Delivery of Recombinant Adeno-Associated Virus Vectors to Rat Diaphragm Muscle via Direct Intramuscular Injection

SmuderAshley, J; FalkDarin, J; SollanekKurt, J; Nelson, W. Bradley; PowersScott, K

doi:10.1089/hgtb.2013.055

Cited by 13 publications

(21 citation statements)

References 43 publications

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“…To increase the expression of HSP72 in the diaphragm, we delivered an adeno‐associated virus (AAV) vector to the costal diaphragm (AAV‐HSP72) via direct intramuscular injection as previously described . Following this procedure, animals were administered buprenorphine (0.1 mg/kg) prior to awakening.…”

Section: Methodsmentioning

confidence: 99%

“…Following this procedure, animals were administered buprenorphine (0.1 mg/kg) prior to awakening. Additional doses were administered as needed for 72 h. This method of gene transfer effectively transduces the rat diaphragm with no adverse side effects to the muscle . Preliminary experiments were performed to determine the proper concentration of AAV‐HSP72 to increase HSP72 protein expression ~50% above basal (control) levels.…”

Section: Methodsmentioning

confidence: 99%

“…AAV delivery protocol To increase the expression of HSP72 in the diaphragm, we delivered an adeno-associated virus (AAV) vector to the costal diaphragm (AAV-HSP72) via direct intramuscular injection as previously described. 21,22 Following this procedure, animals were administered buprenorphine (0.1 mg/kg) prior to awakening. Additional doses were administered as needed for 72 h. This method of gene transfer effectively transduces the rat diaphragm with no adverse side effects to the muscle.…”

Section: Experiments Twomentioning

confidence: 99%

“…Additional doses were administered as needed for 72 h. This method of gene transfer effectively transduces the rat diaphragm with no adverse side effects to the muscle. 21,22 Preliminary experiments were performed to determine the proper concentration of AAV-HSP72 to increase HSP72 protein expression~50% above basal (control) levels. This percentage increase was chosen to match the increase achieved by our exercise training protocol.…”

Section: Experiments Twomentioning

confidence: 99%

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Effects of exercise preconditioning and HSP72 on diaphragm muscle function during mechanical ventilation

Smuder

Morton

Hall

et al. 2019

J cachexia sarcopenia muscle

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Background Mechanical ventilation (MV) is a life‐saving measure for patients in respiratory failure. However, prolonged MV results in significant diaphragm atrophy and contractile dysfunction, a condition referred to as ventilator‐induced diaphragm dysfunction (VIDD). While there are currently no clinically approved countermeasures to prevent VIDD, increased expression of heat shock protein 72 (HSP72) has been demonstrated to attenuate inactivity‐induced muscle wasting. HSP72 elicits cytoprotection via inhibition of NF‐κB and FoxO transcriptional activity, which contribute to VIDD. In addition, exercise‐induced prevention of VIDD is characterized by an increase in the concentration of HSP72 in the diaphragm. Therefore, we tested the hypothesis that increased HSP72 expression is required for the exercise‐induced prevention of VIDD. We also determined whether increasing the abundance of HSP72 in the diaphragm, independent of exercise, is sufficient to prevent VIDD. Methods Cause and effect was determined by inhibiting the endurance exercise‐induced increase in HSP72 in the diaphragm of exercise trained animals exposed to prolonged MV via administration of an antisense oligonucleotide targeting HSP72. Additional experiments were performed to determine if increasing HSP72 in the diaphragm via genetic (rAAV‐HSP72) or pharmacological (BGP‐15) overexpression is sufficient to prevent VIDD. Results Our results demonstrate that the exercise‐induced increase in HSP72 protein abundance is required for the protective effects of exercise against VIDD. Moreover, both rAAV‐HSP72 and BGP‐15‐induced overexpression of HSP72 were sufficient to prevent VIDD. In addition, modification of HSP72 in the diaphragm is inversely related to the expression of NF‐κB and FoxO target genes. Conclusions HSP72 overexpression in the diaphragm is an effective intervention to prevent MV‐induced oxidative stress and the transcriptional activity of NF‐κB and FoxO. Therefore, overexpression of HSP72 in the diaphragm is a potential therapeutic target to protect against VIDD.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Experiments Twomentioning

confidence: 99%

Section: Experiments Twomentioning

confidence: 99%

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Effects of exercise preconditioning and HSP72 on diaphragm muscle function during mechanical ventilation

Smuder

Morton

Hall

et al. 2019

J cachexia sarcopenia muscle

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“…This innovative technique is described in a recent report (18). Specifically, this method of gene transfer was reported to effectively transduce the rat diaphragm with no adverse side effects (18). …”

Section: Methodsmentioning

confidence: 99%

Inhibition of Forkhead BoxO–Specific Transcription Prevents Mechanical Ventilation–Induced Diaphragm Dysfunction

Smuder

Sollanek

Min

et al. 2015

Critical Care Medicine

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Background Mechanical ventilation (MV) is a life-saving measure for patients in respiratory failure. However, prolonged MV results in diaphragm weakness, which contributes to problems in weaning from the ventilator. Therefore, identifying the signaling pathways responsible for MV-induced diaphragm weakness is essential to developing effective countermeasures to combat this important problem. In this regard, the FOXO family of transcription factors is activated in the diaphragm during MV and FOXO-specific transcription can lead to enhanced proteolysis and muscle protein breakdown. Currently, the role that FOXO activation plays in the development of MV-induced diaphragm weakness remains unknown. Objectives This study tested the hypothesis that MV-induced increases in FOXO signaling contribute to ventilator-induced diaphragm weakness. Interventions Cause and effect was determined by inhibiting the activation of FOXO in the rat diaphragm through the use of a dominant negative FOXO (dnFOXO) adeno-associated virus vector delivered directly to the diaphragm. Measurements and Main Results Our results demonstrate that prolonged (12 hrs) MV results in a significant decrease in both diaphragm muscle fiber size and diaphragm specific force production. However, MV animals treated with dnFOXO showed a significant attenuation of both diaphragm atrophy and contractile dysfunction. In addition, inhibiting FOXO transcription attenuated the MV-induced activation of the ubiquitin-proteasome system, the autophagy/lysosomal system and caspase-3. Conclusions FOXO is necessary for the activation of key proteolytic systems essential for MV-induced diaphragm atrophy and contractile dysfunction. Collectively, these results suggest that targeting FOXO transcription could be a key therapeutic target to combat VIDD.

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Gene therapy and respiratory neuroplasticity

Mantilla

2017

Experimental Neurology

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Delivery of Recombinant Adeno-Associated Virus Vectors to Rat Diaphragm Muscle via Direct Intramuscular Injection

Cited by 13 publications

References 43 publications

Effects of exercise preconditioning and HSP72 on diaphragm muscle function during mechanical ventilation

Effects of exercise preconditioning and HSP72 on diaphragm muscle function during mechanical ventilation

Inhibition of Forkhead BoxO–Specific Transcription Prevents Mechanical Ventilation–Induced Diaphragm Dysfunction

Gene therapy and respiratory neuroplasticity

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