Drug Discovery for AMN

Sponsored by the Korengold AMN Research Fund.

The overall goal of this research project is to accelerate the pace at which new therapies for the most common inherited peroxisomal disorder, adrenomyeloneuropathy (AMN), are discovered and tested in clinical trials through the application of existing cutting-edge drug-screening and validation technologies. Due to a defect in the ALD protein (ALDP) encoded by the ABCD1 gene, cultured skin fibroblasts from AMN patients accumulate very long chain fatty acids that can be measured by liquid chromatography tandem mass spectrometry (LC-MS/MS). This study will harness LC-MS/MS to rapidly screen chemical libraries for the ability to rescue ALDP function in AMN patient fibroblasts using the cellular levels of the complex lipid C26:0-lysophosphorylcholine (C26:0-LPC) as a biomarker for ALDP function. Afterwards, researchers will test the most promising compounds for their ability to rescue peroxisome activity in cellular and mouse models of these disorders. In sum, this will provide a rapid and highly-efficient means of identifying promising candidate drug treatments for AMN patients.

First, researchers will optimize the conditions needed for the seeding of patient fibroblasts in microtiter plates, addition of test compounds, and extraction of lipids prior to automatic injection on the LC-MS/MS (Year 1). Once this process is established and rigorous quality control standards are met, we will screen a 2,000 compound library of bioactive compounds at two concentrations using an ABCD1-mutant AMN model cell line (Year 1). After completing this optimization phase, the same ABCD1-mutant cell line will be used to screen a library of 10,000 compounds likely to pass the blood-brain barrier (Year 2). Positive compounds identified by our screens will be re-tested and subjected to additional confirmation (Years 2-3). Finally, we will test the efficacy of the most promising drugs to rescue peroxisome function in an existing ABCD1-knockout AMN mouse model (Year 3). Overall, researchers will develop a flexible and highly-efficient screening strategy to identify compounds that would enter a pipeline for drug development and ultimately, treatment of patients with AMN.