title: Combined inhibition of Wee1 and PARP1/2 for radiosensitization in pancreatic cancer.
abstract: OBJECTIVE
  While the addition of radiation to chemotherapy improves survival in patients with locally advanced pancreatic cancer, more effective therapies are urgently needed. Thus, we investigated the radiosensitizing efficacy of the novel drug combination of Wee1 and PARP1/2 inhibitors (AZD1775 and olaparib, respectively) in pancreatic cancer.
  
  
  METHODS
  Radiosensitization of AsPC-1 or MiaPaCa-2 human pancreatic cancer cells was assessed by clonogenic survival and tumor growth assays. Mechanistically, the effects of AZD1775, olaparib, and radiation on cell cycle, DNA damage (γH2AX), and homologous recombination repair (HRR) were determined.
  
  
  RESULTS
  Treatment of AsPC-1 and MiaPaCa-2 cells with either AZD1775 or olaparib caused modest radiosensitization, whereas treatment with the combination significantly increased radiosensitization. Radiosensitization by the combination of AZD1775 and olaparib was associated with G2 checkpoint abrogation and persistent DNA damage. In addition, AZD1775 inhibited HRR activity and prevented radiation-induced Rad51 focus formation. Finally, in vivo, in MiaPaCa-2-derived xenografts, olaparib did not radiosensitize, whereas AZD1775 produced moderate, yet significant, radiosensitization (P < 0.05). Importantly, the combination of AZD1775 and olaparib produced highly significant radiosensitization (P < 0.0001) evidenced by a 13-day delay in tumor volume doubling (vs. radiation alone) and complete eradication of 20% of tumors.
  
  
  CONCLUSIONS
  Taken together, these results demonstrate the efficacy of combined inhibition of Wee1 and PARP inhibitors for radiosensitizing pancreatic cancers and support the model that Wee1 inhibition sensitizes cells to PARP inhibitor-mediated radiosensitization through inhibition of HRR and abrogation of the G2 checkpoint, ultimately resulting in unrepaired, lethal DNA damage and radiosensitization. Clin Cancer Res; 20(19); 5085-96. ©2014 AACR.
authors.affiliation: Department of Radiation Oncology, University of Michigan Medical School, Ann Arbor, Michigan.
authors.forename: David
authors.initials: D
authors.lastname: Karnak
authors.affiliation: Department of Radiation Oncology, University of Michigan Medical School, Ann Arbor, Michigan.
authors.forename: Carl G
authors.initials: CG
authors.lastname: Engelke
authors.affiliation: Department of Pharmacology, University of Michigan Medical School, Ann Arbor, Michigan.
authors.forename: Leslie A
authors.initials: LA
authors.lastname: Parsels
authors.affiliation: Department of Radiation Oncology, University of Michigan Medical School, Ann Arbor, Michigan.
authors.forename: Tasneem
authors.initials: T
authors.lastname: Kausar
authors.affiliation: Department of Radiation Oncology, University of Michigan Medical School, Ann Arbor, Michigan.
authors.forename: Dongping
authors.initials: D
authors.lastname: Wei
authors.affiliation: Department of Radiation Oncology, University of Michigan Medical School, Ann Arbor, Michigan.
authors.forename: Jordan R
authors.initials: JR
authors.lastname: Robertson
authors.affiliation: Department of Radiation Oncology, University of Michigan Medical School, Ann Arbor, Michigan.
authors.forename: Katherine B
authors.initials: KB
authors.lastname: Marsh
authors.affiliation: Department of Radiation Oncology, University of Michigan Medical School, Ann Arbor, Michigan.
authors.forename: Mary A
authors.initials: MA
authors.lastname: Davis
authors.affiliation: Biostatistics Unit, University of Michigan Comprehensive Cancer Center, Ann Arbor, Michigan.
authors.forename: Lili
authors.initials: L
authors.lastname: Zhao
authors.affiliation: Department of Pharmacology, University of Michigan Medical School, Ann Arbor, Michigan.
authors.forename: Jonathan
authors.initials: J
authors.lastname: Maybaum
authors.affiliation: Department of Radiation Oncology, University of Michigan Medical School, Ann Arbor, Michigan.
authors.forename: Theodore S
authors.initials: TS
authors.lastname: Lawrence
authors.affiliation: Department of Radiation Oncology, University of Michigan Medical School, Ann Arbor, Michigan. mmccrack@med.umich.edu.
authors.forename: Meredith A
authors.initials: MA
authors.lastname: Morgan
mesh_terms: D000818:Animals; D018797:Cell Cycle Proteins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects / Q000528:radiation effects; D004249:DNA Damage / Q000187:drug effects / Q000528:radiation effects; D004195:Disease Models, Animal; D005260:Female; D006801:Humans; D009687:Nuclear Proteins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D010190:Pancreatic Neoplasms / Q000235:genetics / Q000378:metabolism* / Q000473:pathology; D010793:Phthalazines / Q000494:pharmacology; D010879:Piperazines / Q000494:pharmacology; D000071137:Poly (ADP-Ribose) Polymerase-1; D000067856:Poly(ADP-ribose) Polymerase Inhibitors; D011065:Poly(ADP-ribose) Polymerases / Q000235:genetics / Q000378:metabolism*; D011505:Protein-Tyrosine Kinases / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D011827:Radiation; D011836:Radiation Tolerance / Q000187:drug effects / Q000235:genetics; D047368:Tumor Burden / Q000187:drug effects / Q000235:genetics / Q000528:radiation effects; D023041:Xenograft Model Antitumor Assays
chemical_list: D018797:Cell Cycle Proteins; D009687:Nuclear Proteins; D010793:Phthalazines; D010879:Piperazines; D000067856:Poly(ADP-ribose) Polymerase Inhibitors; C499320:PARP1 protein, human; C483041:PARP2 protein, human; D000071137:Poly (ADP-Ribose) Polymerase-1; D011065:Poly(ADP-ribose) Polymerases; D011505:Protein-Tyrosine Kinases; C499349:WEE1 protein, human; C531550:olaparib