AAV CRISPR editing rescues cardiac and muscle function for 18 months in dystrophic mice

Hakim, Chady H.; Wasala, Nalinda B.; Nelson, Christopher E.; Wasala, Lakmini P.; Yue, Yongping; Louderman, Jacqueline A.; Lessa, Thais Borges; Dai, Aihua; Zhang, Keqing; Jenkins, Gregory J.; Nance, Michael E.; Pan, Xiufang; Kodippili, Kasun; Yang, N. Nora; Chen, Shijie; Gersbach, Charles A.; Duan, Dongsheng

doi:10.1172/jci.insight.124297

Cited by 81 publications

(88 citation statements)

References 39 publications

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“…The percentage of dystrophin-positive fibers in the heart strikingly rose from 10% at 1:1 Cas9:gRNA vector to 94% at 1:10 Cas9:gRNA vector, on average. This corroborated results from an earlier study by Hakim et al (2018), which found that the gRNA vector genome was preferentially depleted in vivo compared to the Cas9 vector genome [109]. In that study, the authors similarly demonstrated that increasing the amount of gRNA vector provided to mdx mice, at a 1:3 Cas9:gRNA vector ratio, led to increased dystrophin rescue in the heart post-treatment.…”

Section: Studies Using Animal Modelssupporting

confidence: 86%

“…Assessment of therapeutic success was performed from 12 to 19 months post-injection. The restoration of dystrophin in the heart was persistent across the three, with up to 20% dystrophin of healthy levels observed at 18 months post-treatment in one study by Western blotting [109]. Dystrophin-positive fibers were observed in all studies by IHC.…”

Section: Studies Using Animal Modelsmentioning

confidence: 81%

“…Preprints (www.preprints.org) | NOT PEER-REVIEWED | Posted: 29 December 2019 doi:10.20944/preprints201912.0385.v1 All but one of the studies in Table 2 were done using mouse models, with the majority on correcting the genetic defect in mdx mice [100,103,[106][107][108][109][110][111]. It is difficult to make comparisons of therapeutic efficacy across studies, not only because of all the variations in experimental design and genome editing strategy but also because not all of these studies provided quantitative results.…”

Section: Studies Using Animal Modelsmentioning

confidence: 99%

“…In that study, the authors similarly demonstrated that increasing the amount of gRNA vector provided to mdx mice, at a 1:3 Cas9:gRNA vector ratio, led to increased dystrophin rescue in the heart post-treatment. It is surmised that the limiting property of gRNAs is related to how the gRNA vector adopts an unstable hairpin/cruciform structure in solution, how gRNAs are critical in helping stabilize the conformation of Cas9 to its active form, how fast gRNA turnover is, or how increasing gRNA vector dose ensures that more nuclei express gRNA and are thus amenable to being acted upon by Cas9 [109,113]. Whichever the reason, further investigating this relationship may lead to substantial improvements in the development of CRISPR therapies.…”

Section: Studies Using Animal Modelsmentioning

confidence: 99%

“…Recently, in response to the above limitations, three studies were published evaluating the long-term therapeutic efficacy of DMD CRISPR treatment in mdx mice [109][110][111]. All aimed to delete exon 23 from the Dmd gene, either by itself or along with exons 21 and 22, producing an in-frame transcript for dystrophin translation.…”

Section: Studies Using Animal Modelsmentioning

confidence: 99%

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Genome Editing for the Understanding and Treatment of Cardiomyopathies

Nguyen¹,

Lim²,

Yokota³

2019

Preprint

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Cardiomyopathies are diseases of heart muscle, a significant percentage of which are genetic in origin. Cardiomyopathies can be classified as dilated, hypertrophic, restrictive, arrhythmogenic right ventricular or left ventricular non-compaction, although mixed morphologies are possible. A subset of neuromuscular disorders, notably Duchenne and Becker muscular dystrophies, are also characterized by cardiomyopathy aside from skeletal myopathy. The global burden of cardiomyopathies is certainly high, necessitating further research and novel therapies. Genome editing tools, which include zinc finger nucleases (ZFNs), transcription activator-like effector nucleases (TALENs) and clustered regularly interspaced short palindromic repeats (CRISPR) systems have emerged as increasingly important technologies in studying this group of cardiovascular disorders. In this review, we discuss the applications of genome editing in the understanding and treatment of cardiomyopathy. We also describe recent advances in genome editing that may help improve these applications, and some future prospects for genome editing in cardiomyopathy treatment.Keywords: dilated cardiomyopathy (DCM); hypertrophic cardiomyopathy (HCM); restrictive cardiomyopathy (RCM); arrhythmogenic right ventricular cardiomyopathy (ARVC); left ventricular non-compaction cardiomyopathy (LVNC); Duchenne muscular dystrophy; dystrophin; genome editing; CRISPR/Cas9; Cpf1 (Cas12a) Cardiomyopathy is also a major manifestation in a subset of neuromuscular disorders [7,8]. Cardiomyopathy is most commonly associated with Duchenne muscular dystrophy (DMD) and Becker muscular dystrophy (BMD), both X-linked recessive disorders caused by mutations in the dystrophin (DMD) gene [8]. DMD codes for dystrophin, a cytoskeletal protein that maintains the Preprints (www.preprints.org) | NOT PEER-REVIEWED | Posted: structural integrity of muscle fibres during contraction-relaxation cycles. Loss-of-function mutations in the DMD gene result in an absence of dystrophin, leading to DMD [9][10][11]. Mutations maintaining the DMD reading frame produce truncated but partly functional dystrophin, and often result in a milder form of the disease known as BMD [12,13]. Cardiomyopathy is a leading cause of death in both DMD and BMD patients and is routinely described as being DCM [13,14]. Other neuromuscular disorders associated with cardiomyopathy include the limb girdle muscle dystrophies, myotonic dystrophy, and Friedreich ataxia [15].Advances in molecular genetics enable genome editing to be incorporated into various fields of biomedical research, including the cardiovascular sciences. Currently, the most commonly used tools are zinc finger nucleases (ZFNs), transcription activator-like effector nucleases (TALENs), and clustered regularly interspaced short palindromic repeats (CRISPR) [16]. Both ZFNs and TALENs are engineered restriction enzymes created by combining a DNA binding domain with a DNA cleavage domain adapted from the FokI restriction enzyme [17][18][19]. The DNA binding domain in Z...

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Section: Studies Using Animal Modelssupporting

confidence: 86%

Section: Studies Using Animal Modelsmentioning

confidence: 81%

Section: Studies Using Animal Modelsmentioning

confidence: 99%

Section: Studies Using Animal Modelsmentioning

confidence: 99%

Section: Studies Using Animal Modelsmentioning

confidence: 99%

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Genome Editing for the Understanding and Treatment of Cardiomyopathies

Nguyen¹,

Lim²,

Yokota³

2019

Preprint

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A potential therapeutic effect of catalpol in Duchenne muscular dystrophy revealed by binding with TAK1

Zhao

Jiang

et al. 2020

J cachexia sarcopenia muscle

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Background Duchenne muscular dystrophy (DMD) is a progressive muscle disease caused by the loss of dystrophin, which results in inflammation, fibrosis, and the inhibition of myoblast differentiation in skeletal muscle. Catalpol, an iridoid glycoside, improves skeletal muscle function by enhancing myogenesis; it has potential to treat DMD. We demonstrate the positive effects of catalpol in dystrophic skeletal muscle. Methods mdx (loss of dystrophin) mice (n = 18 per group) were treated with catalpol (200 mg/kg) for six consecutive weeks. Serum analysis, skeletal muscle performance and histology, muscle contractile function, and gene and protein expression were performed. Molecular docking and ligand-target interactions, RNA interference, immunofluorescence, and plasmids transfection were utilized to explore the protective mechanism in DMD by which catalpol binding with transforming growth factor-βactivated kinase 1 (TAK1) in skeletal muscle. Results Six weeks of catalpol treatment improved whole-body muscle health in mdx mice, which was characterized by reduced plasma creatine kinase (n = 18, À35.1%, P < 0.05) and lactic dehydrogenase (n = 18, À10.3%, P < 0.05) activity. These effects were accompanied by enhanced grip strength (n = 18, +25.4%, P < 0.05) and reduced fibrosis (n = 18, À29.0% for hydroxyproline content, P < 0.05). Moreover, catalpol treatment protected against muscle fatigue and promoted muscle recovery in the tibialis anterior (TA) and diaphragm (DIA) muscles (n = 6, +69.8%, P < 0.05 and + 74.8%, P < 0.001, respectively), which was accompanied by enhanced differentiation in primary myoblasts from DMD patients (n = 6, male, mean age: 4.7 ± 1.9 years) and mdx mice. In addition, catalpol eliminated p-TAK1 overexpression in mdx mice (n = 12, À21.3%, P < 0.05) and primary myoblasts. The catalpol-induced reduction in fibrosis and increased myoblast differentiation resulted from the inhibition of TAK1 phosphorylation, leading to reduced myoblast trans-differentiation into myofibroblasts. Catalpol inhibited the phosphorylation of TAK1 by binding to TAK1, possibly at Asp-206, Thr-208, Asn-211, Glu-297, Lys-294, and Tyr-293. Conclusions Our findings show that catalpol and TAK1 inhibitors substantially improve whole-body muscle health and the function of dystrophic skeletal muscles and may provide a novel therapy for DMD.

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The effect of emerging molecular and genetic therapies on cardiopulmonary disease in Duchenne muscular dystrophy

McKim

Cripe

2020

Pediatric Pulmonology

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Gene therapy is an attractive approach being intensively studied to prevent muscle deterioration in patients with Duchenne muscular dystrophy. While clinical trials are only in early stages, initial reports are promising for its effects on ambulation. Cardiopulmonary failure, however, is the most common cause of mortality in Duchenne muscular dystrophy (DMD) patients, and little is known regarding the prospects for gene therapy on alleviating DMD‐associated cardiomyopathy and respiratory failure. Here we review current knowledge regarding effects of gene therapy on DMD cardiomyopathy and discuss respiratory endpoints that should be considered as outcome measures in future clinical trials.

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AAV CRISPR editing rescues cardiac and muscle function for 18 months in dystrophic mice

Abstract: Conflict of interest: DD is a member of the scientific advisory board of and an equity holder in Solid Biosciences LLC. DD and YY are inventors on patents that were licensed to Solid Biosciences LLC

Cited by 81 publications

References 39 publications

Genome Editing for the Understanding and Treatment of Cardiomyopathies

Genome Editing for the Understanding and Treatment of Cardiomyopathies

A potential therapeutic effect of catalpol in Duchenne muscular dystrophy revealed by binding with TAK1

The effect of emerging molecular and genetic therapies on cardiopulmonary disease in Duchenne muscular dystrophy

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