Genome-wide and high-density CRISPR-Cas9 screens identify point mutations in PARP1 causing PARP inhibitor resistance.
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Date
2018-05-01ICR Author
Author
Pettitt, SJ
Krastev, DB
Brandsma, I
Dréan, A
Song, F
Aleksandrov, R
Harrell, MI
Menon, M
Brough, R
Campbell, J
Frankum, J
Ranes, M
Pemberton, HN
Rafiq, R
Fenwick, K
Swain, A
Guettler, S
Lee, J-M
Swisher, EM
Stoynov, S
Yusa, K
Ashworth, A
Lord, CJ
Type
Journal Article
Metadata
Show full item recordAbstract
Although PARP inhibitors (PARPi) target homologous recombination defective tumours, drug resistance frequently emerges, often via poorly understood mechanisms. Here, using genome-wide and high-density CRISPR-Cas9 "tag-mutate-enrich" mutagenesis screens, we identify close to full-length mutant forms of PARP1 that cause in vitro and in vivo PARPi resistance. Mutations both within and outside of the PARP1 DNA-binding zinc-finger domains cause PARPi resistance and alter PARP1 trapping, as does a PARP1 mutation found in a clinical case of PARPi resistance. This reinforces the importance of trapped PARP1 as a cytotoxic DNA lesion and suggests that PARP1 intramolecular interactions might influence PARPi-mediated cytotoxicity. PARP1 mutations are also tolerated in cells with a pathogenic BRCA1 mutation where they result in distinct sensitivities to chemotherapeutic drugs compared to other mechanisms of PARPi resistance (BRCA1 reversion, 53BP1, REV7 (MAD2L2) mutation), suggesting that the underlying mechanism of PARPi resistance that emerges could influence the success of subsequent therapies.
Subject
Cell Line, Tumor
Animals
Mice, Inbred BALB C
Humans
Mice
Mice, Nude
Neoplasms
Phthalazines
BRCA1 Protein
Xenograft Model Antitumor Assays
DNA Mutational Analysis
Mutagenesis
Zinc Fingers
Drug Resistance, Neoplasm
Point Mutation
Aged
Female
CRISPR-Cas Systems
Poly(ADP-ribose) Polymerase Inhibitors
Precision Medicine
Mouse Embryonic Stem Cells
Poly (ADP-Ribose) Polymerase-1
Whole Genome Sequencing
Research team
Development & Cancer
Gene Function
Structural Biology of Cell Signalling
Language
eng
Date accepted
2018-03-22
License start date
2018-05-10
Citation
Nature communications, 2018, 9 (1), pp. 1849 - ?
Publisher
NATURE PUBLISHING GROUP