Journal Article

Mutations in UBA3 Confer Resistance to the NEDD8-Activating Enzyme Inhibitor MLN4924 in Human Leukemic Cells

Maclean, Neil, Petroski, Matthew D., da Silva, Sara R., Paiva, Stacey-Lynn, Lukkarila, Julie L., Goard, Carolyn A., Xu, G. Wei, Hurren, Rose, Bhattacharjee, Rabindra N., Toth, Julia I., Sukhai, Mahadeo A., Gunning, Patrick T., Schimmer, Aaron D., Dhe-Paganon, Sirano

2014

Overview

The NEDD8-activating enzyme (NAE) initiates neddylation, the cascade of post-translational NEDD8 conjugation onto target proteins. MLN4924, a selective NAE inhibitor, has displayed preclinical anti-tumor activity in vitro and in vivo, and promising clinical activity has been reported in patients with refractory hematologic malignancies. Here, we sought to understand the mechanisms of resistance to MLN4924. K562 and U937 leukemia cells were exposed over a 6 month period to MLN4924 and populations of resistant cells (R-K562(MLN), R-U937(MLN)) were selected. R-K562(MLN) and R-U937(MLN) cells contain I310N and Y352H mutations in the NAE catalytic subunit UBA3, respectively. Biochemical analyses indicate that these mutations increase the enzyme's affinity for ATP while decreasing its affinity for NEDD8. These mutations effectively contribute to decreased MLN4924 potency in vitro while providing for sufficient NAE function for leukemia cell survival. Finally, R-K562(MLN) cells showed cross-resistance to other NAE-selective inhibitors, but remained sensitive to a pan-E1 (activating enzyme) inhibitor. Thus, our work provides insight into mechanisms of MLN4924 resistance to facilitate the development of more effective second-generation NAE inhibitors.

• Publisher: Public Library of Science,Public Library of Science (PLoS)
• Subject: Adenosine Triphosphate - chemistry / Adenosine Triphosphate - metabolism / Affinity / Analysis / Anticancer properties / Antineoplastic Agents - chemistry / Antineoplastic Agents - pharmacology / Antitumor agents / Apoptosis / Biology and Life Sciences / Cancer / Care and treatment / Catalysis / Cell Line, Tumor / Cell survival / Conjugation / Cross-resistance / Cullin Proteins - metabolism / Cyclopentanes - chemistry / Cyclopentanes - pharmacology / Deoxyribonucleic acid / Diagnosis / DNA / DNA Mutational Analysis / Drug Resistance, Neoplasm - genetics / Enzyme inhibitors / Enzyme Inhibitors - chemistry / Enzyme Inhibitors - pharmacology / Enzymes / Genetic aspects / Genotype / Health care networks / Humans / Inhibitors / K562 Cells / Kinases / Leukemia / Leukemia - genetics / Leukemia - metabolism / Lymphoma / Medical research / Medicine and Health Sciences / Models, Molecular / Mutation / NEDD8 Protein / Point Mutation / Post-translation / Protein Binding / Protein Conformation / Proteins / Pyrimidines - chemistry / Pyrimidines - pharmacology / Structure-Activity Relationship / U937 Cells / Ubiquitin / Ubiquitin-Activating Enzymes - antagonists & inhibitors / Ubiquitin-Activating Enzymes - chemistry / Ubiquitin-Activating Enzymes - genetics / Ubiquitins - genetics / Ubiquitins - metabolism