Abstract 5501: Evaluation of the first potent and highly selective inhibitor of ATR kinase: An approach to selectively sensitize cancer cells to ionizing radiation and hypoxia
Authors: McKenna Gillies , Brunner Thomas , Charlton Peter , Fokas Emmanouil , Hammond Ester , Mangan Matthew , Olcina Monica , Pires Isabel , Pollard John , Prevo Remko , Reaper Philip 
Published: June 2012

DNA damaging agents, such as ionizing radiation (IR), have provided a basis for the treatment of solid tumors for decades, yet they provide only modest benefit for most patients. This reflects, in part, the efficient repair of DNA damage via a complex signaling and repair network known as the DNA damage response (DDR). The ATR, ATM and DNA-PK phosphoinositol 3-kinase-like serine/threonine protein kinases (PIKKs) are the key regulators of the DDR to IR-induced double-strand DNA breaks. This acts to detect the DNA lesions, enforce checkpoints to arrest cell cycle progression, and stimulate repair. Various components of the DDR have been shown to be commonly defective in cancer cells. These cells are widely hypothesized to become dependent on their remaining DDR pathways for survival from DNA damage. The tumor microenvironment has been reported to induce a DDR and in particular, ATR-mediated signaling in response to severe hypoxia.

While inhibitors of a number of DDR enzymes, including ATM, DNA-PK, Chk1 and PARP, have been reported, there have been no reports of drug-like ATR inhibitors. Using the first potent and selective ATR inhibitor (VE-821), we now characterize the effects of ATR inhibition on the cellular response to IR and hypoxia. VE-821 sensitizes many cancer cell lines to IR in both clonogenic and short-term (MTS) assays. For example, VE-821 reduced the surviving fraction of MiaPaca-2 cells treated with 6 Gy IR by 3-fold. Consistent with this, ATR inhibition ablated both RAD51 focus formation and phosphorylation of Chk1 (but not Chk2) after IR treatment, delayed the IR-induced G2/M arrest analyzed by flow cytometry and slowed down DNA replication rates measured using DNA fiber technology. Furthermore, we provide evidence that a basis for the sensitization of cancer cell lines to IR with VE-821 is a synthetic lethal interaction between loss of ATM signaling (a frequent event in cancer resulting from loss of function of proteins such as ATM or p53) and ATR inhibition when cells are treated with IR. In keeping with these results, normal cells (with functional ATM) do not appear sensitized to IR by ATR inhibition. In this case a compensatory DDR is activated, which in turn leads to checkpoint arrest and a strong survival response. Importantly, ATR inhibition also sensitized cancer cells to IR under hypoxic conditions, and to anoxia alone.

These studies show for the first time that a selective ATR inhibitor can preferentially sensitize cancer cells to ionizing radiation under both normoxic and hypoxic conditions and exploit a synthetic lethal interaction between ATM and ATR signaling. This underpins the broad potential of ATR inhibition as a highly promising new strategy to improve the efficacy of DNA damaging therapy.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 5501. doi:10.1158/1538-7445.AM2011-5501