Phenotype-Based Screens with Conformation-Specific Inhibitors Reveal p38 Gamma and Delta as Targets for HCC Polypharmacology

Mol Cancer Ther. 2019 Sep;18(9):1506-1519. doi: 10.1158/1535-7163.MCT-18-0571. Epub 2019 Jun 18.

Abstract

The approved kinase inhibitors for hepatocellular carcinoma (HCC) are not matched to specific mutations within tumors. This has presented a daunting challenge; without a clear target or mechanism, no straightforward path has existed to guide the development of improved therapies for HCC. Here, we combine phenotypic screens with a class of conformation-specific kinase inhibitors termed type II to identify a multikinase inhibitor, AD80, with antitumoral activity across a variety of HCC preclinical models, including mouse xenografts. Mass spectrometry profiling found a number of kinases as putative targets for AD80, including several receptor and cytoplasmic protein kinases. Among these, we found p38 gamma and delta as direct targets of AD80. Notably, a closely related analog of AD80 lacking p38δ/γ activity, but retaining several other off-target kinases, lost significant activity in several HCC models. Moreover, forced and sustained MKK6 → p38→ATF2 signaling led to a significant reduction of AD80 activity within HCC cell lines. Together with HCC survival data in The Cancer Genome Atlas and RNA-seq analysis, we suggest p38 delta and gamma as therapeutic targets in HCC and an "AD80 inhibition signature" as identifying those patients with best clinical outcomes.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Carcinoma, Hepatocellular / drug therapy*
  • Carcinoma, Hepatocellular / pathology
  • Cell Line, Tumor
  • Drug Screening Assays, Antitumor / methods
  • Female
  • Heterocyclic Compounds, 4 or More Rings / chemistry
  • Heterocyclic Compounds, 4 or More Rings / pharmacokinetics
  • Heterocyclic Compounds, 4 or More Rings / pharmacology*
  • Humans
  • Kaplan-Meier Estimate
  • Liver Neoplasms / drug therapy*
  • Liver Neoplasms / pathology
  • Male
  • Mice, Inbred C57BL
  • Mice, Nude
  • Mitogen-Activated Protein Kinase 12 / antagonists & inhibitors*
  • Mitogen-Activated Protein Kinase 12 / chemistry
  • Mitogen-Activated Protein Kinase 13 / antagonists & inhibitors*
  • Mitogen-Activated Protein Kinase 13 / chemistry
  • Phenotype
  • Polypharmacology
  • Xenograft Model Antitumor Assays / methods*

Substances

  • AD80 compound
  • Heterocyclic Compounds, 4 or More Rings
  • Mitogen-Activated Protein Kinase 12
  • Mitogen-Activated Protein Kinase 13