Muscular Dystrophy Therapeutics: Methods and Protocols(2587 Methods in Molecular Biology)

Part I: Basics and Introduction 1. Current Strategies of Muscular Dystrophy Therapeutics: An Overview Kenji Rowel Q. Lim and Toshifumi Yokota 2. The Story of Viltolarsen: From Preclinical Studies to FDA Approval Rohini Roy Roshmi and Toshifumi Yokota Part II: Sample Preparation and Assessment 3. Rapid Freezing of Skeletal and Cardiac Muscles Using Isopentane Cooled with Liquid Nitrogen and Tragacanth Gum for Histological, Genetic, and Protein Expression Studies Saeed Anwar and Toshifumi Yokota 4. Cardiac and Skeletal Muscle Pathology in the D2/mdx Mouse Model and Caveats Associated with the Quantification of Utrophin Tahnee L. Kennedy and Hannah F. Dugdale 5. Physiological Assessment of Muscle, Heart, and Whole Body Function in the Canine Model of Duchenne Muscular Dystrophy Chady H. Hakim, James Teixeira, Stacy Leach, and Dongsheng Duan Part III: Antisense Oligonucleotides 6. Restoring Dystrophin Expression by Skipping Exon 6 and 8 in Neonatal Dystrophic Dogs Md Nur Ahad Shah and Toshifumi Yokota 7. Restoring Dystrophin Expression with Exon 44 and 53 Skipping in the DMD Gene in Immortalized Myotubes Yusuke Echigoya and Toshifumi Yokota 8. Restoring Dystrophin Expression with Duchenne Muscular Dystrophy Exon 45 Skipping in Induced-Pluripotent Stem Cell-Derived Cardiomyocytes Mitsuto Sato, Naoko Shiba, Daigo Miyazaki, Yuji Shiba, and Akinori Nakamura 9. Quantitative Evaluation of Exon Skipping in Urine-Derived Cells for Duchenne Muscular Dystrophy Katsuhiko Kunitake, Chaitra Sathyaprakash, Norio Motohashi, and Yoshitsugu Aoki 10. Use of Glycine to Augment Exon Skipping and Cell Therapies for Duchenne Muscular Dystrophy Gang Han, Caorui Lin, and HaiFang Yin 11. Morpholino-Mediated Exons 28-29 Skipping in Dysferlin Saeed Anwar and Toshifumi Yokota 12. Knocking Down DUX4 in Immortalized Facioscapulohumeral Muscular Dystrophy Patient-Derived Muscle Cells Kenji Rowel Q. Lim and Toshifumi Yokota 13. Peptide-Conjugated PMOs for the Treatment of Myotonic Dystrophy Jessica Stoodley, David Seone Miraz, Yahya Jad, Mathieu Fischer, Matthew J.A. Wood, and Miguel A. Varela 14. Developing Therapeutic Splice-Correcting Antisense Oligomers for Adult-Onset Pompe Disease with c.-32-13T>G Mutation Kristin A. Ham, Russell D. Johnsen, Michel Tchan, Steve D. Wilton, and May T. Aung-Htut Part IV: Gene Replacement Therapies

Table of Contents:
Current Strategies of Muscular Dystrophy Therapeutics: An Overview.- The Story of Viltolarsen: From Preclinical Studies to FDA Approval.- Rapid Freezing of Skeletal and Cardiac Muscles Using Isopentane Cooled with Liquid Nitrogen and Tragacanth Gum for Histological, Genetic, and Protein Expression Studies.- Cardiac and Skeletal Muscle Pathology in the D2/mdx Mouse Model and Caveats Associated with the Quantification of Utrophin.- Physiological Assessment of Muscle, Heart, and Whole Body Function in the Canine Model of Duchenne Muscular Dystrophy.- Restoring Dystrophin Expression by Skipping Exon 6 and 8 in Neonatal Dystrophic Dogs.- Restoring Dystrophin Expression with Exon 44 and 53 Skipping in the DMD Gene in Immortalized Myotubes.- Restoring Dystrophin Expression with Duchenne Muscular Dystrophy Exon 45 Skipping in Induced-Pluripotent Stem Cell-Derived Cardiomyocytes.- Quantitative Evaluation of Exon Skipping in Urine-Derived Cells for Duchenne Muscular Dystrophy.- Use of Glycine to Augment Exon Skipping and Cell Therapies for Duchenne Muscular Dystrophy.- Morpholino-Mediated Exons 28–29 Skipping in Dysferlin.- Knocking Down DUX4 in Immortalized Facioscapulohumeral Muscular Dystrophy Patient-Derived Muscle Cells.- Peptide-Conjugated PMOs for the Treatment of Myotonic Dystrophy.- Developing Therapeutic Splice-Correcting Antisense Oligomers for Adult-Onset Pompe Disease with c.-32-13T>G Mutation.- Molecular and Biochemical Assessment of Gene Therapy in the Canine Model of Duchenne Muscular Dystrophy.- Histological Assessment of Gene Therapy in the Canine DMD Model.- MRI Evaluation of Gene Therapy in the Canine Model of Duchenne Muscular Dystrophy.- Assessment of the Gene Therapy Immune Response in the Canine Muscular Dystrophy Model.- Use of Mesenchymal Stem Cells to Enhance the Efficacy of Gene Therapy.- Exon-Skipping for a Pathogenic COL6A1 Variant in Ullrich CMD.- CRISPR-Cas9 Correction of Duchenne Muscular Dystrophy in Mice by a Self-Complementary AAV Delivery System.- Preparation of NanoMEDIC Extracellular Vesicles to Deliver CRISPR-Cas9 Ribonucleoproteins for Genomic Exon Skipping.- Restoration of Dystrophin Expression in Mdx-Derived Muscle Progenitor Cells Using CRISPR/Cas9 System and Homology-Directed Repair Technology.- Effects of Glucocorticoids in Murine Models of Duchenne and Limb-Girdle Muscular Dystrophy.- High-Throughput Screening to Identify Modulators of Sarcospan.- Identifying FDA-Approved Drugs that Upregulate Utrophin A as a Therapeutic Strategy for Duchenne Muscular Dystrophy.- Monitoring Membrane Injury-Triggered Endocytosis at Single Cell and Single Vesicle Resolution.- Evaluation of hiPSC-Derived Muscle Progenitor Cell Transplantation in a Mouse Duchenne Muscular Dystrophy Model.- Quantification of Muscle Satellite Stem Cell Divisions by High Content Analysis.- Systemic Delivery of a Monoclonal Antibody to Immunologically Block Myostatin in the A17 Mouse Model of OPMD.