Inactive Parp2 causes Tp53-dependent lethal anemia by blocking replication-associated nick ligation in erythroblasts

Mol Cell. 2024 Oct 17;84(20):3916-3931.e7. doi: 10.1016/j.molcel.2024.09.020. Epub 2024 Oct 8.

Abstract

Poly (ADP-ribose) polymerase (PARP) 1 and 2 enzymatic inhibitors (PARPi) are promising cancer treatments. But recently, their use has been hindered by unexplained severe anemia and treatment-related leukemia. In addition to enzymatic inhibition, PARPi also trap PARP1 and 2 at DNA lesions. Here we report that, unlike Parp2-/- mice, which develop normally, mice expressing catalytically inactive Parp2 (E534A and Parp2EA/EA) succumb to Tp53- and Chk2-dependent erythropoietic failure in utero, mirroring Lig1-/- mice. While DNA damage mainly activates PARP1, we demonstrate that DNA replication activates PARP2 robustly. PARP2 is selectively recruited and activated by 5'-phosphorylated nicks (5'p-nicks), including those between Okazaki fragments, resolved by ligase 1 (Lig1) and Lig3. Inactive PARP2, but not its active form or absence, impedes Lig1- and Lig3-mediated ligation, causing dose-dependent replication fork collapse, which is detrimental to erythroblasts with ultra-fast forks. This PARylation-dependent structural function of PARP2 at 5'p-nicks explains the detrimental effects of PARP2 inactivation on erythropoiesis, shedding light on PARPi-induced anemia and the selection for TP53/CHK2 loss.

Keywords: 5′p-nicks; Lig1; PARP inhibition; PARP inhibitor; PARP2; anemia; erythropoiesis; hematological toxicity.

MeSH terms

  • Anemia* / chemically induced
  • Anemia* / genetics
  • Anemia* / pathology
  • Animals
  • Checkpoint Kinase 2* / genetics
  • Checkpoint Kinase 2* / metabolism
  • DNA Damage
  • DNA Ligase ATP / genetics
  • DNA Ligase ATP / metabolism
  • DNA Replication* / drug effects
  • Erythroblasts* / drug effects
  • Erythroblasts* / metabolism
  • Erythropoiesis / drug effects
  • Erythropoiesis / genetics
  • Female
  • Humans
  • Mice
  • Mice, Inbred C57BL
  • Mice, Knockout
  • Poly (ADP-Ribose) Polymerase-1 / antagonists & inhibitors
  • Poly (ADP-Ribose) Polymerase-1 / genetics
  • Poly (ADP-Ribose) Polymerase-1 / metabolism
  • Poly(ADP-ribose) Polymerase Inhibitors / pharmacology
  • Poly(ADP-ribose) Polymerases* / genetics
  • Poly(ADP-ribose) Polymerases* / metabolism
  • Tumor Suppressor Protein p53* / genetics
  • Tumor Suppressor Protein p53* / metabolism

Substances

  • Tumor Suppressor Protein p53
  • Poly(ADP-ribose) Polymerases
  • Parp2 protein, mouse
  • Checkpoint Kinase 2
  • Trp53 protein, mouse
  • DNA Ligase ATP
  • Chek2 protein, mouse
  • Poly (ADP-Ribose) Polymerase-1
  • Parp1 protein, mouse
  • Poly(ADP-ribose) Polymerase Inhibitors