Developing a High-Throughput Drug Screening Method to Find DEN1 Inhibitors Using a Luminescent Biosensor — 96p — Isabella Veffredo, Yun-Seok Choi

Nedd8, a post-translational modifier, regulates cellular signals through protein conjugation. Deneddylases recycle conjugated Nedd8 back to its free form. Imbalances between free and conjugated Nedd8, caused by enzyme defects or cellular stress, are implicated in various diseases. DEN1, a deneddylase, has emerged as a potential drug target for treating amyotrophic lateral sclerosis (ALS) and overcoming chemotherapeutic resistance in acute lymphoblastic leukemia (ALL). Developing an efficient method to monitor DEN1 activity is crucial for discovering DEN1 inhibitors. This study aims to establish a high-throughput screening method for DEN1 inhibitors using eDEN1-NanoBiT, a luminescent sensor that selectively detects free Nedd8, the product of DEN1 activity. To optimize the large-scale screening process, we focused on minimizing the concentration of each ingredient in the DEN1 reaction. Furthermore, we confirmed the compatibility of the screening reaction with 6% DMSO conditions, enhancing its practicality for drug discovery efforts. The impact of this research lies in its potential to significantly advance drug discovery for ALS and ALL.

Black Hills State University
Yun-Seok Choi