title: Assessment of chk1 phosphorylation as a pharmacodynamic biomarker of chk1 inhibition.
abstract: OBJECTIVE
  Chk1 inhibitors, such as AZD7762, are in clinical development in combination with cytotoxic agents for the treatment of solid tumors, including pancreatic cancers. To maximize the likelihood of their clinical success, it is essential to optimize drug scheduling as well as pharmacodynamic biomarkers in preclinical models.
  
  
  METHODS
  We tested multiple schedules of administration of gemcitabine and AZD7762 on the survival of pancreatic cancer cells. Potential pharmacodynamic biomarkers including pChk1, pChk2, pHistone H3, and caspase-3 were evaluated in vitro, followed by assessment of promising candidate biomarkers in vivo. We then went on to determine the contributions of PP2A and DNA damage to the mechanism(s) of induction of the identified biomarker, pS345 Chk1.
  
  
  RESULTS
  AZD7762 given during and after or after gemcitabine administration produced maximum chemosensitization. In vivo, AZD7762 significantly inhibited the growth of pancreatic tumor xenografts in response to gemcitabine. Of the biomarkers assessed, pS345 Chk1 was most consistently increased in response to gemcitabine and AZD7762 in tumors and normal tissues (hair follicles). pS345 Chk1 induction in response to gemcitabine and AZD7762 occurred in the presence of PP2A inhibition and in association with elevated γH2AX, suggesting that DNA damage is an underlying mechanism.
  
  
  CONCLUSIONS
  AZD7762 sensitizes pancreatic cancer cells and tumors to gemcitabine in association with induction of pS345 Chk1. Together these data support the clinical investigation of AZD7762 with gemcitabine in pancreatic cancer under a dosing schedule in which gemcitabine is administered concurrent with or before AZD7762 and in conjunction with skin biopsies to measure pS345 Chk1.
authors.affiliation: Departments of Radiation Oncology, Pharmacology, and Surgery, University of Michigan Medical School, Ann Arbor, MI 48109-5637, USA.
authors.forename: Leslie A
authors.initials: LA
authors.lastname: Parsels
authors.forename: Yushen
authors.initials: Y
authors.lastname: Qian
authors.forename: Daria M
authors.initials: DM
authors.lastname: Tanska
authors.forename: Marisa
authors.initials: M
authors.lastname: Gross
authors.forename: Lili
authors.initials: L
authors.lastname: Zhao
authors.forename: Maria C
authors.initials: MC
authors.lastname: Hassan
authors.forename: Sankari
authors.initials: S
authors.lastname: Arumugarajah
authors.forename: Joshua D
authors.initials: JD
authors.lastname: Parsels
authors.forename: Linda
authors.initials: L
authors.lastname: Hylander-Gans
authors.forename: Diane M
authors.initials: DM
authors.lastname: Simeone
authors.forename: Deborah
authors.initials: D
authors.lastname: Morosini
authors.forename: Jeffrey L
authors.initials: JL
authors.lastname: Brown
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
authors.forename: Meredith A
authors.initials: MA
authors.lastname: Morgan
mesh_terms: D000818:Animals; D015415:Biomarkers / Q000378:metabolism; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D000071877:Checkpoint Kinase 1; D004249:DNA Damage; D003841:Deoxycytidine / Q000031:analogs & derivatives* / Q000494:pharmacology; D006657:Histones / Q000378:metabolism; D006801:Humans; D051379:Mice; D010190:Pancreatic Neoplasms / Q000188:drug therapy; D010766:Phosphorylation / Q000187:drug effects; D047428:Protein Kinase Inhibitors / Q000494:pharmacology*; D011494:Protein Kinases / Q000378:metabolism*; D013876:Thiophenes / Q000494:pharmacology*; D014183:Transplantation, Heterologous; 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; D015415:Biomarkers; D006657:Histones; D047428:Protein Kinase Inhibitors; D013876:Thiophenes; D003841:Deoxycytidine; D014508:Urea; D011494:Protein Kinases; C000605867:CHEK1 protein, human; D000071877:Checkpoint Kinase 1; C000605869:Chek1 protein, mouse; D000093542:Gemcitabine