title: Wee1 inhibition potentiates Wip1-dependent p53-negative tumor cell death during chemotherapy.
abstract: Inactivation of p53 found in more than half of human cancers is often associated with increased tumor resistance to anti-cancer therapy. We have previously shown that overexpression of the phosphatase Wip1 in p53-negative tumors sensitizes them to chemotherapeutic agents, while protecting normal tissues from the side effects of anti-cancer treatment. In this study, we decided to search for kinases that prevent Wip1-mediated sensitization of cancer cells, thereby interfering with efficacy of genotoxic anti-cancer drugs. To this end, we performed a flow cytometry-based screening in order to identify kinases that regulated the levels of γH2AX, which were used as readout. Another criterion of the screen was increased sensitivity of p53-negative tumor cells to cisplatin (CDDP) in a Wip1-dependent manner. We have found that a treatment with a low dose (75 nM) of MK-1775, a recently described specific chemical inhibitor of Wee1, decreases CDDP-induced H2AX phosphorylation in p53-negative cells and enhances the Wip1-sensitization of p53-negative tumors. We were able to reduce CDDP effective concentration by 40% with a combination of Wip1 overexpression and Wee1 kinase inhibition. We have observed that Wee1 inhibition potentiates Wip1-dependent tumor sensitization effect by reducing levels of Hipk2 kinase, a negative regulator of Wip1 pathway. In addition, during CDDP treatment, the combination of Wee1 inhibition and Wip1 overexpression has a mild but significant protective effect in normal cells and tissues. Our results indicate that inhibition of the negative regulators of Wip1 pathway, Wee1 and Hipk2, in p53-negative tumors could potentiate efficiency of chemotherapeutic agents without concomitant increase of cytotoxicity in normal tissues. The development and clinical use of Wee1 and Hipk1 kinase chemical inhibitors might be a promising strategy to improve anti-cancer therapy.
authors.affiliation: INSERM UMR 866, Laboratoire d'excellence ARC, Dijon, France.|University of Burgundy, Dijon, France.
authors.forename: V
authors.initials: V
authors.lastname: Clausse
authors.affiliation: INSERM UMR 866, Laboratoire d'excellence ARC, Dijon, France.|University of Burgundy, Dijon, France.
authors.forename: A R
authors.initials: AR
authors.lastname: Goloudina
authors.affiliation: INSERM UMR 866, Laboratoire d'excellence ARC, Dijon, France.|University of Burgundy, Dijon, France.
authors.forename: B
authors.initials: B
authors.lastname: Uyanik
authors.affiliation: Institute of Cytology, RAS, St. Petersburg, Russia.
authors.forename: E Y
authors.initials: EY
authors.lastname: Kochetkova
authors.affiliation: INSERM UMR 866, Laboratoire d'excellence ARC, Dijon, France.|University of Burgundy, Dijon, France.
authors.forename: S
authors.initials: S
authors.lastname: Richaud
authors.affiliation: Institute of Cytology, RAS, St. Petersburg, Russia.
authors.forename: O A
authors.initials: OA
authors.lastname: Fedorova
authors.affiliation: INSERM UMR 866, Laboratoire d'excellence ARC, Dijon, France.|University of Burgundy, Dijon, France.
authors.forename: A
authors.initials: A
authors.lastname: Hammann
authors.affiliation: INSERM UMR 866, Laboratoire d'excellence ARC, Dijon, France.|University of Burgundy, Dijon, France.
authors.forename: M
authors.initials: M
authors.lastname: Bardou
authors.affiliation: Institute of Cytology, RAS, St. Petersburg, Russia.
authors.forename: N A
authors.initials: NA
authors.lastname: Barlev
authors.affiliation: INSERM UMR 866, Laboratoire d'excellence ARC, Dijon, France.|University of Burgundy, Dijon, France.|Anticancer Center Georges François Leclerc, Dijon, France.
authors.forename: C
authors.initials: C
authors.lastname: Garrido
authors.affiliation: INSERM UMR 866, Laboratoire d'excellence ARC, Dijon, France.|University of Burgundy, Dijon, France.|Institute of Cytology, RAS, St. Petersburg, Russia.
authors.forename: O N
authors.initials: ON
authors.lastname: Demidov
mesh_terms: D000818:Animals; D000970:Antineoplastic Agents / Q000494:pharmacology / Q000627:therapeutic use; D017209:Apoptosis / Q000187:drug effects; D002352:Carrier Proteins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D053148:Caspase 3 / Q000378:metabolism; D018797:Cell Cycle Proteins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D045744:Cell Line, Tumor; D002945:Cisplatin / Q000494:pharmacology / Q000627:therapeutic use; D015179:Colorectal Neoplasms / Q000188:drug therapy / Q000401:mortality / Q000473:pathology; D004249:DNA Damage / Q000187:drug effects; D059565:G2 Phase Cell Cycle Checkpoints / Q000187:drug effects; D006657:Histones / Q000378:metabolism; D006801:Humans; D051379:Mice; D008822:Mice, Transgenic; D008928:Mitochondria / Q000378:metabolism; D009687:Nuclear Proteins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D010766:Phosphorylation / Q000187:drug effects; D000071636:Protein Phosphatase 2C / Q000235:genetics / Q000378:metabolism*; D017346:Protein Serine-Threonine Kinases / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D011505:Protein-Tyrosine Kinases / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D034622:RNA Interference; D015996:Survival Rate; D016159:Tumor Suppressor Protein p53 / Q000172:deficiency / Q000235:genetics*
chemical_list: D000970:Antineoplastic Agents; D002352:Carrier Proteins; D018797:Cell Cycle Proteins; C468783:H2AX protein, human; D006657:Histones; D009687:Nuclear Proteins; D016159:Tumor Suppressor Protein p53; C115114:HIPK2 protein, human; D011505:Protein-Tyrosine Kinases; C499349:WEE1 protein, human; D017346:Protein Serine-Threonine Kinases; C000607458:PPM1D protein, human; D000071636:Protein Phosphatase 2C; D053148:Caspase 3; D002945:Cisplatin