o-(chloroacetylcarbamoyl)fumagillol and Eye-Abnormalities

Previous mode of action studies identified methionine aminopeptidase 2 (MetAP-2) as the target of the antiangiogenic natural product fumagillin and its drug candidate analog, TNP-470. We report here that TNP-470-mediated MetAP-2 inhibition blocks noncanonical Wnt signaling, which plays a critical role in development, cell differentiation, and tumorigenesis. Consistent with this finding, antisense MetAP-2 morpholino oligonucleotide injection in zebrafish embryos phenocopies gastrulation defects seen in noncanonical Wnt5 loss-of-function zebrafish mutants. MetAP-2 inhibition or depletion blocks signaling downstream of the Wnt receptor Frizzled, but upstream of Calmodulin-dependent Kinase II, RhoA, and c-Jun N-terminal Kinase. Moreover, we demonstrate that TNP-470 does not block the canonical Wnt/beta-catenin pathway. Thus, TNP-470 selectively regulates noncanonical over canonical Wnt signaling and provides a unique means to explore and dissect the biological systems mediated by these pathways.

Topics: Adaptor Proteins, Signal Transducing; Aminopeptidases; Angiogenesis Inhibitors; Animals; Calcium-Calmodulin-Dependent Protein Kinase Type 2; Calcium-Calmodulin-Dependent Protein Kinases; Cell Line; Cell Line, Tumor; Cyclohexanes; Dishevelled Proteins; Embryonic Development; Endothelial Cells; Eye Abnormalities; Fatty Acids, Unsaturated; Frizzled Receptors; Humans; JNK Mitogen-Activated Protein Kinases; Metalloendopeptidases; Mice; Molecular Sequence Data; O-(Chloroacetylcarbamoyl)fumagillol; Oligonucleotides, Antisense; Phenotype; Phosphoproteins; Receptors, G-Protein-Coupled; rhoA GTP-Binding Protein; RNA, Small Interfering; Sesquiterpenes; Signal Transduction; Tail; Wnt Proteins; Wnt-5a Protein; Zebrafish; Zebrafish Proteins