CHK1 Inhibitor Blocks Phosphorylation of FAM122A and Promotes Replication Stress

Mol Cell. 2020 Nov 5;80(3):410-422.e6. doi: 10.1016/j.molcel.2020.10.008. Epub 2020 Oct 26.

Abstract

While effective anti-cancer drugs targeting the CHK1 kinase are advancing in the clinic, drug resistance is rapidly emerging. Here, we demonstrate that CRISPR-mediated knockout of the little-known gene FAM122A/PABIR1 confers cellular resistance to CHK1 inhibitors (CHK1is) and cross-resistance to ATR inhibitors. Knockout of FAM122A results in activation of PP2A-B55α, a phosphatase that dephosphorylates the WEE1 protein and rescues WEE1 from ubiquitin-mediated degradation. The resulting increase in WEE1 protein expression reduces replication stress, activates the G2/M checkpoint, and confers cellular resistance to CHK1is. Interestingly, in tumor cells with oncogene-driven replication stress, CHK1 can directly phosphorylate FAM122A, leading to activation of the PP2A-B55α phosphatase and increased WEE1 expression. A combination of a CHK1i plus a WEE1 inhibitor can overcome CHK1i resistance of these tumor cells, thereby enhancing anti-cancer activity. The FAM122A expression level in a tumor cell can serve as a useful biomarker for predicting CHK1i sensitivity or resistance.

Keywords: CHK1 inhibitor; CRISPR sgRNA screening; FAM122A; Fanconi Anemia; PABIR1; PP2A; WEE1.

Publication types

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

MeSH terms

  • Animals
  • Apoptosis / drug effects
  • Cell Cycle Checkpoints
  • Cell Cycle Proteins / metabolism
  • Cell Line
  • Cell Line, Tumor
  • Cell Proliferation / drug effects
  • Checkpoint Kinase 1 / antagonists & inhibitors
  • Checkpoint Kinase 1 / genetics*
  • Checkpoint Kinase 1 / metabolism
  • DNA Damage / drug effects
  • G2 Phase Cell Cycle Checkpoints / drug effects
  • Humans
  • Intracellular Signaling Peptides and Proteins / metabolism*
  • Intracellular Signaling Peptides and Proteins / physiology
  • Nuclear Proteins / metabolism
  • Phosphoproteins / metabolism*
  • Phosphoproteins / physiology
  • Phosphorylation
  • Protein Kinase Inhibitors / pharmacology
  • Protein Processing, Post-Translational / drug effects
  • Protein-Tyrosine Kinases / genetics
  • Pyrazines / metabolism
  • Pyrazines / pharmacology*
  • Pyrazoles / metabolism
  • Pyrazoles / pharmacology*
  • Signal Transduction / drug effects

Substances

  • Cell Cycle Proteins
  • Intracellular Signaling Peptides and Proteins
  • Nuclear Proteins
  • PABIR1 protein, human
  • Phosphoproteins
  • Protein Kinase Inhibitors
  • Pyrazines
  • Pyrazoles
  • prexasertib
  • Protein-Tyrosine Kinases
  • Checkpoint Kinase 1