TY - JOUR
T1 - Inhibition of the ERCC1-XPF structure-specific endonuclease to overcome cancer chemoresistance
AU - McNeil, Ewan M.
AU - Astell, Katy R.
AU - Ritchie, Ann Marie
AU - Shave, Steven
AU - Houston, Douglas R.
AU - Bakrania, Preeti
AU - Jones, Hayley M.
AU - Khurana, Puneet
AU - Wallace, Claire
AU - Chapman, Tim
AU - Wear, Martin A.
AU - Walkinshaw, Malcolm D.
AU - Saxty, Barbara
AU - Melton, David W.
PY - 2015/7/1
Y1 - 2015/7/1
N2 - ERCC1-XPF is a structure-specific endonuclease that is required for the repair of DNA lesions, generated by the widely used platinum-containing cancer chemotherapeutics such as cisplatin, through the Nucleotide Excision Repair and Interstrand Crosslink Repair pathways. Based on mouse xenograft experiments, where ERCC1-deficient melanomas were cured by cisplatin therapy, we proposed that inhibition of ERCC1-XPF could enhance the effectiveness of platinum-based chemotherapy. Here we report the identification and properties of inhibitors against two key targets on ERCC1-XPF. By targeting the ERCC1-XPF interaction domain we proposed that inhibition would disrupt the ERCC1-XPF heterodimer resulting in destabilisation of both proteins. Using in silico screening, we identified an inhibitor that bound to ERCC1-XPF in a biophysical assay, reduced the level of ERCC1-XPF complexes in ovarian cancer cells, inhibited Nucleotide Excision Repair and sensitised melanoma cells to cisplatin. We also utilised high throughput and in silico screening to identify the first reported inhibitors of the other key target, the XPF endonuclease domain. We demonstrate that two of these compounds display specificity in vitro for ERCC1-XPF over two other endonucleases, bind to ERCC1-XPF, inhibit Nucleotide Excision Repair in two independent assays and specifically sensitise Nucleotide Excision Repair-proficient, but not Nucleotide Excision Repair-deficient human and mouse cells to cisplatin.
AB - ERCC1-XPF is a structure-specific endonuclease that is required for the repair of DNA lesions, generated by the widely used platinum-containing cancer chemotherapeutics such as cisplatin, through the Nucleotide Excision Repair and Interstrand Crosslink Repair pathways. Based on mouse xenograft experiments, where ERCC1-deficient melanomas were cured by cisplatin therapy, we proposed that inhibition of ERCC1-XPF could enhance the effectiveness of platinum-based chemotherapy. Here we report the identification and properties of inhibitors against two key targets on ERCC1-XPF. By targeting the ERCC1-XPF interaction domain we proposed that inhibition would disrupt the ERCC1-XPF heterodimer resulting in destabilisation of both proteins. Using in silico screening, we identified an inhibitor that bound to ERCC1-XPF in a biophysical assay, reduced the level of ERCC1-XPF complexes in ovarian cancer cells, inhibited Nucleotide Excision Repair and sensitised melanoma cells to cisplatin. We also utilised high throughput and in silico screening to identify the first reported inhibitors of the other key target, the XPF endonuclease domain. We demonstrate that two of these compounds display specificity in vitro for ERCC1-XPF over two other endonucleases, bind to ERCC1-XPF, inhibit Nucleotide Excision Repair in two independent assays and specifically sensitise Nucleotide Excision Repair-proficient, but not Nucleotide Excision Repair-deficient human and mouse cells to cisplatin.
KW - DNA repair inhibitor
KW - ERCC1-XPF
KW - FANCQ
KW - Interstrand Crosslink Repair
KW - Melanoma
KW - Nucleotide Excision Repair
KW - Ovarian cancer
UR - http://www.scopus.com/inward/record.url?scp=84928794705&partnerID=8YFLogxK
U2 - 10.1016/j.dnarep.2015.04.002
DO - 10.1016/j.dnarep.2015.04.002
M3 - Article
C2 - 25956741
AN - SCOPUS:84928794705
SN - 1568-7864
VL - 31
SP - 19
EP - 28
JO - DNA Repair
JF - DNA Repair
ER -