title: A Selective Small Molecule DNA2 Inhibitor for Sensitization of Human Cancer Cells to Chemotherapy.
abstract: Cancer cells frequently up-regulate DNA replication and repair proteins such as the multifunctional DNA2 nuclease/helicase, counteracting DNA damage due to replication stress and promoting survival. Therefore, we hypothesized that blocking both DNA replication and repair by inhibiting the bifunctional DNA2 could be a potent strategy to sensitize cancer cells to stresses from radiation or chemotherapeutic agents. We show that homozygous deletion of DNA2 sensitizes cells to ionizing radiation and camptothecin (CPT). Using a virtual high throughput screen, we identify 4-hydroxy-8-nitroquinoline-3-carboxylic acid (C5) as an effective and selective inhibitor of DNA2. Mutagenesis and biochemical analysis define the C5 binding pocket at a DNA-binding motif that is shared by the nuclease and helicase activities, consistent with structural studies that suggest that DNA binding to the helicase domain is necessary for nuclease activity. C5 targets the known functions of DNA2 in vivo: C5 inhibits resection at stalled forks as well as reducing recombination. C5 is an even more potent inhibitor of restart of stalled DNA replication forks and over-resection of nascent DNA in cells defective in replication fork protection, including BRCA2 and BOD1L. C5 sensitizes cells to CPT and synergizes with PARP inhibitors.
authors.affiliation: Colleges of Life Sciences, Zhejiang University, Hangzhou, Zhejiang 310027, China; Department of Cancer Genetics and Epigenetics, Beckman Research Institute, City of Hope, 1500 East Duarte Road, Duarte, CA 91010-3000, USA; Division of Chemistry and Chemical Engineering, Braun Laboratories, California Institute of Technology, Pasadena, CA 91125, USA.
authors.forename: Wenpeng
authors.initials: W
authors.lastname: Liu
authors.affiliation: Department of Cancer Genetics and Epigenetics, Beckman Research Institute, City of Hope, 1500 East Duarte Road, Duarte, CA 91010-3000, USA.
authors.forename: Mian
authors.initials: M
authors.lastname: Zhou
authors.affiliation: Department of Cancer Genetics and Epigenetics, Beckman Research Institute, City of Hope, 1500 East Duarte Road, Duarte, CA 91010-3000, USA.
authors.forename: Zhengke
authors.initials: Z
authors.lastname: Li
authors.affiliation: Molecular Medicine, Beckman Research Institute of City of Hope, 1500 East Duarte Road, Duarte, CA 91010, USA.
authors.forename: Hongzhi
authors.initials: H
authors.lastname: Li
authors.affiliation: Division of Chemistry and Chemical Engineering, Braun Laboratories, California Institute of Technology, Pasadena, CA 91125, USA.
authors.forename: Piotr
authors.initials: P
authors.lastname: Polaczek
authors.affiliation: Department of Cancer Genetics and Epigenetics, Beckman Research Institute, City of Hope, 1500 East Duarte Road, Duarte, CA 91010-3000, USA.
authors.forename: Huifang
authors.initials: H
authors.lastname: Dai
authors.affiliation: Department of Cancer Genetics and Epigenetics, Beckman Research Institute, City of Hope, 1500 East Duarte Road, Duarte, CA 91010-3000, USA.
authors.forename: Qiong
authors.initials: Q
authors.lastname: Wu
authors.affiliation: Colleges of Life Sciences, Zhejiang University, Hangzhou, Zhejiang 310027, China; Department of Cancer Genetics and Epigenetics, Beckman Research Institute, City of Hope, 1500 East Duarte Road, Duarte, CA 91010-3000, USA.
authors.forename: Changwei
authors.initials: C
authors.lastname: Liu
authors.affiliation: Division of Chemistry and Chemical Engineering, Braun Laboratories, California Institute of Technology, Pasadena, CA 91125, USA.
authors.forename: Kenneth K
authors.initials: KK
authors.lastname: Karanja
authors.affiliation: Department of Cancer Biology, University of Texas MD Anderson Cancer Center, Houston, TX 77054, USA.
authors.forename: Vencat
authors.initials: V
authors.lastname: Popuri
authors.affiliation: Division of Chemistry and Chemical Engineering, Braun Laboratories, California Institute of Technology, Pasadena, CA 91125, USA.
authors.forename: Shu-Ou
authors.initials: SO
authors.lastname: Shan
authors.affiliation: Department of Cancer Biology, University of Texas MD Anderson Cancer Center, Houston, TX 77054, USA.
authors.forename: Katharina
authors.initials: K
authors.lastname: Schlacher
authors.affiliation: Department of Cancer Genetics and Epigenetics, Beckman Research Institute, City of Hope, 1500 East Duarte Road, Duarte, CA 91010-3000, USA. Electronic address: lzheng@coh.org.
authors.forename: Li
authors.initials: L
authors.lastname: Zheng
authors.affiliation: Division of Chemistry and Chemical Engineering, Braun Laboratories, California Institute of Technology, Pasadena, CA 91125, USA. Electronic address: jcampbel@caltech.edu.
authors.forename: Judith L
authors.initials: JL
authors.lastname: Campbell
authors.affiliation: Department of Cancer Genetics and Epigenetics, Beckman Research Institute, City of Hope, 1500 East Duarte Road, Duarte, CA 91010-3000, USA. Electronic address: bshen@coh.org.
authors.forename: Binghui
authors.initials: B
authors.lastname: Shen
mesh_terms: D000072283:A549 Cells; D001665:Binding Sites; D002166:Camptothecin / Q000494:pharmacology*; D002264:Carboxylic Acids / Q000737:chemistry / Q000494:pharmacology; D045744:Cell Line, Tumor; D003198:Computer Simulation; D004265:DNA Helicases / Q000037:antagonists & inhibitors / Q000737:chemistry*; D004261:DNA Replication / Q000187:drug effects; D004354:Drug Screening Assays, Antitumor; D004358:Drug Therapy; D004791:Enzyme Inhibitors / Q000737:chemistry / Q000494:pharmacology*; D057166:High-Throughput Screening Assays; D006801:Humans; D061986:MCF-7 Cells; D009369:Neoplasms / Q000188:drug therapy / Q000201:enzymology*; D009600:Nitroquinolines / Q000737:chemistry / Q000494:pharmacology*; D054852:Small Molecule Libraries / Q000737:chemistry / Q000494:pharmacology*
chemical_list: C000614658:4-hydroxy-8-nitroquinoline-3-carboxylic acid; D002264:Carboxylic Acids; D004791:Enzyme Inhibitors; D009600:Nitroquinolines; D054852:Small Molecule Libraries; D004265:DNA Helicases; C509908:DNA2 protein, human; D002166:Camptothecin
keywords: Camptothecin; Cancer; Chemotherapy; DNA binding; DNA end resection; DNA replicatoin fork protection; DNA2 inhibitor; Helicase; Nuclease; PARP inhibitor; Sensitizer