 45 of the dystrophin gene, allowing robust dystrophin expression and maintenance of muscle structure and function. We found that genome correction by CRISPR/Cas9 confers lifelong expression of dystrophin in mice and that corrected skeletal muscle is highly durable and resistant to myofiber necrosis and fibrosis, even in response to chronic injury. In contrast, when muscle fibers were ablated by barium chloride injection, we observed a loss of gene edited dystrophin expression. Analysis of on- and off-target editing in aged mice confirmed the stability of gene correction and the lack of significant off-target editing at 18 months of age. These findings demonstrate the long-term durability of CRISPR/Cas9 genome editing as a therapy for maintaining the integrity and function of DMD muscle, even under conditions of stress.
| S-EPMC8956962 | biostudies-literature
Project description:We generated and analyzed a conditional Dystrophin flox52/Y: human alpha-skeletal actin muscle knockout (Dmd flox52/Y: HSA mKO) model by targeting exon 52 deletion in the Dystrophin gene to study the consequences of dystrophin loss in skeletal muscle lineages. The generated DMD mouse model ablates dystrophin protein expression after mating with a Cre recombinase transgenic mouse under the control of HSA promoter element that is restricted to the skeletal muscle. The resulting Dmd mKO mice have been assessed using histopathological, phenotypical, functional and biochemical assays based on TRET-NMD standard operating protocols (SOPs) for mdx mouse models. Phenotypic analysis of these conditional Dmd mKO mice revealed a significant decline in locomotor activity and reduced muscle force, motor and muscular function. The histochemical analysis revealed an increase in centralized myonuclei and fibrotic area similar to mdx mice. Immunoassays including western blot and immunohistochemistry confirmed low expression levels of dystrophin in skeletal muscles of Dmd mKO mice. Bulk RNA sequencing analysis revealed that dystrophin loss in myofiber significantly disrupted the expression of cytokines and extracellular matrix genes.
2024-12-25 | GSE284723 | GEO
Project description:Goal was to assess protein linkers between microtubules and dystrophin (specifically dystrophin regions R4-15 and R20-23). Two paired 10-plex TMT experiments were used to compare sixteen unique Dystrophin Glycoprotein Complex (or microtubule) enrichments (four genotypes, each with n=4) plus four pooled samples to allow comparison across the two individual 10-plex experiments for microtubules or DGC. A pair of 10-plex TMT(20 tags total) for the DGC enrichements, and a second pair of TMT (20 tags) for the MT enrichments. Samples are gastroc skeletal muscle from mouse.
2020-09-21 | PXD021088 | Pride
Project description:Molecular profiles of dystophin-deficient patients and normal human skeletal muscles on Affymetrix HG-U95A arrays Keywords = DMD Keywords = Duchenne muscular dystrophy Keywords = dystrophin Keywords = Affymetrix U95A array Keywords = skeletal muscle Keywords = gene expression profiles Keywords: other
2004-01-29 | GSE1004 | GEO
Project description:Genetic mutations in dystrophin manifest in Duchenne muscular dystrophy (DMD), the most prevalent form of a genetically inherited muscle disease. Dystrophin is expressed in skeletal muscle stem cells and fibers, playing a critical role in maintaining skeletal muscle structure, regeneration and function. Here, we report on direct reprogramming of fibroblasts from the Dmdmdx mouse model into induced myogenic progenitor cells (Dmdmdx iMPCs) utilizing transient MyoD overexpression in concert with small molecule treatment. Dmdmdx iMPCs proliferate extensively in vitro and express canonical skeletal muscle stem and progenitor cell markers including Pax7, Sox8 and Myf5. Furthermore, Dmdmdx iMPCs readily give rise to highly contractile and multinucleated myofibers that express a suite of mature skeletal muscle genes however lack dystrophin expression. Utilizing an exon-skipping based approach with CRISPR/Cas9 we report on a genetic correction of the dystrophin mutation in Dmdmdx-iMPCs and subsequent restoration of dystrophin protein expression in vitro. Furthermore, engraftment of genetically-corrected Dmdmdx iMPCs into dystrophic limb muscles of Dmdmdx mice restored dystrophin expression in vivo. Collectively, our findings report on a novel in vitro stem cell-based model for DMD and further establish a new approach to treat this disease via a combination of direct cellular reprogramming, genome engineering and stem cell transplantation methods.
2021-12-15 | GSE164599 | GEO
Project description:Molecular profiles of dystophin-deficient patients and normal human skeletal muscles on Affymetrix HG-U95A arrays Keywords = DMD Keywords = Duchenne muscular dystrophy Keywords = dystrophin Keywords = Affymetrix U95A array Keywords = skeletal muscle Keywords = gene expression profiles Keywords: other
2004-01-29 | E-GEOD-1004 | biostudies-arrayexpress
Project description:Dysfunction of the dystrophin-glycoprotein complex (DGC) is a frequent cause of hereditary forms of muscular dystrophy. Although DGC function in maintaining skeletal muscle integrity has been well characterized, little is known about how the DGC complex is coordinately regulated at the transcriptional level. To test this hypothesis, we engineered HDAC4 stably overexpressing and control myotubes in an in vitro model of muscle differentiation. Here we present evidence that HDAC4, a neural activity-responsive histone deacetylase, is a critical transcriptional regulator of the DGC complex. We show that HDAC4 can repress multiple components of the DGC complex, including dystrophin and sarcoglycan family members in both cultured myotubes. To confirm this finding, the protein levels of core DGC complex members including dystrophin, sarcoglycan complex members, and additional dystrophin-associated proteins were evaluated in differentiated myotubes by western analysis. Keywords: genetic modification and cell type comparison of muscle cells
2008-03-25 | GSE10678 | GEO
Project description:Despite over 3,000 articles published on dystrophin in the last 15 years, the reasons underlying the progression of the human disease, differential muscle involvement, and disparate phenotypes in different species are not understood. The present experiment employed a screen of 12,488 mRNAs in 16-wk-old mouse mdx muscle at a time when the skeletal muscle is avoiding severe dystrophic pathophysiology, despite the absence of a functional dystrophin protein. A number of transcripts whose levels differed between the mdx and human Duchenne muscular dystrophy were noted. A fourfold decrease in myostatin mRNA in the mdx muscle was noted. Differential upregulation of actin-related protein 2/3 (subunit 4), beta-thymosin, calponin, mast cell chymase, and guanidinoacetate methyltransferase mRNA in the more benign mdx was also observed. Transcripts for oxidative and glycolytic enzymes in mdx muscle were not downregulated. These discrepancies could provide candidates for salvage pathways that maintain skeletal muscle integrity in the absence of a functional dystrophin protein in mdx skeletal muscle.
2007-11-06 | E-GEOD-466 | biostudies-arrayexpress
Project description:This study applies targeted Cas9-based gene insertion strategies for the correction of full-length dystrophin in a pre-clinical humanized mouse model of Duchenne muscular dystrophy. Following intramuscular or intravenous delivery, full-length dystrophin is restored in skeletal and cardiac muscle.
2021-04-24 | GSE173224 | GEO