?Stroke is a leading cause of morbidity and mortality in the United States. However less than 20% of patients are eligible for the current approved interventions of tissue plasminogen activator or thrombectomy. We seek to develop new therapeutics to reduce the extent of damage and functional impairment resulting from ischemic injury to the brain, an area of significant unmet medical need. We have found that interferon regulatory factor (IRF) mediated gene transcription may represent an endogenous mechanism of neuroprotection that is associated with a reduction in ischemic injury. Using both cell and mouse models of stroke we have demonstrated that administration of compounds following the ischemic insult, that are known to induce IRF mediated gene transcription significantly reduces the extent of damage. These results indicate that activation of IRF transcription factors following stroke may be a viable therapeutic intervention for the treatment of stroke patients. The ultimate goal of this STTR program is to identify IRF activators with minimal off-target immune activity and specificity for both mouse and human for preclinical development for stroke. Our specific goal for this Phase I application is to develop and validate a high-throughput screening platform to identify clinically viable IRF activating compounds for further development. We propose the following aims: Aim 1: Validate a high-throughput primary screening assay to identify potent and selective IRF activators using a human THP1 dual ISRE/NF?B reporter cell line and 384-well format. Aim 2: Validate a high-throughput secondary screening assay in mouse J774 cells containing dual ISRE/NFkB reporters to evaluate cross-species activity of primary hits from Aim 1. Aim 3: Verify the target and species selectivity of secondary hits by evaluating a panel of downstream IRF-inducible genes in human and mouse brain-derived cells.
Public Health Relevance Statement: PUBLIC HEALTH RELEVANCE Stroke is a leading cause of morbidity and mortality in the United States with approximately 800,000 cases a year reported. This proposal seeks to establish a high throughput screen for identification of compounds that activate interferon regulatory factors as potential mediators of neuroprotection, with the ultimate goal being the development of a stroke therapeutic.
NIH Spending Category: Biotechnology; Brain Disorders; Neurosciences; Stroke
Project Terms: Alteplase; analog; Animals; Area; base; Biological Assay; Brain; brain cell; Brain Injuries; Cell Death; Cell Line; Cell model; Cells; Cessation of life; Clinical; cytotoxic; cytotoxicity; Data; deprivation; Development; Dimethylxanthenone Acetic Acid; Dose; effective therapy; Event; functional disability; gene induction; Genes; Genetic Transcription; Glucose; Goals; high throughput screening; Human; Immune Targeting; in vitro activity; Injury; Interferon Activation; interferon regulatory factor 10; Interferon Regulatory Factor 2; Interferons; Intervention; IRF3 gene; Ischemia; Ischemic Brain Injury; Lead; Libraries; Mediating; Mediator of activation protein; Medical; Molecular; Morbidity - disease rate; Mortality Vital Statistics; mouse model; Mus; neuroprotection; novel; novel therapeutics; Oxygen; Pathway interactions; Patients; Pattern; Pharmaceutical Chemistry; Pharmacological Treatment; Phase; phase 2 study; pre-clinical; preconditioning; Process; programs; Proteins; public health relevance; Reagent; Reporter; Reporting; Reproducibility; Research; Research Design; response; screening; Signal Transduction; Small Business Technology Transfer Research; small molecule; Source; Specificity; Stress; stroke; Structure-Activity Relationship; Testing; Therapeutic; Therapeutic Intervention; Thrombectomy; Time; Titrations; transcription factor; Transcription factor genes; Tumor Necrosis Factor-alpha; Undifferentiated; United States; Validation
