Brain swelling is a serious complication of multiple disease conditions including liver failure, metastatic tumors, traumatic brain injury and ischemic stroke. TBI and stroke afflict 1.4M and 700K persons per year in the US alone, and is a costly health care burden and a devastating social burden. Current treatments for brain swelling are limited and generally ineffective, highlighting the dramatic unmet need for better therapeutics. A better understanding of the molecular pathways and cellular mechanisms is sorely needed to identify new drug targets as well as more predictive biomarkers that can stratify patients for clinical treatment decisions. The goal of this project is to identify new drug targets and biomarkers of response for cytotoxic edema of astrocytes. We propose to use a novel and innovative technology that we have developed that will take an unbiased approach to functionally identifying the causal mediators of astrocyte swelling. Our approach uses a large panel of genetically diverse astrocyte lines to identify the genes and pathways that mechanistically underlie cytotoxic edema. In Phase I, we will develop two high throughput kinetic assays for astrocyte swelling that are robust, scalable and automatable for screening compounds that induce or block swelling. An impedance based morphological assay will measure swelling and recovery, and a rapid calcium flux assay will measure cationic influx. In Phase II we will use these assays to screen ~300 genetically diverse astrocyte lines, and then map and identify the genes that mediate response to inducers and blockers of cell swelling. Validation of candidate target genes will be conducted in both human and mouse astrocytes. Human genes that modify human astrocyte response to compound are the ultimate aims and end-products of this project. Those genes and sequence variants in the human population will also be evaluated as potential prognostic biomarkers using existing clinical data in retrospective analyses.
Public Health Relevance Statement: Brain swelling (cerebral edema) is a serious consequence of many disorders such as acute liver failure, metastatic cancer, stroke and traumatic brain injury. A more complete understanding of the molecular pathways underlying cerebral edema is a critical bottleneck in the development of new therapies for brain swelling. We want to understand the molecular mechanisms underlying swelling in astrocytes, the most abundant cell type in the brain, toward the discovery of more effective treatments.
Project Terms: abstracting; Acute Liver Failure; Adrenal Cortex Hormones; Astrocytes; base; Biological Assay; Biology; Biotechnology; Blood - brain barrier anatomy; Blood Vessels; Brain; brain cell; Brain Edema; brain volume; candidate validation; care burden; Cations; cell type; Cell Volumes; Cells; Cephalic; Cerebral Edema; Clinical; Clinical Data; Clinical Treatment; Collaborations; commercialization; Complication; cytotoxic; Data; Development; Disease; Disseminated Malignant Neoplasm; drug development; drug discovery; Drug Targeting; Edema; effective therapy; electric impedance; Endothelial Cells; Etiology; Evaluation; Extracellular Space; Extravasation; Gene Targeting; Gene-Modified; Genes; Genetic; Genetic Screening; genome-wide; genomic tools; Goals; Government; Head; Healthcare; Heritability; Human; In Vitro; innovation; innovative technologies; Intervention; Intracranial Hypertension; Ischemic Stroke; Kinetics; Lead; Liver Failure; Maps; Measures; Mediating; Mediator of activation protein; Molecular; Mus; Neoplasm Metastasis; new technology; new therapeutic target; novel; novel therapeutics; Outcome; Pathway interactions; patient stratification; Perfusion; Persons; Pharmaceutical Preparations; Phase; Phenotype; Population; Potassium; pre-clinical; predictive marker; Prognostic Marker; QTL Genes; Recovery; release of sequestered calcium ion into cytoplasm; response; screening; social; Sodium; stroke; Swelling; targeted biomarker; Testing; Therapeutic; Tight Junctions; transcriptomics; Traumatic Brain Injury; Variant; Variation (Genetics); Water
