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Exploring the synergy between PARP and CHK1 inhibition in matched BRC2 mutant and corrected cells

Lookup NU author(s): Hannah Smith, Dr Lisa PrendergastORCiD, Professor Nicola CurtinORCiD



This work is licensed under a Creative Commons Attribution 4.0 International License (CC BY 4.0).


PARP inhibition results in the accumulation of DNA SSBs, causing replication stress (RS) and lesions that can only be resolved by homologous recombination repair (HRR). Defects in HRR, e.g., due to BRCA2 mutation, confer profound sensitivity to PARP inhibitor (PARPi) cytotoxicity. In response to RS, CHK1 is activated to signal to S and G2/M cell cycle checkpoints and also to HRR. To determine the relative contribution of these two functions of CHK1 to survival following PARPi exposure, we investigated the effects of rucaparib (a PARPi) and PF-477736 (a CHK1 inhibitor) alone and in combination in cells with mutated and corrected BRCA2. The BRCA2 mutated V-C8 cells were 1000× more sensitive to rucaparib cytotoxicity than their matched BRCA2 corrected V-C8.B2 cells, but no more sensitive to PF-477736 despite having seven-fold higher levels of RS. PF-477736 caused a five-fold enhancement of rucaparib cytotoxicity in the V-C8.B2 cells, but no enhancement in the V-C8 cells. This differential sensitivity was not due to a difference in PARP1 or CHK1 expression or activity. PF-477736 increased rucaparib-induced RS (γH2AX foci) and completely inhibited RAD51 focus formation, indicating a profound suppression of HRR. Our data suggested that inhibition of HRR was the main mechanism of sensitisation to rucaparib, compounded with an inhibition of cell cycle checkpoints by PF-477736.

Publication metadata

Author(s): Smith H, Prendergast L, Curtin NJ

Publication type: Article

Publication status: Published

Journal: Cancers

Year: 2020

Volume: 12

Issue: 4

Online publication date: 04/04/2020

Acceptance date: 31/03/2020

Date deposited: 15/04/2020

ISSN (print): 2072-6694

ISSN (electronic): 2072-6694

Publisher: MDPI


DOI: 10.3390/cancers12040878


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