Despite numerous positive animal studies, clinical neuroprotection trials after traumatic brain injury (TBI) have uniformly failed with regard to primary end points and general populations. However, most such therapies have been directed toward a single proposed injury pathway and have targeted very early biochemical changes. Our prior work has shown that cell cycle activation is a key component of secondary injury following TBI;based upon our research and that from other groups, we hypothesize that E2F 1, 2, and 3 are involved in both cell cycle-related neuronal death (CRND) and microglial activation after brain trauma. In the present project we propose to use highly specific E2F decoy oligonucleotides (ODN) to demonstrate the neuroprotective effect of blocking the E2Fs. We will generate cell specific and inducible E2F 1, 2, and 3 knockouts and Prohibitin-1 knock-in to demonstrate that cell cycle activation in neurons and microglia represent separate and additive secondary injury mechanisms. We will also use the retinoblastoma modulator RRD-251 to confirm the neuroprotective effect of blocking the E2Fs. Moreover, we propose to expand our focus beyond the acute injury and test the hypothesis that delayed cell cycle activation contributes to chronic neurodegeneration and microglial activation months after TBI.
Specific aims are to show that: (1) TBI-induced activation of E2F transcription factors is a critical event contributing to acute neuronal cell death and microglial/astrocyte reactivity, and that treatment with highly specific E2F decoy oligonucleotide (ODN) sequences or the Rb modulator RRD-251 attenuates these cellular changes and limits injury-induced pathobiology (2) E2F 1-3 are important contributing initiators of neuronal cell death and activation of microglia, with attenuation of E2F activity in neurons, and microglia showing additive neuroprotective effects;conditional and cell specific triple knockouts (E2F 1-3) will be generated by mating E2F 1-3 loxP mice with our inducible neuronal and microglia specific Cre mice;conditional and cell specific Prohibitin-1 knock-in will be generated by mating Prohibitin-1 loxP mice with our inducible neuronal and microglia specific Cre mice (3) Cell cycle activation after TBI persists beyond the acute period and leads to progressively increased polyploidy of certain neurons in selectively vulnerable brain regions as demonstrated by slide-based cytometry and fluorescence in- situ hybridization (FISH);these changes are not followed by rapid neuronal death but rather serve to predispose to chronic progressive neurodegeneration (4) Induction of conditional, neuron specific or microglial specific triple knockouts (E2F 1-3), or treatment with a the pan-CDK inhibitor CR8, at 1 month after trauma, reduce progressive neuronal hyperploidy, and attenuate chronic progressive neuronal loss and associated neurological impairment detected at 1 year after trauma.

Public Health Relevance

As clinical trials of neuroprotective drugs to treat traumatic brain injury (TBI) have failed to date, there is a critical need to identify new therapeutic strategies for this common and costly disorder. We have previously shown that cell cycle activation represents a therapeutically relevant mechanism responsible for neuronal cell death and microglial activation. Here we propose to continue and extend our work and use transgenic models and a drug-based approach to demonstrate the key role of E2F 1, 2, and 3 as inducers of cell cycle activation following TBI. In addition, we will focus not only on acute injury but als on chronic neurodegeneration following TBI to test our hypothesis that chronic cell cycle activation after TBI is an important contributing factor for trauma-induced neurodegeneration.

Agency
National Institute of Health (NIH)
Institute
National Institute of Neurological Disorders and Stroke (NINDS)
Type
Research Project (R01)
Project #
5R01NS052568-07
Application #
8461123
Study Section
Brain Injury and Neurovascular Pathologies Study Section (BINP)
Program Officer
Hicks, Ramona R
Project Start
2005-07-01
Project End
2017-01-31
Budget Start
2013-02-01
Budget End
2014-01-31
Support Year
7
Fiscal Year
2013
Total Cost
$443,158
Indirect Cost
$154,456
Name
University of Maryland Baltimore
Department
Anesthesiology
Type
Schools of Medicine
DUNS #
188435911
City
Baltimore
State
MD
Country
United States
Zip Code
21201
Faden, Alan I; Loane, David J (2015) Chronic neurodegeneration after traumatic brain injury: Alzheimer disease, chronic traumatic encephalopathy, or persistent neuroinflammation? Neurotherapeutics 12:143-50
Kabadi, Shruti V; Stoica, Bogdan A; Loane, David J et al. (2014) CR8, a novel inhibitor of CDK, limits microglial activation, astrocytosis, neuronal loss, and neurologic dysfunction after experimental traumatic brain injury. J Cereb Blood Flow Metab 34:502-13
Aungst, Stephanie L; Kabadi, Shruti V; Thompson, Scott M et al. (2014) Repeated mild traumatic brain injury causes chronic neuroinflammation, changes in hippocampal synaptic plasticity, and associated cognitive deficits. J Cereb Blood Flow Metab 34:1223-32
Loane, David J; Kumar, Alok; Stoica, Bogdan A et al. (2014) Progressive neurodegeneration after experimental brain trauma: association with chronic microglial activation. J Neuropathol Exp Neurol 73:14-29
Loane, David J; Stoica, Bogdan A; Byrnes, Kimberly R et al. (2013) Activation of mGluR5 and inhibition of NADPH oxidase improves functional recovery after traumatic brain injury. J Neurotrauma 30:403-12
Kabadi, Shruti V; Stoica, Bogdan A; Hanscom, Marie et al. (2012) CR8, a selective and potent CDK inhibitor, provides neuroprotection in experimental traumatic brain injury. Neurotherapeutics 9:405-21
Kabadi, Shruti V; Stoica, Bogdan A; Byrnes, Kimberly R et al. (2012) Selective CDK inhibitor limits neuroinflammation and progressive neurodegeneration after brain trauma. J Cereb Blood Flow Metab 32:137-49
Kabadi, Shruti V; Stoica, Bogdan A; Loane, David J et al. (2012) Cyclin D1 gene ablation confers neuroprotection in traumatic brain injury. J Neurotrauma 29:813-27
Kabadi, Shruti V; Hilton, Genell D; Stoica, Bogdan A et al. (2010) Fluid-percussion-induced traumatic brain injury model in rats. Nat Protoc 5:1552-63
Stoica, Bogdan A; Faden, Alan I (2010) Cell death mechanisms and modulation in traumatic brain injury. Neurotherapeutics 7:3-12

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