Abstract 1010: A study of combinatorial growth inhibition, cell death and DNA damage repair caused by CHK1 inhibitor SRA737 and WEE1 inhibitor adavosertib in TP53 mutated cell lines
Authors: Adam Stewart, Lisa Pickard, Albert E. Hallsworth, Sylvie Sauvaigo, Giovanna Muggiolu, Florence Raynaud, UDAI BANERJI
Journal: Cancer Research (2021)

Abstract Purpose of study: CHK1 and WEE1 are critical determinants of the G2/M cell cycle checkpoint. The G2/M checkpoint is crucial to enable DNA repair caused by endogenous and exogenous stress. We hypothesised that simultaneous inhibition of CHK1 and WEE1 would cause cell death in TP53 mutated cell lines because of the associated replication stress due to an abrogated G1/S checkpoint. Experimental procedures: The potentiation index of the combination of SRA737 and adavosertib was calculated in six cancer cell lines harbouring TP53 mutations (OVCAR3, COV505, BT20, MDA-MB-436 and CAPAN1, MIAPaCa2) using Cell Titre Blue assays. Cell death and DNA damage was quantified using c-PARP, p-CHK1 Ser345 and gamma H2AX using western blot analysis. In-vivo studies evaluated the efficacy of vehicle control, SRA737, adavosertib and the combination in OVCAR3 and MDA-MB-436 xenograft models. We further explored the mechanisms of DNA damage repair caused by the single agents and the combination using two functional fluorescent multiplex DNA repair assay platforms in OVCAR3 and MDA-MB-436 cells; Exy-SPOT (base excision repair/nucleotide excision repair) and Next-SPOT (double strand break repair). Results: The potentiation index of the addition of SRA737 to adavosertib were 19, 8.6, 8.8, 9.7, 27.2 and 5.7 for COV504, OVCAR3, BT20, MDA-MB-436, CAPAN1 and MIAPaCa2 respectively. The combination of both drugs at single agent GI50 concentrations caused increase in c-PARP, p-CHK1 Ser345 and gamma H2AX in all cell lines studied. We chose to explore the combination in the TP53 mutated OVCAR3 and the TP53 and BRCA1 mutated MDA-MB-436 cell lines. In the OVCAR3 xenograft model, the combination of SRA737 and adavosertib caused tumour regressions and tumour volumes were significantly smaller than controls, p&amp;lt;0001. The combination caused a lesser but significant degree of reduction in the tumour volume in the MDA-MB-436 xenograft model compared to control p=0.003. There was less than a 20% loss of mouse body weight in treatment arms of both xenograft models. Functional DNA damage repair analysis showed the combination of SRA737 and adavosertib caused a significantly greater reduction in oxidative stress related base excision repair (Etheno and 8oxoG lesions) as well as double stranded break repair via alternative end joining in the OVCAR3 cell lines compared to MDA-MB-436 cells, p=0.009, p=0.014 and p=0.005 respectively. Conclusions: The combination of SRA737 and adavosertib caused cell death in TP53 mutated cell lines. There were differences in the DNA damage repair mechanisms in across cell lines and the combination showed significant activity in the xenograft models studied. The combination of SRA737 and adavosertib warrants further evaluation for the treatment of selected TP53 mutated cancers. Citation Format: Adam Stewart, Lisa Pickard, Albert E. Hallsworth, Sylvie Sauvaigo, Giovanna Muggiolu, Florence Raynaud, UDAI BANERJI. A study of combinatorial growth inhibition, cell death and DNA damage repair caused by CHK1 inhibitor SRA737 and WEE1 inhibitor adavosertib in TP53 mutated cell lines [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 1010.