Vanessa Mazandi, MD
Childrens Hospital of Philadelphia
Philadelphia, Pennsylvania
Disclosure information not submitted.
David Jang, MD, MSc
Assistant Professor
University of Pennsylvania, United States
Disclosure information not submitted.
Samuel Shin, MD, PhD
Neurocritical Care Fellow
Perelman School of Medicine, University of Pennsylvania, United States
Disclosure information not submitted.
Abhay Ranganathan, BA, MS
Program Coordinator
Children's Hospital of Philadelphia, United States
Disclosure information not submitted.
Nile Delso, MS
Research Assistant
Childrens Hospital of Phialdelphia, United States
Disclosure information not submitted.
William Landis, BS
Research Assistant
Childrens Hospital of Philadelphia, United States
Disclosure information not submitted.
Jonathan Starr, BS
Lab Operations Coordinator
Children's Hospital of Philadelphia, United States
Disclosure information not submitted.
Todd Kilbaugh, MD
Associate Professor of Anesthesia, Critical Care, & Pediatrics
Childrens Hospital of Philadelphia, United States
Disclosure information not submitted.
Title: Effects on mitochondrial dynamics following therapeutic intervention in large animal TBI model
Introduction: Traumatic brain injury [TBI] accounts for a significant source of morbidity and mortality in the United States, affecting 2.8 million people per year. Lifetime costs following TBI have been estimated as high as $76.5 billion dollars. Despite the high costs to society, there are no approved therapeutics. One significant hurdle is the lack of translational large animal platforms in TBI. We examined mitochondrial dynamics - fusion and fission - following intervention with a novel alpha-lipoic acid analogue, CMX-2043, that reduces mitochondrial reactive oxygen species [ROS]. Fusion is thought to be protective, associated with mitochondrial repair and improvement in oxidative phosphorylation following injury. Prolonged stress can lead to fission, causing mitochondrial fragmentation, increased ROS and can trigger apoptosis.
Methods: Yorkshire piglets, n=21, 4-weeks old, were randomized into sham [5], placebo [8] and treatment [8] groups. All groups were treatment blinded. Sham animals had surgical incision similar to injured animals. Placebo and treatment pigs received a moderate-severe controlled cortical impact [CCI] injury. Five days after injury, animals were sacrificed. Brain tissue frozen at -80 degrees Celcius was homogenized for western blot analysis to quantify the proteins involved with fusion (OPA1) and fission (DRP1). Antibody application was done via the iBind method. Antibodies were revealed with SuperSignalTM West Pico PLUS Chemiluminescent Substrate and visualized on iBrightCL1000 by Invitrogen. Mean values for each group were compared using Tukey's Multiple Comparisons test.
Results: DRP1 expression was significantly decreased in CMX-2043 treated subjects as compared to placebo (mean difference: 10329; CI 5143-11513, p< 0.0001). OPA1 expression was not significantly different between CMX-2043 treated subjects and placebo (mean difference: 2993; CI-11867-17854; p=0.8794).
Conclusions: Subjects treated with an ALA analogue, CMX-2043, had significantly lower expression of DRP1, fission, 5 days post-TBI. There was no significant difference in OPA1 expression, fusion, at the same time-point, signaling that fusion activity did not appear to be affected by CMX-2043. Further investigation is required to better characterize the influences and tendencies of mitochondrial dynamics following injury.