Genome Editing-Mediated Utrophin Upregulation in Duchenne Muscular Dystrophy Stem Cells
| العنوان: | Genome Editing-Mediated Utrophin Upregulation in Duchenne Muscular Dystrophy Stem Cells |
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| المؤلفون: | Manoj K. Mishra, Emanuele Loro, Kasturi Sengupta, Melissa J. Spencer, Tejvir S. Khurana, April D. Pyle |
| المصدر: | Molecular Therapy. Nucleic Acids Molecular Therapy: Nucleic Acids, Vol 22, Iss, Pp 500-509 (2020) |
| سنة النشر: | 2020 |
| مصطلحات موضوعية: | Untranslated region, musculoskeletal diseases, Duchenne muscular dystrophy, congenital, hereditary, and neonatal diseases and abnormalities, utrophin, MiRNA binding, Downregulation and upregulation, Genome editing, DMD-hiPSC, stem cells, Drug Discovery, Utrophin, microRNA, medicine, IMTR, biology, UTRNΔIMTR, lcsh:RM1-950, medicine.disease, musculoskeletal system, gene therapy, Cell biology, lcsh:Therapeutics. Pharmacology, biology.protein, Molecular Medicine, Original Article, CRISPR-Cas9, Dystrophin |
| الوصف: | Utrophin upregulation is considered a promising therapeutic strategy for Duchenne muscular dystrophy (DMD). A number of microRNAs (miRNAs) post-transcriptionally regulate utrophin expression by binding their cognate sites in the 3′ UTR. Previously we have shown that miRNA: UTRN repression can be alleviated using miRNA let-7c site blocking oligonucleotides (SBOs) to achieve utrophin upregulation and functional improvement in mdx mice. Here, we used CRISPR/Cas9-mediated genome editing to delete five miRNA binding sites (miR-150, miR-296-5p, miR-133b, let-7c, miR-196b) clustered in a 500 bp inhibitory miRNA target region (IMTR) within the UTRN 3′ UTR, for achieving higher expression of endogenous utrophin. Deleting the UTRN IMTR in DMD patient-derived human induced pluripotent stem cells (DMD-hiPSCs) resulted in ca. 2-fold higher levels of utrophin protein. Differentiation of the UTRN edited DMD-hiPSCs (UTRNΔIMTR) by MyoD overexpression resulted in increased sarcolemmal α-sarcoglycan staining consistent with improved dystrophin glycoprotein complex (DGC) restoration. These results demonstrate that CRISPR/Cas9-based UTRN genome editing offers a novel utrophin upregulation therapeutic strategy applicable to all DMD patients, irrespective of the dystrophin mutation status. Graphical Abstract Sengupta et al. report a genome editing strategy for upregulating utrophin in Duchenne muscular dystrophy patient iPSCs. Edited iPSC-derived myotubes demonstrate increased expression of utrophin and components of the dystroglycan/sarcoglycan complex. This approach represents a promising therapeutic strategy applicable for all DMD patients, irrespective of their mutation status. |
| تدمد: | 2162-2531 |
| URL الوصول: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::a5060a4d573f9dd9565b3189d6345a64 https://pubmed.ncbi.nlm.nih.gov/33230452 |
| Rights: | OPEN |
| رقم الانضمام: | edsair.doi.dedup.....a5060a4d573f9dd9565b3189d6345a64 |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 21622531 |
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