title: PARP inhibitors enhance replication stress and cause mitotic catastrophe in MYCN-dependent neuroblastoma.
abstract: High-risk and MYCN-amplified neuroblastomas are among the most aggressive pediatric tumors. Despite intense multimodality therapies, about 50% of these patients succumb to their disease, making the search for effective therapies an absolute priority. Due to the important functions of poly (ADP-ribose) polymerases, PARP inhibitors have entered the clinical settings for cancer treatment and are being exploited in a variety of preclinical studies and clinical trials. PARP inhibitors based combination schemes have also been tested in neuroblastoma preclinical models with encouraging results. However, the expression of PARP enzymes in human neuroblastoma and the biological consequences of their inhibition remained largely unexplored. Here, we show that high PARP1 and PARP2 expression is significantly associated with high-risk neuroblastoma cases and poor survival, highlighting its previously unrecognized prognostic value for human neuroblastoma. In vitro, PARP1 and 2 are abundant in MYCN amplified and MYCN-overexpressing cells. In this context, PARP inhibitors with high 'PARP trapping' potency, such as olaparib or talazoparib, yield DNA damage and cell death preceded by intense signs of replication stress. Notwithstanding the activation of a CHK1-CDC25A replication stress response, PARP-inhibited MYCN amplified and overexpressing cells fail to sustain a prolonged checkpoint and progress through mitosis in the presence of damaged DNA, eventually undergoing mitotic catastrophe. CHK1-targeted inhibition of the replication stress checkpoint exacerbated this phenotype. These data highlight a novel route for cell death induction by PARP inhibitors and support their introduction, together with CHK1 inhibitors, in therapeutic approaches for neuroblastomas with high MYC(N) activity.
authors.affiliation: Department of Molecular Medicine, University La Sapienza, Rome, Italy.
authors.forename: V
authors.initials: V
authors.lastname: Colicchia
authors.affiliation: Center for Life Nano Science@Sapienza, Istituto Italiano di Tecnologia, Rome, Italy.
authors.forename: M
authors.initials: M
authors.lastname: Petroni
authors.affiliation: Institute of Molecular Biology and Pathology, CNR National Research Council, c/o University La Sapienza, Rome, Italy.
authors.forename: G
authors.initials: G
authors.lastname: Guarguaglini
authors.affiliation: Department of Molecular Medicine, University La Sapienza, Rome, Italy.
authors.forename: F
authors.initials: F
authors.lastname: Sardina
authors.affiliation: Department of Molecular Medicine, University La Sapienza, Rome, Italy.
authors.forename: M
authors.initials: M
authors.lastname: Sahún-Roncero
authors.affiliation: Department of Clinical Medicine, University La Sapienza, Rome, Italy.
authors.forename: M
authors.initials: M
authors.lastname: Carbonari
authors.affiliation: Department of Molecular Medicine, University La Sapienza, Rome, Italy.
authors.forename: B
authors.initials: B
authors.lastname: Ricci
authors.affiliation: Department of Molecular Medicine, University La Sapienza, Rome, Italy.
authors.forename: C
authors.initials: C
authors.lastname: Heil
authors.affiliation: Department of Molecular Medicine, University La Sapienza, Rome, Italy.
authors.forename: C
authors.initials: C
authors.lastname: Capalbo
authors.affiliation: Department of Molecular Medicine, University La Sapienza, Rome, Italy.
authors.forename: F
authors.initials: F
authors.lastname: Belardinilli
authors.affiliation: Department of Experimental Medicine, University La Sapienza, Rome, Italy.
authors.forename: A
authors.initials: A
authors.lastname: Coppa
authors.affiliation: Center for Life Nano Science@Sapienza, Istituto Italiano di Tecnologia, Rome, Italy.
authors.forename: G
authors.initials: G
authors.lastname: Peruzzi
authors.affiliation: Department of Molecular Medicine, University La Sapienza, Rome, Italy.
authors.forename: I
authors.initials: I
authors.lastname: Screpanti
authors.affiliation: Institute of Molecular Biology and Pathology, CNR National Research Council, c/o University La Sapienza, Rome, Italy.
authors.forename: P
authors.initials: P
authors.lastname: Lavia
authors.affiliation: Department of Molecular Medicine, University La Sapienza, Rome, Italy.
authors.forename: A
authors.initials: A
authors.lastname: Gulino
authors.affiliation: Department of Molecular Medicine, University La Sapienza, Rome, Italy.|Istituto Pasteur-Fondazione Cenci Bolognetti, Department of Molecular Medicine, University La Sapienza, Rome, Italy.
authors.forename: G
authors.initials: G
authors.lastname: Giannini
mesh_terms: D017209:Apoptosis / Q000187:drug effects; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D000071877:Checkpoint Kinase 1 / Q000378:metabolism; D002648:Child; D004261:DNA Replication / Q000187:drug effects*; D006801:Humans; D053208:Kaplan-Meier Estimate; D008938:Mitosis / Q000187:drug effects*; D000071447:N-Myc Proto-Oncogene Protein / Q000235:genetics / Q000378:metabolism*; D009447:Neuroblastoma / Q000188:drug therapy*; D000071137:Poly (ADP-Ribose) Polymerase-1 / Q000235:genetics / Q000378:metabolism*; D000067856:Poly(ADP-ribose) Polymerase Inhibitors / Q000008:administration & dosage / Q000627:therapeutic use*; D011065:Poly(ADP-ribose) Polymerases / Q000235:genetics / Q000378:metabolism*
chemical_list: D000071447:N-Myc Proto-Oncogene Protein; 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; D000071877:Checkpoint Kinase 1