AAV1.NT-3 Gene Therapy for Charcot–Marie–Tooth Neuropathy

Sahenk, Zarife; Galloway, Gloria; Clark, K. Reed; Malik, Vinod; Rodino‐Klapac, Louise R.; Kaspar, Brian K.; Chen, Lei; Braganza, Cilwyn; Montgomery, Chrystal L.; Mendell, Jerry R.

doi:10.1038/mt.2013.250

Cited by 87 publications

(120 citation statements)

References 48 publications

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“…5,6 AAV serotype 1 (AAV1) is used for neuroscience applications such as opotogenetics, 7,8 pharmacogenetics, [9][10][11] and gene silencing. 12 AAV1 is also being used in human clinical trials for heart disease, 13 alpha-1 antitrypsin deficiency, 14 limbgirdle muscular dystrophy type 2D, 15 CharcotMarie-Tooth neuropathy, 16 and Becker muscular dystrophy. 17 Given the prevalence of usage and breadth of applications, establishing practical procedures in the research laboratory setting for inactivating AAV is essential for the safe use of AAV and reducing cross-contamination of instruments and equipment.…”

Section: Adeno-associated Virus (Aav)-based Viral Vectorsmentioning

confidence: 99%

Assaying the Stability and Inactivation of AAV Serotype 1 Vectors

Howard

Harvey

2017

Human Gene Therapy Methods

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Adeno-associated virus (AAV) vectors are a commonplace tool for gene delivery ranging from cell culture to human gene therapy. One feature that makes AAV a desirable vector is its stability, in regard to both the duration of transgene expression and retention of infectivity as a viral particle. This study examined the stability of AAV serotype 1 (AAV1) vectors under different conditions. First, transducibility after storage at 4°C decreased 20% over 7 weeks. Over 10 freeze-thaw cycles, the resulting transduction efficiency became variable at 60-120% of a single thaw. Using small stainless steel slugs to mimic a biosafety cabinet or metal lab bench surface, it was found that an AAV1 vector can be reconstituted after 6 days of storage at room temperature. The stability of AAV is a desired feature, but effective decontamination procedures must be available for safety and experimental integrity. Multiple disinfectants commonly used in the laboratory for ability to inactivate an AAV1 vector were tested, and it was found that autoclaving, 0.25% peracetic acid, iodine, or 10% Clorox bleach completely prevented AAV-mediated transgene expression. These data suggest that peracetic acid should be used for inactivating AAV1 vectors on metal-based surfaces or instruments in order to avoid inadvertent transgene expression in human cells or cross-contamination of instruments.

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Section: Adeno-associated Virus (Aav)-based Viral Vectorsmentioning

confidence: 99%

Assaying the Stability and Inactivation of AAV Serotype 1 Vectors

Howard

Harvey

2017

Human Gene Therapy Methods

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“…Regeneration in a mixed culture after NT-3 was locally injected into muscle cells in vitro was intensified, thus preventing the effects of muscle atrophy. Sahenk et al [18] proved that NT-3 is abundant in skeletal muscles. Local endogenous injection can enhance RNA expression in advance, stimulate proliferation of myoblasts, promote the formation of muscle tube fuse into muscle fibers, and accelerate the repair process in the skeletal muscle.…”

Section: Discussionmentioning

confidence: 98%

Effects of Neurotrophin-3 Plasmids on Myocyte Apoptosis and Ca2+-ATPase Content in the Muscle After Nerve Injury in Rats

et al. 2015

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We estimated the influence of plasmids with DNA carrying the neurotrophin-3 (NT-3) gene on apoptosis in the gastrocnemius muscle and content of Ca 2+ -ATPase in the latter after sciatic nerve injury. Sixty adult Wistar rats were randomly divided into the saline (control) and NT-3 groups. The related indices, such as expression of caspase-3 protein, the rate of apoptosis in the muscle evaluated using a TUNEL technique, and the level of Ca 2+ -ATPase estimated using Western blot, were observed. Expression of caspase-3 protein was elevated at different post-operative times after peripheral nerve injury; NT-3 expression and the rate of muscle cell apoptosis decreased, whereas the level of Ca 2+ -ATPase in the sarcoplasmic reticulum increased; significant differences were observed compared with the saline group (P < 0.05). The mitigation mechanism of NT-3 on muscle atrophy after peripheral nerve injury is expressed as inhibition of caspase-3 gene expression, increase in the Ca 2+ -ATPase level, and reduction in the rate of muscle apoptosis.

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“…98 Gene therapy can be considered for replacing the defective genes responsible for recessive CMT types or to express protective neurotrophic molecules. 99,100 The optimal selective gene delivery system has not yet been discovered. Other geneticsbased therapies, such as molecules that can correct nonsense mutations or facilitate gene expression regulations, hold promise in specific therapies for CMT.…”

Section: Summary and Future Directionsmentioning

confidence: 99%