Promising AAV.U7snRNAs vectors targeting DMPK improve DM1 hallmarks in patient-derived cell lines
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ABSTRACT:
ORGANISM(S): Homo sapiens
PROVIDER: GSE233947 | GEO | 2023/06/08
REPOSITORIES: GEO
Similar Datasets
Project description:Myotonic dystrophy type 1 (DM1) is the most common form of muscular dystrophy in adults and affects mainly the skeletal muscle, heart, and brain. DM1 is caused by a CTG repeat expansion in the 3'UTR region of the DMPK gene that sequesters muscleblind-like proteins, blocking their splicing activity and forming nuclear RNA foci. Consequently, many genes have their splicing reversed to a fetal pattern. There is no treatment for DM1, but several approaches have been explored, including antisense oligonucleotides (ASOs) aiming to knock down DMPK expression or bind to the CTGs expansion. ASOs were shown to reduce RNA foci and restore the splicing pattern. However, ASOs have several limitations and although being safe treated DM1 patients did not demonstrate improvement in a human clinical trial. AAV-based gene therapies have the potential to overcome such limitations, providing longer and more stable expression of antisense sequences. In the present study, we designed different antisense sequences targeting exons 5 or 8 of DMPK and the CTG repeat tract aiming to knock down DMPK expression or promote steric hindrance, respectively. The antisense sequences were inserted in U7snRNAs, which were then vectorized in AAV8 particles. Patient-derived myoblasts treated with AAV8. U7snRNAs showed a significant reduction in the number of RNA foci and re-localization of muscle-blind protein. RNA-seq analysis revealed a global splicing correction in different patient-cell lines, without alteration in DMPK expression.
| S-EPMC10309041 | biostudies-literature
Project description:Promising AAV.U7snRNAs vectors targeting DMPK improve DM1 hallmarks in patient-derived cell lines
| PRJNA978624 | ENA
Project description:Recently, the repertoire of human small nucleolar ncRNAs (snoRNAs) and their potential functions have been expanded by the discovery of new snoRNAs and mRNA targets, for which snoRNA-guided modifications may influence their stability, translatability, or splicing. We previously identified snoRNAs that are abundant in healthy human muscle progenitors. Here, we showed that SNORA40 and SNORA70 loss-of-function impairs myogenic differentiation. Strikingly, their gain-of-function can rescue the impaired differentiation muscle progenitor cells in Myotonic Dystrophy type 1 (DM1). We identified the CCND3 mRNA, partly located in the nucleolus, as a target of SNORA40 and SNORA70, which are needed for its pseudouridylated status. Expression of the CCND3 protein is required for muscle progenitors to exit the cell cycle when they are induced to differentiation. Here, we revealed that this switch requires SNORA40/70. Finally, we showed that rescuing reduced levels found in DM1 cells restores the expression of CCND3 through pseudouridylation of its mRNA, and ultimately resumes the cell fusion capacity of DM1 muscle progenitors.
2025-03-31 | GSE227349 | GEO
Project description:Recently, the repertoire of human small nucleolar noncoding RNAs (snoRNAs) and their potential functions has expanded with the discovery of new snoRNAs and messenger RNA (mRNA) targets, for which snoRNA-guided modifications may influence their stability, translatability, and splicing. We previously identified snoRNAs that are abundant in healthy human muscle progenitor cells. In this study, we demonstrated that SNORA40 and SNORA70 loss-of-function impairs myogenic differentiation. Interestingly, gain-of-function can rescue impaired differentiation muscle progenitor cells in myotonic dystrophy type 1 (DM1). We identified cyclin D3 (CCND3) mRNA, which is partially located in the nucleolus, as a target for SNORA40 and SNORA70, which are required for its pseudouridylated status. Expression of the CCND3 protein is required for muscle progenitors to exit the cell-cycle when they are induced to differentiate. We revealed that this switch requires SNORA40/70. Finally, we observed that DM1 cells show reduced levels of SNORA40/70 and undetectable CCND3 protein. However, restoring normal levels of SNORA40/70 partially restored CCND3 protein expression, coinciding with improved cell fusion capacity in DM1 muscle progenitors. Collectively, these data suggest that this effect may stem from SNORA40/70-dependent pseudouridylation of CCND3 mRNA, emphasizing snoRNAs as key players in normal and pathological muscle differentiation.
2025-03-31 | GSE269301 | GEO
Project description:The genetic cause of Myotonic Dystrophy Type 1 (DM1) is the expansion of unstable CTG-repeats in the 3´ untranslated region of the DMPK gene. Therapeutic gene editing for treatment of monogenic diseases could in principle remove definitively the disease-causing genetic defect. Here we describe the application of the CRISPR/Cas9 strategy to remove the CTG-expansion in the DMPK gene in myogenic cells derived from a DM1 patient that is heterozygous for the expansion. The efficacy of on-target genomic events was assessed by performing long-read DNA sequencing (Oxford Nanopore Technologies) after applying Cas9-mediated target enrichment of the Regions Of Interest (ROI), enabling the simultaneous analysis (a) of cuts by both sgRNAs resulting in the effective excision of the CTG expansion, (b) of cuts induced only by a single sgRNA, (c) of the potential occurrence of aberrant events such as inversions, and (d) the existence of non-edited sequences, within the same sample.
2025-02-26 | GSE279054 | GEO
Project description:In this Study, we used RNA-targeting Cas9 (RCas9) to reverse characteristic Myotonic Dystrophy (DM1) cellular phenotypes such as elimination of RNA foci, MBNL relocalization, and reversal of transcriptome-wide splicing in a mouse model of myotonic Dystrophy (DM1). Furthermore we show that gene expression is not altered with RCas9 treatment in WT mice with or without treatment with immunosuppression
2020-07-01 | GSE152033 | GEO
Project description:Misregulated alternative splicing appears to be a major factor in the pathogenesis of myotonic dystrophy. The present study was done to further explore alternative splicing in this condition by doing exon-level analysis of mRNA from skeletal muscle of 8 subjects with type 1 myotonic dystrophy, 7 subjects with type 2 myotonic dystrophy, 8 disease controls (subjects with facioscapulohumeral muscular dystrophy), and 8 healthy controls . The ratios of signals from the various exons of a gene provided an index of altered exon inclusion/exclusion that was independent of the overall expression of that gene. There were numerous transcripts for which there was evidence of abnormal alternative splicing in subjects with myotonic dystrophy. For many of these transcripts, the abnormal splicing was confirmed by an independent RT-PCR approach. 31 subjects, one sample per subject, four groups: healthy subjects (n = 8), facioscapulohumeral dystrophy (n = 8), type 1 myotonic dystrophy (n = 8), type 2 myotonic dystrophy (n = 7)
2013-06-15 | E-GEOD-47968 | biostudies-arrayexpress
Project description:Comparison of myotonic dystrophy cataract samples with control non-myotonic dystrophy cataract samples
2011-10-31 | E-MEXP-3365 | biostudies-arrayexpress
Project description:Myotonic dystrophy type 1 (DM1) is a multisystem genetic disorder involving the muscle, heart, and central nervous system (CNS). The pathogenesis of CNS symptoms prevalent in patients with DM1 remains unelucidated. To elucidate the CNS pathogenesis in DM1, we investigated cell type-specific abnormalities in cortical neurons, white matter glial cells, and spinal motor neurons of patients with DM1 via laser-capture microdissection(LCM).
2022-07-07 | GSE198321 | GEO