title: Mechanism of radiosensitization by the Chk1/2 inhibitor AZD7762 involves abrogation of the G2 checkpoint and inhibition of homologous recombinational DNA repair.
abstract: The median survival for patients with locally advanced pancreatic cancer treated with gemcitabine and radiation is approximately 1 year. To develop improved treatment, we have combined a Chk1/2-targeted agent, AZD7762, currently in phase I clinical trials, with gemcitabine and ionizing radiation in preclinical pancreatic tumor models. We found that in vitro AZD7762 alone or in combination with gemcitabine significantly sensitized MiaPaCa-2 cells to radiation. AZD7762 inhibited Chk1 autophosphorylation (S296 Chk1), stabilized Cdc25A, and increased ATR/ATM-mediated Chk1 phosphorylation (S345 Chk1). Radiosensitization by AZD7762 was associated with abrogation of the G(2) checkpoint as well as with inhibition of Rad51 focus formation, inhibition of homologous recombination repair, and persistent gamma-H2AX expression. AZD7762 was also a radiation sensitizer in multiple tumor xenograft models. In both MiaPaCa-2- and patient-derived xenografts, AZD7762 significantly prolonged the median time required for tumor volume doubling in response to gemcitabine and radiation. Together, our findings suggest that G(2) checkpoint abrogation and homologous recombination repair inhibition both contribute to sensitization by Chk1 inhibition. Furthermore, they support the clinical use of AZD7762 in combination with gemcitabine and radiation for patients with locally advanced pancreatic cancer.
authors.affiliation: Department of Radiation Oncology, University of Michigan Medical School, Ann Arbor, Michigan 48109-5637, USA. mmccrack@med.umich.edu
authors.forename: Meredith A
authors.initials: MA
authors.lastname: Morgan
authors.forename: Leslie A
authors.initials: LA
authors.lastname: Parsels
authors.forename: Lili
authors.initials: L
authors.lastname: Zhao
authors.forename: Joshua D
authors.initials: JD
authors.lastname: Parsels
authors.forename: Mary A
authors.initials: MA
authors.lastname: Davis
authors.forename: Maria C
authors.initials: MC
authors.lastname: Hassan
authors.forename: Sankari
authors.initials: S
authors.lastname: Arumugarajah
authors.forename: Linda
authors.initials: L
authors.lastname: Hylander-Gans
authors.forename: Deborah
authors.initials: D
authors.lastname: Morosini
authors.forename: Diane M
authors.initials: DM
authors.lastname: Simeone
authors.forename: Christine E
authors.initials: CE
authors.lastname: Canman
authors.forename: Daniel P
authors.initials: DP
authors.lastname: Normolle
authors.forename: Sonya D
authors.initials: SD
authors.lastname: Zabludoff
authors.forename: Jonathan
authors.initials: J
authors.lastname: Maybaum
authors.forename: Theodore S
authors.initials: TS
authors.lastname: Lawrence
mesh_terms: D000230:Adenocarcinoma / Q000188:drug therapy / Q000378:metabolism / Q000473:pathology; D000818:Animals; D015153:Blotting, Western; D045744:Cell Line, Tumor; D000071877:Checkpoint Kinase 1; D064447:Checkpoint Kinase 2; D004249:DNA Damage / Q000187:drug effects / Q000528:radiation effects; D004260:DNA Repair / Q000187:drug effects* / Q000528:radiation effects; D003841:Deoxycytidine / Q000031:analogs & derivatives / Q000494:pharmacology; D004359:Drug Therapy, Combination; D005434:Flow Cytometry; D005455:Fluorescent Antibody Technique; D016195:G2 Phase / Q000187:drug effects* / Q000528:radiation effects; D005720:Gamma Rays; D006801:Humans; D015151:Immunoblotting; D007124:Immunoenzyme Techniques; D051379:Mice; D016688:Mice, Inbred NOD; D016513:Mice, SCID; D010190:Pancreatic Neoplasms / Q000378:metabolism / Q000473:pathology / Q000532:radiotherapy*; D047428:Protein Kinase Inhibitors / Q000494:pharmacology*; D011494:Protein Kinases / Q000737:chemistry / Q000378:metabolism; D017346:Protein Serine-Threonine Kinases / Q000037:antagonists & inhibitors*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D051135:Rad51 Recombinase / Q000378:metabolism; D011838:Radiation-Sensitizing Agents / Q000494:pharmacology*; D011995:Recombination, Genetic / Q000187:drug effects / Q000528:radiation effects; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013876:Thiophenes / Q000494:pharmacology*; D014508:Urea / Q000031:analogs & derivatives* / Q000494:pharmacology; D023041:Xenograft Model Antitumor Assays; D000093542:Gemcitabine
chemical_list: C532363:3-(carbamoylamino)-5-(3-fluorophenyl)-N-(3-piperidyl)thiophene-2-carboxamide; D047428:Protein Kinase Inhibitors; D012333:RNA, Messenger; D011838:Radiation-Sensitizing Agents; D013876:Thiophenes; D003841:Deoxycytidine; D014508:Urea; D011494:Protein Kinases; D064447:Checkpoint Kinase 2; C000605867:CHEK1 protein, human; C578634:CHEK2 protein, human; D000071877:Checkpoint Kinase 1; C000605869:Chek1 protein, mouse; C578633:Chek2 protein, mouse; D017346:Protein Serine-Threonine Kinases; D051135:Rad51 Recombinase; D000093542:Gemcitabine