TY - JOUR
T1 - DNA-PK inhibitor peposertib enhances p53-dependent cytotoxicity of DNA double-strand break inducing therapy in acute leukemia
AU - Haines, Eric
AU - Nishida, Yuki
AU - Carr, Michael I.
AU - Montoya, Rafael Heinz
AU - Ostermann, Lauren B.
AU - Zhang, Weiguo
AU - Zenke, Frank T.
AU - Blaukat, Andree
AU - Andreeff, Michael
AU - Vassilev, Lyubomir T.
N1 - Funding Information:
The work reported in this manuscript was supported in part by Merck KGaA, Darmstadt, Germany (to Lyubomir Vassilev); a UM1 grant (CA186688, Cancer Therapy Evaluation Program, CTEP, National Cancer Institute), the National Institutes of Health Cancer Center Support Grant (P30CA016672), and the Paul and Mary Haas Chair in Genetics (to Michael Andreeff); and Overseas Research Fellowship, Japan Society of the Promotion of Science (to Yuki Nishida). We thank Jazz Pharmaceuticals and Kimberly LeBrun for providing CPX-351 (Vyxeos® ) and Florian Szardenings for his helpful comments.
Funding Information:
Reagents. The DNA-PK inhibitor, M3814, was synthesized at Merck KGaA, Darmstadt, Germany as described. CPX-351 (Vyxeos®) was provided by Jazz Pharmaceuticals as part of a grant from CTEP. Daunorubicin, etoposide, idarubicin, sunitinib, and Z-IETD-FMK were purchased from Selleckchem (Houston, TX). Arabinoside (Cytarabine, AraC) and TRAIL were purchased from MilliporeSigma (Burlington, MA). All inhibitors were dissolved in DMSO to prepare a 10 mM stock and stored at − 20 °C. Aliquots were diluted directly in tissue culture media containing 10% FCS to a desired concentration. The final concentration of DMSO in media did not exceed 0.11% (vol/vol) which has not shown detectable effect on cell morphology, viability and differentiation potential.
Publisher Copyright:
© 2021, The Author(s).
PY - 2021/12
Y1 - 2021/12
N2 - Peposertib (M3814) is a potent and selective DNA-PK inhibitor in early clinical development. It effectively blocks non-homologous end-joining repair of DNA double-strand breaks (DSB) and strongly potentiates the antitumor effect of ionizing radiation (IR) and topoisomerase II inhibitors. By suppressing DNA-PK catalytic activity in the presence of DNA DSB, M3814 potentiates ATM/p53 signaling leading to enhanced p53-dependent antitumor activity in tumor cells. Here, we investigated the therapeutic potential of M3814 in combination with DSB-inducing agents in leukemia cells and a patient-derived tumor. We show that in the presence of IR or topoisomerase II inhibitors, M3814 boosts the ATM/p53 response in acute leukemia cells leading to the elevation of p53 protein levels as well as its transcriptional activity. M3814 synergistically sensitized p53 wild-type, but not p53-deficient, AML cells to killing by DSB-inducing agents via p53-dependent apoptosis involving both intrinsic and extrinsic effector pathways. The antileukemic effect was further potentiated by enhancing daunorubicin-induced myeloid cell differentiation. Further, combined with the fixed-ratio liposomal formulation of daunorubicin and cytarabine, CPX-351, M3814 enhanced the efficacy against leukemia cells in vitro and in vivo without increasing hematopoietic toxicity, suggesting that DNA-PK inhibition could offer a novel clinical strategy for harnessing the anticancer potential of p53 in AML therapy.
AB - Peposertib (M3814) is a potent and selective DNA-PK inhibitor in early clinical development. It effectively blocks non-homologous end-joining repair of DNA double-strand breaks (DSB) and strongly potentiates the antitumor effect of ionizing radiation (IR) and topoisomerase II inhibitors. By suppressing DNA-PK catalytic activity in the presence of DNA DSB, M3814 potentiates ATM/p53 signaling leading to enhanced p53-dependent antitumor activity in tumor cells. Here, we investigated the therapeutic potential of M3814 in combination with DSB-inducing agents in leukemia cells and a patient-derived tumor. We show that in the presence of IR or topoisomerase II inhibitors, M3814 boosts the ATM/p53 response in acute leukemia cells leading to the elevation of p53 protein levels as well as its transcriptional activity. M3814 synergistically sensitized p53 wild-type, but not p53-deficient, AML cells to killing by DSB-inducing agents via p53-dependent apoptosis involving both intrinsic and extrinsic effector pathways. The antileukemic effect was further potentiated by enhancing daunorubicin-induced myeloid cell differentiation. Further, combined with the fixed-ratio liposomal formulation of daunorubicin and cytarabine, CPX-351, M3814 enhanced the efficacy against leukemia cells in vitro and in vivo without increasing hematopoietic toxicity, suggesting that DNA-PK inhibition could offer a novel clinical strategy for harnessing the anticancer potential of p53 in AML therapy.
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U2 - 10.1038/s41598-021-90500-3
DO - 10.1038/s41598-021-90500-3
M3 - Article
C2 - 34108527
AN - SCOPUS:85107423234
SN - 2045-2322
VL - 11
JO - Scientific reports
JF - Scientific reports
IS - 1
M1 - 12148
ER -