title: High-throughput screening identifies novel agents eliciting hypersensitivity in Fanconi pathway-deficient cancer cells.
abstract: Inactivation of the Fanconi anemia (FA) pathway occurs in diverse human tumors among the general population and renders those tumors hypersensitive to DNA interstrand-cross-linking (ICL) agents. The identification of novel agents to which FA pathway-deficient cells were hypersensitive could provide new therapeutic opportunities and improve our molecular understanding of the FA genes. Using high-throughput screening, we assessed the growth of isogenic human cancer cells that differed only in the presence or absence of single FA genes upon treatment with 880 active drugs and 40,000 diverse compounds. We identified several compounds to which FA pathway-deficient cells were more sensitive than FA pathway-proficient cells, including two groups of structurally related compounds. We further investigated the compound eliciting the strongest effect, termed 80136342. Its mechanism of action was distinct from that of ICL agents; 80136342 did not cause increased chromosomal aberrations, enhanced FANCD2 monoubiquitination, H2AX phosphorylation, p53 activation, or ICL induction. Similar to ICL agents, however, 80136342 caused a pronounced G(2) arrest in FA pathway-deficient cells. When applied in combination with ICL agents, 80136342 had at least additive toxic effects, excluding interferences on ICL-induced toxicity and facilitating a combinational application. Finally, we identified one particular methyl group necessary for the effects of 80136342 on FA-deficient cells. In conclusion, using high-throughput screening in an isogenic human FA cancer model, we explored a novel approach to identify agents eliciting hypersensitivity in FA pathway-deficient cells. We discovered several attractive candidates to serve as lead compounds for evaluating structure-activity relationships and developing therapeutics selectively targeting FA pathway-deficient tumors.
authors.affiliation: The Sidney Kimmel Comprehensive Cancer Center at Johns Hopkins University, 1650 Orleans Street, Baltimore, MD 21231, USA.
authors.forename: Eike
authors.initials: E
authors.lastname: Gallmeier
authors.forename: Tomas
authors.initials: T
authors.lastname: Hucl
authors.forename: Jonathan R
authors.initials: JR
authors.lastname: Brody
authors.forename: David A
authors.initials: DA
authors.lastname: Dezentje
authors.forename: Khola
authors.initials: K
authors.lastname: Tahir
authors.forename: Jana
authors.initials: J
authors.lastname: Kasparkova
authors.forename: Viktor
authors.initials: V
authors.lastname: Brabec
authors.forename: Kurtis E
authors.initials: KE
authors.lastname: Bachman
authors.forename: Scott E
authors.initials: SE
authors.lastname: Kern
mesh_terms: D000970:Antineoplastic Agents / Q000008:administration & dosage / Q000032:analysis*; D000971:Antineoplastic Combined Chemotherapy Protocols / Q000627:therapeutic use; D019457:Chromosome Breakage / Q000187:drug effects; D003432:Cross-Linking Reagents / Q000494:pharmacology; D004353:Drug Evaluation, Preclinical; D004354:Drug Screening Assays, Antitumor; D051856:Fanconi Anemia Complementation Group Proteins / Q000235:genetics*; D006657:Histones / Q000378:metabolism; D006801:Humans; D008954:Models, Biological; D009369:Neoplasms / Q000188:drug therapy / Q000235:genetics*; D011725:Pyridines / Q000627:therapeutic use; D011804:Quinolines / Q000627:therapeutic use; D015398:Signal Transduction / Q000187:drug effects*; D014407:Tumor Cells, Cultured
chemical_list: C518743:1,2,3,5,7,9-hexamethyl-3H-pyrrolo(3,2-f)quinoline; D000970:Antineoplastic Agents; D003432:Cross-Linking Reagents; D051856:Fanconi Anemia Complementation Group Proteins; C468783:H2AX protein, human; D006657:Histones; D011725:Pyridines; D011804:Quinolines