Mechanical ventilation (MV) is used clinically as a life saving intervention to sustain adequate pulmonary gas exchange in patients that are incapable of maintaining sufficient alveolar ventilation (e.g., patients in respiratory failure). The removal of patients from MV is termed """"""""weaning"""""""" and problems in weaning are extremely common. The inability to wean patients from MV results in increased risk of morbidity and mortality along with higher health care costs. Studies indicate that MV-induced diaphragmatic weakness, due to both atrophy and contractile dysfunction, is a significant contributor to weaning difficulties. Our research reveals that prolonged MV is associated with diaphragmatic oxidative stress and that the activation of two proteases, calpains and caspase-3, is required for MV-induced diaphragmatic atrophy and contractile dysfunction. The mechanism(s) that regulate the activity of these proteases in the diaphragm during prolonged MV are currently unknown and will be addressed in this application. HYPOTHESES:
Aim 1 will test the hypothesis that oxidative stress is essential for activation of both calpain and caspase-3 in the diaphragm during prolonged MV.
Aim 2 will test the postulate that oxidative modification of diaphragmatic myofilament proteins increases their susceptibility to degradation by calpains-1/2 and/or caspase-3.
Aim 3 will test the hypothesis that regulatory cross-talk between calpain and caspase-3 plays a vital role in determining the individual activity of these proteases. APPROACH: We will test these hypotheses using a combination of experimental paradigms including an animal model of MV, peptide mapping, and cell-signaling experiments performed in rat primary skeletal muscle cell culture. Cause and effect will be determined by prevention of MV-induced oxidative stress in the diaphragm and by the independent inhibition of calpains and caspase-3 in primary skeletal muscle myotubes using innovative approaches. SIGNIFICANCE AND LONG-TERM GOAL: Failure to wean patients from MV is an important clinical problem and respiratory muscle weakness is a major contributor to weaning difficulties. Hence, our long-term goal is to develop methods for the prevention of MV-induced diaphragmatic weakness based on an understanding of the cellular mechanisms responsible for MV-induced atrophy in the diaphragm. Moreover, the results of these experiments should provide important information for broader topics such as skeletal muscle wasting due to prolonged bed rest, immobilization, and disease states.

Public Health Relevance

Mechanical ventilation (MV) is used to sustain pulmonary gas exchange in patients that are incapable of maintaining adequate alveolar ventilation;the withdrawal of MV from patients is referred to as """"""""weaning"""""""" and problems in weaning from MV are extremely common. Numerous studies reveal that MV-induced diaphragmatic weakness, due to both atrophy and contractile dysfunction, is an important contributor to weaning difficulties. Our long-term goal is to develop methods for the prevention of MV-induced diaphragmatic weakness based on an understanding of the cellular mechanisms responsible for MV-induced diaphragmatic atrophy and contractile dysfunction.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Project (R01)
Project #
5R01HL087839-04
Application #
8252152
Study Section
Skeletal Muscle and Exercise Physiology Study Section (SMEP)
Program Officer
Harabin, Andrea L
Project Start
2009-06-01
Project End
2014-04-30
Budget Start
2012-05-01
Budget End
2014-04-30
Support Year
4
Fiscal Year
2012
Total Cost
$357,732
Indirect Cost
$110,232
Name
University of Florida
Department
Type
Other Domestic Higher Education
DUNS #
969663814
City
Gainesville
State
FL
Country
United States
Zip Code
32611
Talbert, Erin E; Smuder, Ashley J; Kwon, Oh Sung et al. (2016) Blockage of the Ryanodine Receptor via Azumolene Does Not Prevent Mechanical Ventilation-Induced Diaphragm Atrophy. PLoS One 11:e0148161
Powers, Scott K (2014) Can antioxidants protect against disuse muscle atrophy? Sports Med 44 Suppl 2:S155-65
Smuder, Ashley J; Nelson, W Bradley; Hudson, Matthew B et al. (2014) Inhibition of the ubiquitin-proteasome pathway does not protect against ventilator-induced accelerated proteolysis or atrophy in the diaphragm. Anesthesiology 121:115-26
Roberts, Brandon M; Ahn, Bumsoo; Smuder, Ashley J et al. (2013) Diaphragm and ventilatory dysfunction during cancer cachexia. FASEB J 27:2600-10
Bruells, Christian S; Smuder, Ashley J; Reiss, Lucy K et al. (2013) Negative pressure ventilation and positive pressure ventilation promote comparable levels of ventilator-induced diaphragmatic dysfunction in rats. Anesthesiology 119:652-62
Powers, Scott K; Wiggs, Michael P; Duarte, Jose A et al. (2012) Mitochondrial signaling contributes to disuse muscle atrophy. Am J Physiol Endocrinol Metab 303:E31-9
Powers, Scott K; Smuder, Ashley J; Judge, Andrew R (2012) Oxidative stress and disuse muscle atrophy: cause or consequence? Curr Opin Clin Nutr Metab Care 15:240-5
Davis 3rd, Robert T; Bruells, Christian S; Stabley, John N et al. (2012) Mechanical ventilation reduces rat diaphragm blood flow and impairs oxygen delivery and uptake. Crit Care Med 40:2858-66
Nelson, W Bradley; Smuder, Ashley J; Hudson, Matthew B et al. (2012) Cross-talk between the calpain and caspase-3 proteolytic systems in the diaphragm during prolonged mechanical ventilation. Crit Care Med 40:1857-63
Smuder, Ashley J; Hudson, Matthew B; Nelson, W Bradley et al. (2012) Nuclear factor-κB signaling contributes to mechanical ventilation-induced diaphragm weakness*. Crit Care Med 40:927-34

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