Brain derived neurotrophic factor (BDNF), a critical signaling molecule in the brain, is functionally linked to essential cellular processes, such as those associated with learning and memory. Altered BDNF signaling is thought to play a crucial role in dysregulated neuroplasticity underlying multiple neurological diseases, including epilepsy, yet the complex molecular determinants of these important brain processes are not fully understood. Our laboratories discovered that BDNF modulates inhibition, in part, through activation of the Janus Kinase (JAK)/Signal Transducer and Activator of Transcription (STAT) signaling pathway. Employing primary cultured neurons and rodent models, we have reported that BDNF-induced JAK/STAT signaling represses the expression of synaptic ?1 containing GABAA receptors (GABARs) following epileptogenic brain injuries, including status epilepticus (SE) and brain trauma (TBI), and that JAK/STAT inhibition at the time of injury can reduce the severity of subsequent epilepsy. We also have preliminary evidence that BDNF-induced JAK/STAT signaling is mediated by TrKB activation at the cell surface promoting JAK2 autophosphorylation within an intracellular signalsome containing p75 neurotrophin receptors (a complex referred to as (i)p75NTRJ), leading to STAT3 recruitment and activation. Subsequent transport of the (i)p75NTRJ signalsome into the nucleus may alter transcription of multiple target genes, including inducible early cAMP repressor (ICER) that binds to and represses the ?1 GABAR gene after SE. To test our hypothesis that BDNF-induced JAK/STAT activation is dysregulated following brain injury causing a systematic change in the transcription of multiple genes that promote epileptogenesis, we will use an unbiased transcriptomic approach, including RNA-seq and ChIP-seq with antibodies to pSTAT3, p75NTR, JAK2, and ICER, in a preclinical epilepsy model utilizing wild-type and transgenic mice with inducible deletion of p75NTR or STAT3 genes. Specifically, we will: 1) Identify the target genes of (i)p75NTRJ that change their expression after epileptogenic brain injury and whether they are specific to neurons or part of a general cellular response using neuron selective or global p75NTR or STAT3 deletion/inhibition; 2) Mechanistically test the relationship of target genes identified in Aim 1 to the genomic response of individual neurons treated with BDNF or kainate; and 3) Determine the functional consequences of neuronal selective or global p75NTR or STAT3 removal on epileptogenesis and cognitive co-morbidities following brain injury. Combining transcriptomic analysis with mouse genetics in an animal model of epilepsy, we will expand our current knowledge of GABAR gene regulation after brain injury to appreciate the complex patterns of genomic changes that underlie epileptogenesis. Such understanding is essential for development of epilepsy modifying therapies targeting intracellular pathways that broadly regulate gene expression & may be more efficacious than those that target the products of single gene candidates in isolation.

Public Health Relevance

Epilepsy affects over 3 million Americans and 65 million people worldwide. Over 30% of patients with epilepsy have seizures that cannot be controlled with current treatments and up to 50,000 people die each year from seizures and related causes. The proposed studies seek to find new ways of treating and preventing epilepsy by identifying, and then reversing, the complex molecular changes in brain cells that lead to this condition.

Agency
National Institute of Health (NIH)
Institute
National Institute of Neurological Disorders and Stroke (NINDS)
Type
Research Project (R01)
Project #
2R01NS051710-11A1
Application #
9052549
Study Section
Clinical Neuroplasticity and Neurotransmitters Study Section (CNNT)
Program Officer
Whittemore, Vicky R
Project Start
2005-04-01
Project End
2019-06-30
Budget Start
2015-09-01
Budget End
2016-06-30
Support Year
11
Fiscal Year
2015
Total Cost
Indirect Cost
Name
University of Colorado Denver
Department
Pediatrics
Type
Schools of Medicine
DUNS #
041096314
City
Aurora
State
CO
Country
United States
Zip Code
80045
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