scholarly journals Toward the correction of muscular dystrophy by gene editing

2021 ◽  
Vol 118 (22) ◽  
pp. e2004840117
Author(s):  
Eric N. Olson
Keyword(s):  
Muscular Dystrophy ◽  
Gene Editing ◽  
Genetic Disorder ◽  
Premature Death ◽  
Lessons Learned ◽  
Large Mammals ◽  
Gene Encoding ◽  
Monogenic Disorders ◽  
Dystrophin Expression ◽  
And Function

Recent advances in gene editing technologies are enabling the potential correction of devastating monogenic disorders through elimination of underlying genetic mutations. Duchenne muscular dystrophy (DMD) is an especially severe genetic disorder caused by mutations in the gene encoding dystrophin, a membrane-associated protein required for maintenance of muscle structure and function. Patients with DMD succumb to loss of mobility early in life, culminating in premature death from cardiac and respiratory failure. The disease has thus far defied all curative strategies. CRISPR gene editing has provided new opportunities to ameliorate the disease by eliminating DMD mutations and thereby restore dystrophin expression throughout skeletal and cardiac muscle. Proof-of-concept studies in rodents, large mammals, and human cells have validated the potential of this approach, but numerous challenges remain to be addressed, including optimization of gene editing, delivery of gene editing components throughout the musculature, and mitigation of possible immune responses. This paper provides an overview of recent work from our laboratory and others toward the genetic correction of DMD and considers the opportunities and challenges in the path to clinical translation. Lessons learned from these studies will undoubtedly enable further applications of gene editing to numerous other diseases of muscle and other tissues.

scholarly journals CRISPR Correction of Duchenne Muscular Dystrophy

2019 ◽  
Vol 70 (1) ◽  
pp. 239-255 ◽  
Author(s):  
Yi-Li Min ◽  
Rhonda Bassel-Duby ◽  
Eric N. Olson
Keyword(s):  
Duchenne Muscular Dystrophy ◽  
Muscular Dystrophy ◽  
Recent Progress ◽  
Scaffolding Protein ◽  
Genetic Mutations ◽  
Striated Muscles ◽  
Gene Encoding ◽  
Monogenic Disorders ◽  
The Third ◽  
Dystrophin Expression

The ability to efficiently modify the genome using CRISPR technology has rapidly revolutionized biology and genetics and will soon transform medicine. Duchenne muscular dystrophy (DMD) represents one of the first monogenic disorders that has been investigated with respect to CRISPR-mediated correction of causal genetic mutations. DMD results from mutations in the gene encoding dystrophin, a scaffolding protein that maintains the integrity of striated muscles. Thousands of different dystrophin mutations have been identified in DMD patients, who suffer from a loss of ambulation followed by respiratory insufficiency, heart failure, and death by the third decade of life. Using CRISPR to bypass DMD mutations, dystrophin expression has been efficiently restored in human cells and mouse models of DMD. Here, we review recent progress toward the development of possible CRISPR therapies for DMD and highlight opportunities and potential obstacles in attaining this goal.

scholarly journals Gene editing restores dystrophin expression in a canine model of Duchenne muscular dystrophy

Science ◽  
2018 ◽  
Vol 362 (6410) ◽  
pp. 86-91 ◽  
Author(s):  
Leonela Amoasii ◽  
John C. W. Hildyard ◽  
Hui Li ◽  
Efrain Sanchez-Ortiz ◽  
Alex Mireault ◽  
...  
Keyword(s):  
Duchenne Muscular Dystrophy ◽  
Muscular Dystrophy ◽  
Gene Editing ◽  
Canine Model ◽  
Large Animal ◽  
Systemic Delivery ◽  
Gene Encoding ◽  
Animal Data ◽  
And Function ◽  
Dystrophin Protein

Mutations in the gene encoding dystrophin, a protein that maintains muscle integrity and function, cause Duchenne muscular dystrophy (DMD). The deltaE50-MD dog model of DMD harbors a mutation corresponding to a mutational “hotspot” in the human DMD gene. We used adeno-associated viruses to deliver CRISPR gene editing components to four dogs and examined dystrophin protein expression 6 weeks after intramuscular delivery (n = 2) or 8 weeks after systemic delivery (n = 2). After systemic delivery in skeletal muscle, dystrophin was restored to levels ranging from 3 to 90% of normal, depending on muscle type. In cardiac muscle, dystrophin levels in the dog receiving the highest dose reached 92% of normal. The treated dogs also showed improved muscle histology. These large-animal data support the concept that, with further development, gene editing approaches may prove clinically useful for the treatment of DMD.

scholarly journals Pathological Alterations in the Gastrointestinal Tract of a Porcine Model of DMD

2021 ◽  
Author(s):  
Xiaodong Zhou ◽  
Hongsheng Ouyang ◽  
Daxin Pang ◽  
Renzhi Han ◽  
Xiaochun Tang
Keyword(s):  
Skeletal Muscle ◽  
Gastrointestinal Tract ◽  
Duchenne Muscular Dystrophy ◽  
Muscular Dystrophy ◽  
Nuclear Transfer ◽  
Porcine Model ◽  
Premature Death ◽  
Absorption Capacity ◽  
Gastrointestinal Dysfunction ◽  
Dystrophin Expression

Abstract Patients with Duchenne muscular dystrophy (DMD) develop severe skeletal and cardiac muscle pathologies, which result in premature death. Therefore, the current therapeutic efforts are mainly targeted to correct dystrophin expression in skeletal muscle and heart. However, it was reported that DMD patients may also exhibit gastrointestinal and nutritional problems. How the pathological alterations in gastrointestinal tissues may contribute to the disease are not fully explored. Here we employed the CRISPR/Cas9 system combined with somatic nuclear transfer technology (SCNT) to establish a porcine model of DMD and explored their pathological alterations. We found that genetic disruption of dystrophin expression led1 to morphological gastrointestinal tract alterations, weakened the gastrointestinal tract digestion and absorption capacity, and eventually led to malnutrition and gastric dysfunction in the DMD pigs. This work provides important insights into the pathogenesis of DMD and highlights the need to consider the gastrointestinal dysfunction as an additional therapeutic target for DMD patients.

scholarly journals Restoring Dystrophin Expression in Duchenne Muscular Dystrophy: Current Status of Therapeutic Approaches

2019 ◽  
Vol 9 (1) ◽  
pp. 1 ◽  
Author(s):  
Yuko Shimizu-Motohashi ◽  
Hirofumi Komaki ◽  
Norio Motohashi ◽  
Shin’ichi Takeda ◽  
Toshifumi Yokota ◽  
...  
Keyword(s):  
Gene Therapy ◽  
Duchenne Muscular Dystrophy ◽  
Muscular Dystrophy ◽  
Stop Codon ◽  
Genetic Disorder ◽  
Premature Stop Codon ◽  
Exon Skipping ◽  
Current Status ◽  
Therapeutic Approaches ◽  
Dystrophin Expression

Duchenne muscular dystrophy (DMD), a rare genetic disorder characterized by progressive muscle weakness, is caused by the absence or a decreased amount of the muscle cytoskeletal protein dystrophin. Currently, several therapeutic approaches to cure DMD are being investigated, which can be categorized into two groups: therapies that aim to restore dystrophin expression, and those that aim to compensate for the lack of dystrophin. Therapies that restore dystrophin expression include read-through therapy, exon skipping, vector-mediated gene therapy, and cell therapy. Of these approaches, the most advanced are the read-through and exon skipping therapies. In 2014, ataluren, a drug that can promote ribosomal read-through of mRNA containing a premature stop codon, was conditionally approved in Europe. In 2016, eteplirsen, a morpholino-based chemical capable of skipping exon 51 in premature mRNA, received conditional approval in the USA. Clinical trials on vector-mediated gene therapy carrying micro- and mini- dystrophin are underway. More innovative therapeutic approaches include CRISPR/Cas9-based genome editing and stem cell-based cell therapies. Here we review the current status of therapeutic approaches for DMD, focusing on therapeutic approaches that can restore dystrophin.

scholarly journals 504. Restoration of Dystrophin Expression by Gene Editing with S. aureus Cas9 in Models of Duchenne Muscular Dystrophy

2016 ◽  
Vol 24 ◽  
pp. S201 ◽  
Author(s):  
Jacqueline N. Robinson-Hamm ◽  
Christopher E. Nelson ◽  
Ruth M. Castellanos Rivera ◽  
Annemieke Aartsma-Rus ◽  
Aravind Asokan ◽  
...  
Keyword(s):  
Duchenne Muscular Dystrophy ◽  
Muscular Dystrophy ◽  
Gene Editing ◽  
Dystrophin Expression

scholarly journals Dystrophin and calcium current are decreased in cardiomyocytes expressing Cre enzyme driven by αMHC but not TNT promoter

2019 ◽  
Vol 9 (1) ◽  
Author(s):  
Ludovic Gillet ◽  
Sabrina Guichard ◽  
Maria C. Essers ◽  
Jean-Sébastien Rougier ◽  
Hugues Abriel
Keyword(s):  
Calcium Current ◽  
Troponin T ◽  
Cre Recombinase ◽  
Mrna Levels ◽  
Gene Encoding ◽  
Cardiac Phenotype ◽  
Dystrophin Expression ◽  
Mouse Tissues ◽  
And Function ◽  
Experimental Use

AbstractThe Cre/lox system is a potent technology to control gene expression in mouse tissues. However, cardiac-specific Cre recombinase expression alone can lead to cardiac alterations when no loxP sites are present, which is not well understood. Many loxP-like sites have been identified in the mouse genome that might be Cre sensitive. One of them is located in the Dmd gene encoding dystrophin, a protein important for the function and stabilization of voltage-gated calcium (Cav1.2) and sodium (Nav1.5) channels, respectively. Here, we investigate whether Cre affects dystrophin expression and function in hearts without loxP sites in the genome. In mice expressing Cre under the alpha-myosin heavy chain (MHC-Cre) or Troponin T (TNT-Cre) promoter, we investigated dystrophin expression, Nav1.5 expression, and Cav1.2 function. Compared to age-matched MHC-Cre− mice, dystrophin protein level was significantly decreased in hearts from MHC-Cre+ mice of more than 12-weeks-old. Quantitative RT-PCR revealed decreased mRNA levels of Dmd gene. Unexpectedly, calcium current (ICaL), but not Nav1.5 protein expression was altered in those mice. Surprisingly, in hearts from 12-week-old and older TNT-Cre+ mice, neither ICaL nor dystrophin and Nav1.5 protein content were altered compared to TNT-Cre−. Cre recombinase unpredictably affects cardiac phenotype, and Cre-expressing mouse models should be carefully investigated before experimental use.

Investigating the pathology of Emery–Dreifuss muscular dystrophy

2008 ◽  
Vol 36 (6) ◽  
pp. 1335-1338 ◽  
Author(s):  
Susan C. Brown ◽  
Richard J. Piercy ◽  
Francesco Muntoni ◽  
Caroline A. Sewry
Keyword(s):  
Muscular Dystrophy ◽  
Animal Models ◽  
Genetic Background ◽  
Autosomal Dominant ◽  
Genetic Disorder ◽  
Clinical Severity ◽  
Lamin A ◽  
Pathological Findings ◽  
Gene Encoding ◽  
Insight Into

EDMD (Emery–Dreifuss muscular dystrophy) is caused by mutations in either the gene encoding for lamin A/C (LMNA) located at 1q21.2–q21.3 or emerin (EMD) located at Xq28. Autosomal dominant EDMD caused by LMNA mutations is more common than the X-linked form and often more severe, with an earlier onset. At the histological and histochemical levels, both X-linked and autosomal dominant EDMD appear similar. However, individuals with the same genetic disorder often show remarkable differences in clinical severity, a finding generally attributed to the genetic background. The clinical and pathological findings in EDMD patients found to have mutations in more than one gene are also discussed. There is now much interest in the phenotype of several animal models for EDMD which should lead to an increased insight into the pathogenesis of this disorder, particularly that relating to the heart phenotype.

scholarly journals Pathological alterations in the gastrointestinal tract of a porcine model of DMD

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Xiaodong Zou ◽  
Hongsheng Ouyang ◽  
Daxin Pang ◽  
Renzhi Han ◽  
Xiaochun Tang
Keyword(s):  
Skeletal Muscle ◽  
Gastrointestinal Tract ◽  
Duchenne Muscular Dystrophy ◽  
Muscular Dystrophy ◽  
Nuclear Transfer ◽  
Porcine Model ◽  
Premature Death ◽  
Absorption Capacity ◽  
Gastrointestinal Dysfunction ◽  
Dystrophin Expression

Abstract Background Patients with Duchenne muscular dystrophy (DMD) develop severe skeletal and cardiac muscle pathologies, which result in premature death. Therefore, the current therapeutic efforts are mainly targeted to correct dystrophin expression in skeletal muscle and heart. However, it was reported that DMD patients may also exhibit gastrointestinal and nutritional problems. How the pathological alterations in gastrointestinal tissues contribute to the disease are not fully explored. Results Here we employed the CRISPR/Cas9 system combined with somatic nuclear transfer technology (SCNT) to establish a porcine model of DMD and explored their pathological alterations. We found that genetic disruption of dystrophin expression led to morphological gastrointestinal tract alterations, weakened the gastrointestinal tract digestion and absorption capacity, and eventually led to malnutrition and gastric dysfunction in the DMD pigs. Conclusions This work provides important insights into the pathogenesis of DMD and highlights the need to consider the gastrointestinal dysfunction as an additional therapeutic target for DMD patients.

scholarly journals Effects of systemic multiexon skipping with peptide-conjugated morpholinos in the heart of a dog model of Duchenne muscular dystrophy

2017 ◽  
Vol 114 (16) ◽  
pp. 4213-4218 ◽  
Author(s):  
Yusuke Echigoya ◽  
Akinori Nakamura ◽  
Tetsuya Nagata ◽  
Nobuyuki Urasawa ◽  
Kenji Rowel Q. Lim ◽  
...  
Keyword(s):  
Duchenne Muscular Dystrophy ◽  
Muscular Dystrophy ◽  
Cardiac Muscle ◽  
Genetic Disorder ◽  
Cardiac Conduction ◽  
Purkinje Fibers ◽  
Dystrophin Expression ◽  
Dog Model ◽  
Cardiac Purkinje Fibers ◽  
Systemic Treatments

Duchenne muscular dystrophy (DMD) is a lethal genetic disorder caused by an absence of the dystrophin protein in bodywide muscles, including the heart. Cardiomyopathy is a leading cause of death in DMD. Exon skipping via synthetic phosphorodiamidate morpholino oligomers (PMOs) represents one of the most promising therapeutic options, yet PMOs have shown very little efficacy in cardiac muscle. To increase therapeutic potency in cardiac muscle, we tested a next-generation morpholino: arginine-rich, cell-penetrating peptide-conjugated PMOs (PPMOs) in the canine X-linked muscular dystrophy in Japan (CXMDJ) dog model of DMD. A PPMO cocktail designed to skip dystrophin exons 6 and 8 was injected intramuscularly, intracoronarily, or intravenously into CXMDJ dogs. Intravenous injections with PPMOs restored dystrophin expression in the myocardium and cardiac Purkinje fibers, as well as skeletal muscles. Vacuole degeneration of cardiac Purkinje fibers, as seen in DMD patients, was ameliorated in PPMO-treated dogs. Although symptoms and functions in skeletal muscle were not ameliorated by i.v. treatment, electrocardiogram abnormalities (increased Q-amplitude and Q/R ratio) were improved in CXMDJ dogs after intracoronary or i.v. administration. No obvious evidence of toxicity was found in blood tests throughout the monitoring period of one or four systemic treatments with the PPMO cocktail (12 mg/kg/injection). The present study reports the rescue of dystrophin expression and recovery of the conduction system in the heart of dystrophic dogs by PPMO-mediated multiexon skipping. We demonstrate that rescued dystrophin expression in the Purkinje fibers leads to the improvement/prevention of cardiac conduction abnormalities in the dystrophic heart.

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