Main description:

This detailed book presents a comprehensive collection of state-of-the-art protocols on muscular dystrophy therapeutics, covering therapeutics using antisense oligonucleotides, gene replacement, genome editing, small molecules, stem cells, and antibodies. Written by leaders in the field, the volume explores techniques that are currently in use and are starting an exciting therapeutic revolution in muscular dystrophy. As a part of the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step and readily reproducible laboratory protocols, as well as tips on troubleshooting and avoiding known pitfalls.
Authoritative and practical, Muscular Dystrophy Therapeutics: Methods and Protocols serves as an ideal resource to inspire readers and provide tips, strategies, and advice to develop new therapeutic technologies for this group of diseases.

Contents:

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

15. Molecular and Biochemical Assessment of Gene Therapy in the Canine Model of Duchenne Muscular Dystrophy

Chady H. Hakim, Dennis Perez-Lopez, Matt Burke, James Teixeira, and Dongsheng Duan

16. Histological Assessment of Gene Therapy in the Canine DMD Model

Chady H. Hakim, Matt Burke, James Teixeira, and Dongsheng Duan

17. MRI Evaluation of Gene Therapy in the Canine Model of Duchenne Muscular Dystrophy

Amy R. Zalcman, Chady H. Hakim, Jimmy Lattimer, James R. Holland, John R. Dodam, and Dongsheng Duan

18. Assessment of the Gene Therapy Immune Response in the Canine Muscular Dystrophy Model

Chady H. Hakim, Sandeep R.P. Kumar, Dennis Perez-Lopez, James Teixeira, Roland W. Herzog, and Dongsheng Duan

19. Use of Mesenchymal Stem Cells to Enhance the Efficacy of Gene Therapy

Hiromi Hayashita-Kinoh and Takashi Okada

20. Exon-Skipping for a Pathogenic COL6A1 Variant in Ullrich CMD

Sara Aguti, Fady Guirguis, Carsten Boennemann, Francesco Muntoni, Veronique Bolduc, and Haiyan Zhou

Part V: Genome Editing/CRISPR

21. CRISPR-Cas9 Correction of Duchenne Muscular Dystrophy in Mice by a Self-Complementary AAV Delivery System

Yu Zhang, Rhonda Bassel-Duby, and Eric N. Olson

22. Preparation of NanoMEDIC Extracellular Vesicles to Deliver CRISPR-Cas9 Ribonucleoproteins for Genomic Exon Skipping

Kei Watanabe, Peter Gee, and Akitsu Hotta

23. Restoration of Dystrophin Expression in Mdx-Derived Muscle Progenitor Cells Using CRISPR/Cas9 System and Homology-Directed Repair Technology

Yue Jin, Yan Shen, Il-man Kim, Neal L. Weintraub, Mark Hamrick, and Yaoliang Tang

Part VI: Small Molecules

24. Effects of Glucocorticoids in Murine Models of Duchenne and Limb-Girdle Muscular Dystrophy

Michelle Wintzinger, Karen Miz, Allen York, Alexis R. Demonbreun, Jeffery D. Molkentin, Elizabeth M. McNally, and Mattia Quattrocelli

25. High-Throughput Screening to Identify Modulators of Sarcospan

Cynthia Shu, Ekaterina Mokhonova, and Rachelle H. Crosbie

26. Identifying FDA-Approved Drugs that Upregulate Utrophin A as a Therapeutic Strategy for Duchenne Muscular Dystrophy

Christine Peladeau and Bernard J. Jasmin

Part VII: Cell Models and Stem Cells

27. Monitoring Membrane Injury-Triggered Endocytosis at Single Cell and Single Vesicle Resolution

Daniel C. Bittel and Jyoti K. Jaiswal

28. Evaluation of hiPSC-Derived Muscle Progenitor Cell Transplantation in a Mouse Duchenne Muscular Dystrophy Model

Minas Nalbandian, Mingming Zhao, and Hidetoshi Sakurai

29. Quantification of Muscle Satellite Stem Cell Divisions by High Content Analysis

William Chen, Theodore J. Perkins, and Michael A. Rudnicki

Part VIII: Antibody

30. Systemic Delivery of a Monoclonal Antibody to Immunologically Block Myostatin in the A17 Mouse Model of OPMD

Alberto Malerba, Pradeep Harish, and Linda Popplewell