Journal Article

Mathematical modeling suggests 14-3-3 proteins modulate RAF paradoxical activation

Tong, Melinda, Shokhirev, Maxim N., Mendiratta, Gaurav, Yu, Jingting, Li, Yao-Cheng, Wahl, Geoffrey M., Abbott, Kodye, Huang, Jianfeng, Carlip, Peter, Stites, Edward C., McFall, Thomas

2025

Overview

RAF inhibitor “paradoxical activation” (PA) is a phenomenon where RAF kinase inhibitors increase RAF kinase signaling. Through mathematical modeling and experimental data analysis, we recently demonstrated that the combination of conformational autoinhibition (CA) with the disruption of CA by RAF inhibitors plays an important role in PA. 14-3-3 proteins are known to modulate RAF CA and RAF dimerization. We here extend our mathematical model to include both roles of 14-3-3 proteins, and we derive rigorous analytical expressions of RAF signal regulation as modulated by 14-3-3 proteins. We then use the model to investigate how 14-3-3 proteins may modulate PA. We mathematically show 14-3-3 protein stabilization of the autoinhibited form of RAF should potentiate PA, while 14-3-3 protein stabilization of the active RAF dimer should reduce PA. Our analysis suggests that the net-effect will often be a potentiation of PA, and that 14-3-3 proteins may be capable of inducing PA for RAF inhibitors that normally show little to no PA. We test model-based insights experimentally with two different approaches: forced increases in 14-3-3 expression (which we find amplifies PA) and evolved resistance assays (which suggest increased 14-3-3 expression may contribute to resistance to RAF inhibitors). Overall, this work supports a role for 14-3-3 in modulating RAF-inhibitor mediated paradoxical activation.

• Publisher: Public Library of Science,PLOS,Public Library of Science (PLoS)
• Subject: 14-3-3 protein / 14-3-3 Proteins - chemistry / 14-3-3 Proteins - metabolism / Biology and Life Sciences / Cancer / Computational Biology / Data analysis / Dimerization / Enzyme Activation / Humans / Inhibitors / Kinases / Mathematical models / Medical research / Modelling / Models, Biological / Physical Sciences / Protein Kinase Inhibitors - pharmacology / Proteins / Proto-Oncogene Proteins c-raf - metabolism / raf Kinases - antagonists & inhibitors / raf Kinases - chemistry / raf Kinases - metabolism / Raf protein / Research and Analysis Methods / Roles / Signal Transduction / Stabilization