Discovering Novel Drug Targets for CNS Edema by In Vitro Genetics

Abstract 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 M and K 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 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 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