title: Combined inhibition of cell death induced by apoptosis inducing factor and caspases provides additive neuroprotection in experimental traumatic brain injury.
abstract: Neuronal programmed cell death (PCD) contributes to delayed tissue damage after traumatic brain injury (TBI). Both caspase-dependent and caspase-independent mechanisms have been implicated, with the latter including apoptosis inducing factor (AIF). The peptidyl-proplyl isomerase Cyclophilin A (CypA) transports AIF from the cytosol to the nucleus, a key step for AIF-dependent cell death. We compared the effects of single versus combined inhibition of caspase and AIF pathways in a mouse controlled cortical impact (CCI) model, by examining the effects of CypA gene knockout (CypA(-/-)), caspase inhibition with a pan-caspase inhibitor (boc-aspartyl(OMe)-fluoromethylketone, BAF), or combined modulation. TBI caused caspase activation as well as translocation of AIF to the nucleus. Markers of caspase activation including caspase-specific fodrin cleavage fragments and number of FLIVO-positive cells were reduced in BAF-treated CypA(+/+) mice, whereas markers of AIF activation including AIF/H2AX interaction and AIF translocation to the nucleus were attenuated in CypA(-/-) mice. Each single intervention, (CypA(-/-) or BAF-treated CypA(+/+)) reduced the number of apoptotic cells (TUNEL-positive) in the cortex and improved long-term sensorimotor function; CypA(-/-) also attenuated microglial activation after injury. Importantly, BAF-treated CypA(-/-) mice, showed greater effects than either intervention alone on multiple outcomes including: reduction in TUNEL-positive cells, decrease in neuroinflammation, improved motor and cognitive recovery, and attenuation of lesion volume and neuronal loss in the hippocampus. Using two in vitro neuronal cell death models known to induce AIF-mediated PCD, we also showed that neurons from CypA(-/-) animals were protected and that effects were unrelated to caspase activation. These data indicate that AIF-mediated and caspase-dependent pathways contribute independently and in parallel to secondary injury after TBI, and suggest that combined therapeutic strategies directed at multiple PCD pathways may provide superior neuroprotection than those directed at single mechanisms.
authors.affiliation: Center for Shock, Trauma and Anesthesiology Research and Department of Anesthesiology, University of Maryland School of Medicine, Baltimore, MD 21201, USA.
authors.forename: Chun-Shu
authors.initials: CS
authors.lastname: Piao
authors.forename: David J
authors.initials: DJ
authors.lastname: Loane
authors.forename: Bogdan A
authors.initials: BA
authors.lastname: Stoica
authors.forename: Shihong
authors.initials: S
authors.lastname: Li
authors.forename: Marie
authors.initials: M
authors.lastname: Hanscom
authors.forename: Rainier
authors.initials: R
authors.lastname: Cabatbat
authors.forename: Klas
authors.initials: K
authors.lastname: Blomgren
authors.forename: Alan I
authors.initials: AI
authors.lastname: Faden
mesh_terms: D000590:Amino Acid Chloromethyl Ketones / Q000494:pharmacology; D000818:Animals; D051033:Apoptosis Inducing Factor / Q000494:pharmacology*; D015153:Blotting, Western; D001930:Brain Injuries / Q000188:drug therapy* / Q000473:pathology; D020169:Caspases / Q000494:pharmacology*; D016923:Cell Death / Q000502:physiology*; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D003071:Cognition / Q000187:drug effects; D021984:Cyclophilin A / Q000235:genetics / Q000502:physiology; D015853:Cysteine Proteinase Inhibitors / Q000494:pharmacology; D006624:Hippocampus / Q000473:pathology; D007150:Immunohistochemistry; D047468:Immunoprecipitation; D020287:In Situ Nick-End Labeling; D007276:Injections, Intraventricular; D008279:Magnetic Resonance Imaging; D051379:Mice; D018345:Mice, Knockout; D009068:Movement / Q000187:drug effects; D009474:Neurons / Q000473:pathology; D018696:Neuroprotective Agents; D015398:Signal Transduction / Q000187:drug effects
chemical_list: D000590:Amino Acid Chloromethyl Ketones; D051033:Apoptosis Inducing Factor; D015853:Cysteine Proteinase Inhibitors; D018696:Neuroprotective Agents; C103192:butyloxycarbonyl-O-methyl-aspartyl-fluoromethyl ketone; D020169:Caspases; D021984:Cyclophilin A