Micro-dystrophin gene therapy demonstrates long-term cardiac efficacy in a severe Duchenne muscular dystrophy model

Piepho, Arden B.; Lowe, Jeovanna; Cumby, Laurel R.; Dorn, Lisa E.; Lake, Dana M.; Rastogi, Neha; Gertzen, Megan D.; Sturgill, Sarah L.; Odom, Guy L.; Ziolo, Mark T.; Accornero, Federica; Chamberlain, Jeffrey S.; Rafael‐Fortney, Jill A.

doi:10.1016/j.omtm.2023.02.001

Cited by 8 publications

(4 citation statements)

References 24 publications

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“…Various injectable hydrogels have been developed in recent years for purposes such as drug delivery and cellular therapy 63 ; therefore, using a hydrogel without components that may coincide with contrast generated from AAV particles themselves would be useful for more distinct detection. The viral load for in vivo hydrogel administration used in this study was approximately 6.67 × 10 10 vg, which is on the lower end of the 10 9 –10 12 vg range used in recent studies evaluating AAV‐mediated gene therapy treatments for muscular pathologies in mice 64–67 . Based on the results of this study, the lower bound of viral load for sufficient detection via CEST imaging is estimated to be approximately 10 6 vg per gram of tissue.…”

Section: Discussionmentioning

confidence: 91%

“…The viral load for in vivo hydrogel administration used in this study was approximately 6.67 × 10 10 vg, which is on the lower end of the 10 9 -10 12 vg range used in recent studies evaluating AAV-mediated gene therapy treatments for muscular pathologies in mice. [64][65][66][67] Based on the results of this study, the lower bound of viral load for sufficient detection via CEST imaging is estimated to be approximately 10 6 vg per gram of tissue. This approximation assumes some loss in sensitivity in tissue with background signal.…”

Section: F I G U R Ementioning

confidence: 95%

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Imaging of adeno‐associated viral capsids for purposes of gene editing using CEST NMR/MRI

Lam,

Velasquez,

Ogiyama

et al. 2024

Magnetic Resonance in Med

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PurposeGene therapy using adeno‐associated virus (AAV) vector‐mediated gene delivery has undergone substantial growth in recent years with promising results in both preclinical and clinical studies, as well as emerging regulatory approval. However, the inability to quantify the efficacy of gene therapy from cellular delivery of gene‐editing technology to specific functional outcomes is an obstacle for efficient development of gene therapy treatments. Building on prior works that used the CEST reporter gene lysine rich protein, we hypothesized that AAV viral capsids may generate endogenous CEST contrast from an abundance of surface lysine residues.MethodsNMR experiments were performed on isolated solutions of AAV serotypes 1–9 on a Bruker 800‐MHz vertical scanner. In vitro experiments were performed for testing of CEST‐NMR contrast of AAV2 capsids under varying pH, density, biological transduction stage, and across multiple serotypes and mixed biological media. Reverse transcriptase–polymerase chain reaction was used to quantify virus concentration. Subsequent experiments at 7 T optimized CEST saturation schemes for AAV contrast detection and detected AAV2 particles encapsulated in a biocompatible hydrogel administered in the hind limb of mice.ResultsCEST‐NMR experiments revealed CEST contrast up to 52% for AAV2 viral capsids between 0.6 and 0.8 ppm. CEST contrast generated by AAV2 demonstrated high levels of CEST contrast across a variety of chemical environments, concentrations, and saturation schemes. AAV2 CEST contrast displayed significant positive correlations with capsid density (R2 > 0.99, p < 0.001), pH (R2 = 0.97, p = 0.01), and viral titer per cell count (R2 = 0.92, p < 0.001). Transition to a preclinical field strength yielded up to 11.8% CEST contrast following optimization of saturation parameters. In vivo detection revealed statistically significant molecular contrast between viral and empty hydrogels using both mean values (4.67 ± 0.75% AAV2 vs. 3.47 ± 0.87% empty hydrogel, p = 0.02) and quantile analysis.ConclusionAAV2 viral capsids exhibit strong capacity as an endogenous CEST contrast agent and can potentially be used for monitoring and evaluation of AAV vector‐mediated gene therapy protocols.

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Section: Discussionmentioning

confidence: 91%

Section: F I G U R Ementioning

confidence: 95%

Imaging of adeno‐associated viral capsids for purposes of gene editing using CEST NMR/MRI

Lam,

Velasquez,

Ogiyama

et al. 2024

Magnetic Resonance in Med

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“…Experiments have tested the efficacy of delivering a small dystrophin fragment called micro‐dystrophin 5 (μDys5). μDys5 has been packaged into an AAV (AAV‐μDys5) and injected into DMD mice at 4 weeks of age, which prevented fibrotic and inflammatory processes in the heart up to 18 months of age (Piepho et al., 2023 ). Micro‐dystrophins are unlikely to fully replace the function of full‐length dystrophin inside a DMD cell, so it may be necessary to increase the amount of full‐length or near full‐length dystrophin to treat DMD.…”

Section: The Practicalities and Hurdles Of Delivering Durable Gene Th...mentioning

confidence: 99%

Targeted genetic therapies for inherited disorders that affect both cardiac and skeletal muscle

Psaras,

Toepfer

2023

Experimental Physiology

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Skeletal myopathies and ataxias with secondary cardiac involvement are complex, progressive and debilitating conditions. As life expectancy increases across these conditions, cardiac involvement often becomes more prominent. This highlights the need for targeted therapies that address these evolving cardiac pathologies. Musculopathies by and large lack cures that directly target the genetic basis of the diseases; however, as our understanding of the genetic causes of these conditions has evolved, it has become tractable to develop targeted therapies using biologics, to design precision approaches to target the primary genetic causes of these varied diseases. Using the examples of Duchenne muscular dystrophy, Friedreich ataxia and Pompe disease, we discuss how the genetic causes of such diseases derail diverse homeostatic, energetic and signalling pathways, which span multiple cellular systems in varied tissues across the body. We outline existing therapeutics and treatments in the context of emerging novel genetic approaches. We discuss the hurdles that the field must overcome to deliver targeted therapies across the many tissue types affected in primary myopathies.

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“…We have investigated an alternative approach to restoring functional dysferlin to skeletal muscle via AAV transduction: creation of smaller versions of the protein, or “nanodysferlins,” that in principle can retain the activities of native dysferlin but that can be packaged in AAV. This approach has been useful in restoring many of the functions of dystrophin in mice and dogs with dystrophinopathies (e.g., 49 , 50 , 51 , 52 , 53 , 54 , 55 , 56 , 57 ). We have shown that some nanodysferlins can support membrane resealing and slow the progression of dysferlinopathy in dysferlin-null mice.…”

Section: Introductionmentioning

confidence: 99%

Nanodysferlins support membrane repair and binding to TRIM72/MG53 but do not localize to t-tubules or stabilize Ca2+ signaling

Muriel,

Lukyanenko,

Kwiatkowski

et al. 2024

Molecular Therapy - Methods & Clinical Development

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Micro-dystrophin gene therapy demonstrates long-term cardiac efficacy in a severe Duchenne muscular dystrophy model

Cited by 8 publications

References 24 publications

Imaging of adeno‐associated viral capsids for purposes of gene editing using CEST NMR/MRI

Imaging of adeno‐associated viral capsids for purposes of gene editing using CEST NMR/MRI

Targeted genetic therapies for inherited disorders that affect both cardiac and skeletal muscle

Nanodysferlins support membrane repair and binding to TRIM72/MG53 but do not localize to t-tubules or stabilize Ca2+ signaling

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