Mechanistic MALDI-TOF Cell-Based Assay for the Discovery of Potent and Specific Fatty Acid Synthase Inhibitors.

Abstract

Human cancers require fatty acid synthase (FASN)-dependent de novo long-chain fatty acid synthesis for proliferation. FASN is therefore an attractive drug target, but fast technologies for reliable label-free cellular compound profiling are lacking. Recently, MALDI-mass spectrometry (MALDI-MS) has emerged as an effective technology for discovery of recombinant protein target inhibitors. Here we present an automated, mechanistic MALDI-MS cell assay, which monitors accumulation of the FASN substrate, malonyl-coenzyme A (CoA), in whole cells with limited sample preparation. Profiling of inhibitors, including unpublished compounds, identified compound 1 as the most potent FASN inhibitor (1 nM in A549 cells) discovered to date. Moreover, cellular MALDI-MS assays enable parallel profiling of additional pathway metabolites. Surprisingly, several compounds triggered cytidine 5'-diphosphocholine (CDP-choline) but not malonyl-CoA accumulation indicating that they inhibit diacylglycerol generation but not FASN activity. Taken together, our study suggests that MALDI-MS cell assays may become important tools in drug profiling that provide additional mechanistic insights concerning compound action on metabolic pathways.

• Authors: David Weigt, Cynthia A Parrish, Julie A Krueger, Catherine A Oleykowski, Alan R Rendina, Carsten Hopf
• Journal: Cell chemical biology (Cell Chem Biol) Vol. 26 Issue 9 Pg. 1322-1331.e4 (09 19 2019) ISSN: 2451-9448 [Electronic] United States
• PMID: 31279605 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
• Copyright: Copyright © 2019 Elsevier Ltd. All rights reserved.

Chemical References

• Malonyl Coenzyme A
• FASN protein, human
• Fatty Acid Synthase, Type I
• Fatty Acid Synthases

Topics

• A549 Cells
• Apoptosis (drug effects)
• Cell Line, Tumor
• Drug Discovery (methods)
• Drug Evaluation, Preclinical (methods)
• Fatty Acid Synthase, Type I (antagonists & inhibitors, metabolism)
• Fatty Acid Synthases (antagonists & inhibitors, metabolism)
• Humans
• Inhibitory Concentration 50
• K562 Cells
• Lipogenesis
• Malonyl Coenzyme A (metabolism)
• Proof of Concept Study
• Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization (methods)