Genome-wide and high-density CRISPR-Cas9 screens identify point mutations in<i>PARP1</i>causing PARP inhibitor resistance

Pettitt, Stephen J.; Krastev, Dragomir B.; Brandsma, Inger; Dréan, Amy; Song, Feifei; Aleksandrov, Radoslav; Menon, Malini; Brough, Rachel; Campbell, James; Frankum, Jessica; Ranes, Michael; Pemberton, Helen; Rafiq, Rumana; Fenwick, Kerry; Swain, Amanda; Guettler, Sebastian; Lee, Jungmin; Swisher, Elizabeth M.; Stoynov, Stoyno; Yusa, Kosuke; Ashworth, Alan; Lord, Christopher J.

doi:10.1101/203224

Cited by 29 publications

(43 citation statements)

References 51 publications

(88 reference statements)

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“…The RAD51 assay has some limitations: firstly, when PARPi sensitivity occurs via mechanisms that do not directly impact on the ability of cells to perform HRR, e.g., alterations in ATM (Chen et al , ; Davies et al , ; preprint: Balmus et al , ) or in the RNASEH2 complex (Zimmermann et al , ); secondly, when PARPi sensitivity occurs via mechanisms that preserve RAD51 foci formation, e.g., alterations in the MRN complex, RAD51AP1, polymerase eta, or ERCC1 (Kawamoto et al , ; Wiese et al , ; Oplustilova et al , ; Postel‐Vinay et al , ); thirdly, when HRR‐deficient tumors have acquired PARPi resistance via RAD51‐independent mechanisms such as loss of PARG, mutations in PARP1, or those that involve replication fork stabilization (Guillemette et al , ; Chaudhuri et al , ; Kais et al , ; Yazinski et al , ; Gogola et al , ; Michelena et al , ; Pettitt et al , ); fourthly, when a tumor has low proliferation index or low endogenous DNA damage, in which cases the assay would not be feasible.…”

Section: Discussionmentioning

confidence: 99%

“…or in the RNASEH2 complex(Zimmermann et al, 2018); secondly, when PARPi sensitivity occurs via mechanisms that preserve RAD51 foci formation, e.g., alterations in the MRN complex, RAD51AP1, polymerase eta, or ERCC1(Kawamoto et al, 2005;Wiese et al, 2007;Oplustilova et al, 2012;Postel-Vinay et al, 2013); thirdly, when HRR-deficient tumors have acquired PARPi resistance via RAD51independent mechanisms such as loss of PARG, mutations in PARP1, or those that involve replication fork stabilization(Guillemette et al, 2015;Chaudhuri et al, 2016;Kais et al, 2016; Yazinski et al, 2017;Gogola et al, 2018;Michelena et al, 2018;Pettitt et al, 2018);…”

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confidence: 99%

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A RAD 51 assay feasible in routine tumor samples calls PARP inhibitor response beyond BRCA mutation

Castroviejo-Bermejo¹,

et al. 2018

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Poly(ADP‐ribose) polymerase (PARP) inhibitors (PARPi) are effective in cancers with defective homologous recombination DNA repair (HRR), including BRCA1/2‐related cancers. A test to identify additional HRR‐deficient tumors will help to extend their use in new indications. We evaluated the activity of the PARPi olaparib in patient‐derived tumor xenografts (PDXs) from breast cancer (BC) patients and investigated mechanisms of sensitivity through exome sequencing, BRCA1 promoter methylation analysis, and immunostaining of HRR proteins, including RAD51 nuclear foci. In an independent BC PDX panel, the predictive capacity of the RAD51 score and the homologous recombination deficiency (HRD) score were compared. To examine the clinical feasibility of the RAD51 assay, we scored archival breast tumor samples, including PALB2‐related hereditary cancers. The RAD51 score was highly discriminative of PARPi sensitivity versus PARPi resistance in BC PDXs and outperformed the genomic test. In clinical samples, all PALB2‐related tumors were classified as HRR‐deficient by the RAD51 score. The functional biomarker RAD51 enables the identification of PARPi‐sensitive BC and broadens the population who may benefit from this therapy beyond BRCA1/2‐related cancers.

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Section: Discussionmentioning

confidence: 99%

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confidence: 99%

A RAD 51 assay feasible in routine tumor samples calls PARP inhibitor response beyond BRCA mutation

Castroviejo-Bermejo¹,

et al. 2018

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“…Then, we silenced the PARP1 gene in RD-ES and SK-ES-1 cells and proved the dependency of PARPis' cytotoxicity on PARP1 in both cell lines. 42 For example, PARP1 mutations are present in talazoparibresistant BRCA1-deficient SUM149 and COV362 cells, and a R591C mutation was associated with olaparib resistance in an ovarian cancer patient. Complementation of the KO cells with WT-or E988K-PARP1 showed that the cytotoxicity and PARP1-DNA trapping induced by talazoparib were dependent on the polymerase activity of PARP1, because purified E988K-PARP1 and WT-PARP1 proteins had similar capabilities to bind NAD + , PARPi and DNA.…”

Section: Discussionmentioning

confidence: 99%

“…On the other hand, the mutations reducing PARP1 polymerase activity have been found to cause cellular and clinical resistance to PARPis. 42 For example, PARP1 mutations are present in talazoparibresistant BRCA1-deficient SUM149 and COV362 cells, and a R591C mutation was associated with olaparib resistance in an ovarian cancer patient.…”

Section: Discussionmentioning

confidence: 99%

Increased PARP1‐DNA binding due to autoPARylation inhibition of PARP1 on DNA rather than PARP1‐DNA trapping is correlated with PARP1 inhibitor's cytotoxicity

Chen

Wang

et al. 2019

Intl Journal of Cancer

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PARP1 inhibitors (PARPis) are used clinically during cancer therapy and are thought to exert their cytotoxicity through PARP1 polymerase inhibition and PARP1‐DNA trapping. Here, we showed no significant correlation between PARP1‐DNA trapping and cytotoxicity induced by PARPis. We complemented PARP1‐knockout sublines with wild‐type PARP1 and 11 mutants with different point mutations that affect the polymerase activity. When examining the PARPi talazoparib, the induced cytotoxicity was highly significantly correlated with cellular PARP1 polymerase activity, but not with its PARP1‐DNA trapping or polymerase inhibition. Similarly, talazoparib's PARP1‐DNA trapping revealed significant correlation with the polymerase activity rather than its inhibition. Differently, however, when evaluating purified wild‐type and mutated PARP1, we identified an almost linear relationship between PARPis’ inhibiting PARP1 dissociation from DNA and their cytotoxicity in 17 cancer cell lines. In contrast, no significant correlation existed between PARP1 polymerase inhibition in the histone‐based systems and the cytotoxicity. After careful comparisons on different methods and detection targets, we conclude that the PARPi‐mediated increase in PARP1‐DNA binding by inhibiting autoPARylation of PARP1 on DNA rather than in PARP1‐DNA trapping is correlated with PARPi's cytotoxicity. Accordingly, we established a new PARPi screening model that more closely predicts cytotoxicity.

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“…Furthermore, cells with PARP1 mutations were 100‐fold more resistant to PARP inhibitors than were cells with wild‐type PARP1 . Mutations both within and outside the PARP1 DNA‐binding domains alter PARP1 trapping and induce PARP inhibitor resistance . Cancer cells may up‐regulate the HR repair pathway to compensate for the loss of BER as a result of PARP1 inhibition, and the HR and BER pathways interact to regulate cancer cell viability via decreased PARP1 and increased RAD51 expression levels.…”

Section: Mechanisms Underlying Parp Inhibitor Resistancementioning

confidence: 99%

PARP inhibitors in ovarian cancer: Sensitivity prediction and resistance mechanisms

Jiang

et al. 2019

J Cellular Molecular Medi

131

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Poly (ADP‐ribose) polymerase (PARP) inhibitors have provided great clinical benefits to ovarian cancer patients. To date, three PARP inhibitors, namely, olaparib, rucaparib and niraparib have been approved for the treatment of ovarian cancer in the United States. Homologous recombination deficiency (HRD) and platinum sensitivity are prospective biomarkers for predicting the response to PARP inhibitors in ovarian cancers. Preclinical data have focused on identifying the gene aberrations that might generate HRD and induce sensitivity to PARP inhibitors in vitro in cancer cell lines or in vivo in patient‐derived xenografts. Clinical trials have focused on genomic scar analysis to identify biomarkers for predicting the response to PARP inhibitors. Additionally, researchers have aimed to investigate mechanisms of resistance to PARP inhibitors and strategies to overcome this resistance. Combining PARP inhibitors with HR pathway inhibitors to extend the utility of PARP inhibitors to BRCA‐proficient tumours is increasingly foreseeable. Identifying the population of patients with the greatest potential benefit from PARP inhibitor therapy and the circumstances under which patients are no longer suited for PARP inhibitor therapy are important. Further studies are required in order to propose better strategies for overcoming resistance to PARP inhibitor therapy in ovarian cancers.

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Genome-wide and high-density CRISPR-Cas9 screens identify point mutations inPARP1causing PARP inhibitor resistance

Cited by 29 publications

References 51 publications

A RAD 51 assay feasible in routine tumor samples calls PARP inhibitor response beyond BRCA mutation

A RAD 51 assay feasible in routine tumor samples calls PARP inhibitor response beyond BRCA mutation

Increased PARP1‐DNA binding due to autoPARylation inhibition of PARP1 on DNA rather than PARP1‐DNA trapping is correlated with PARP1 inhibitor's cytotoxicity

PARP inhibitors in ovarian cancer: Sensitivity prediction and resistance mechanisms

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