400 Potential Drug Therapy for Fragile X Tremor/Ataxia Syndrome

Abstract

OBJECTIVES/GOALS: Fragile X-associated tremor/ataxia syndrome (FXTAS) is a devastating rare neurological disorder that negatively impacts movement and cognition. To date, there are no effective pharmacological treatments for FXTAS. Our goal was to develop a cell culture model of FXTAS to investigate promising therapeutics. METHODS/STUDY POPULATION: To establish mitochondrial dysfunction, normal human cell lines and human-induced pluripotent cells were treated with multiple concentrations of glucose/ glucose oxidase (GluOx) at 2,12, and 24 hour time points to induce varying intensities of oxidative stress. The degrees of oxidative stress were measured by apoptosis and mitochondrial reactive oxygen species (ROS) production. Curcumin and MSKE compounds effective against oxidative damage in mitochondria were used to rescue glucose oxidase-induced oxidative damage in both cell lines. To test the ability of these drugs to restore mitochondrial health, cell viability and cellular superoxide production were assessed by propidium iodide and the MitoSox fluorescence assay, respectively. RESULTS/ANTICIPATED RESULTS: We anticipated that GluOx at varying concentrations and time points would proportionally increase levels of apoptosis and mitochondrial ROS, reflective of mitochondrial damage, with the most severe dysfunction occurring at a dose of 25 nM and the longest duration of 24-hr exposure. Administration of MSKE in concentrations ranging from 10-8 to 10-5 M in half log increments, did not reverse the oxidative defects induced in the cell lines. However, curcumin concentrations increased cell viability at the 2, 12, and 24 hour time period. Results indicate that the research design should be modified by increasing the concentration of both glucose and MSKE to provide a reliable test of the hypothesis. DISCUSSION/SIGNIFICANCE: These studies illustrate the usefulness of this in vitro model to test novel therapeutics in neuronal FXTAS models and expand the discovery of mitochondrial markers for the syndrome.