title: An evaluation in vitro of PARP-1 inhibitors, rucaparib and olaparib, as radiosensitisers for the treatment of neuroblastoma.
abstract: The radiopharmaceutical (131)I-meta-iodobenzylguanidine ((131)I-MIBG) is an effective treatment for neuroblastoma. However, maximal therapeutic benefit from (131)I-MIBG is likely to be obtained by its combination with chemotherapy. We previously reported enhanced antitumour efficacy of (131)I-MIBG by inhibition of the poly(ADP-ribose) polymerase-1 (PARP-1) DNA repair pathway using the phenanthridinone derivative PJ34. Recently developed alternative PARP-1 inhibitors have greater target specificity and are expected to be associated with reduced toxicity to normal tissue. Therefore, our purpose was to determine whether the more specific PARP-1 inhibitors rucaparib and olaparib enhanced the efficacy of X-radiation or (131)I-MIBG.
  
  
  
  Radiosensitisation of SK-N-BE(2c) neuroblastoma cells or noradrenaline transporter gene-transfected glioma cells (UVW/NAT) was investigated using clonogenic assay. Propidium iodide staining and flow cytometry was used to analyse cell cycle progression. DNA damage was quantified by the phosphorylation of H2AX (γH2AX).
  
  
  
  By combining PARP-1 inhibition with radiation treatment, it was possible to reduce the X-radiation dose or (131)I-MIBG activity concentration required to achieve 50 % cell kill by approximately 50 %. Rucaparib and olaparib were equally effective inhibitors of PARP-1 activity. X-radiation-induced DNA damage was significantly increased 2 h after irradiation by combination with PARP-1 inhibitors (10-fold greater DNA damage compared to untreated controls; p < 0.01). Moreover, combination treatment (i) prevented the restitution of DNA, exemplified by the persistence of 3-fold greater DNA damage after 24 h, compared to untreated controls (p < 0.01) and (ii) induced greater G2/M arrest (p < 0.05) than either single agent alone.
  
  
  
  Rucaparib and olaparib sensitise cancer cells to X-radiation or (131)I-MIBG treatment. It is likely that the mechanism of radiosensitisation entails the accumulation of unrepaired radiation-induced DNA damage. Our findings suggest that the administration of PARP-1 inhibitors and (131)I-MIBG to high risk neuroblastoma patients may be beneficial.
authors: Radiation Oncology, Institute of Cancer Sciences, University of Glasgow, Glasgow, UK. Donna.Nile@glasgow.ac.uk.
authors: Donna L
authors: DL
authors: Nile
authors: Radiation Oncology, Institute of Cancer Sciences, University of Glasgow, Glasgow, UK.
authors: Colin
authors: C
authors: Rae
authors: Radiation Oncology, Institute of Cancer Sciences, University of Glasgow, Glasgow, UK.
authors: Iain J
authors: IJ
authors: Hyndman
authors: University College London Hospitals, London, UK.
authors: Mark N
authors: MN
authors: Gaze
authors: Radiation Oncology, Institute of Cancer Sciences, University of Glasgow, Glasgow, UK.
authors: Robert J
authors: RJ
authors: Mairs
mesh_terms: D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects / Q000528:radiation effects; D059248:Chemoradiotherapy; D004249:DNA Damage; D004260:DNA Repair; D004354:Drug Screening Assays, Antitumor; D006801:Humans; D007211:Indoles / Q000494:pharmacology*; D020128:Inhibitory Concentration 50; D009447:Neuroblastoma / Q000201:enzymology / Q000628:therapy*; D010793:Phthalazines / Q000494:pharmacology*; D010879:Piperazines / Q000494:pharmacology*; D000071137:Poly (ADP-Ribose) Polymerase-1 / Q000037:antagonists & inhibitors / Q000378:metabolism; D000067856:Poly(ADP-ribose) Polymerase Inhibitors / Q000494:pharmacology*; D011838:Radiation-Sensitizing Agents / Q000494:pharmacology*
chemical_list: D007211:Indoles; D010793:Phthalazines; D010879:Piperazines; D000067856:Poly(ADP-ribose) Polymerase Inhibitors; D011838:Radiation-Sensitizing Agents; C531549:rucaparib; C499320:PARP1 protein, human; D000071137:Poly (ADP-Ribose) Polymerase-1; C531550:olaparib
keywords: 131I-MIBG; DNA damage; Neuroblastoma; PARP-1; Targeted radiotherapy