title: Inhibition of Pax2 Transcription Activation with a Small Molecule that Targets the DNA Binding Domain.
abstract: The Pax gene family encodes DNA binding transcription factors that control critical steps in embryonic development and differentiation of specific cell lineages. Often, Pax proteins are re-expressed or ectopically expressed in cancer and other diseases of abnormal proliferation, making them attractive targets for tissue specific inhibition by small molecules. In this report, we used a homology model of the Pax2 paired domain and a virtual screen to identify small molecules that can inhibit binding of the paired domain to DNA and Pax2 mediated transcription activation. Candidates from the virtual screen were then confirmed in a cell based Pax2 transactivation assay. Subsequently, we tested analogs of these hits to identify a single compound that effectively blocked Pax2 activity and DNA binding with a Kdof 1.35-1.5 μM. The compound, termed EG1, was used to inhibit embryonic kidney development, a process directly dependent on Pax2 activity. Furthermore, we show that EG1 can inhibit proliferation of Pax2 positive renal and ovarian cancer cell lines but has little effect on Pax2 negative cancer cells. These data confirm that small molecules targeting the DNA binding paired domain can be identified and may be good lead compounds for developing tissue and cell-type specific anticancer therapies.
authors.affiliation: Department of Pathology, University of Michigan , Ann Arbor, Michigan 48109, United States.|Molecular and Cellular Pathology Graduate Program, University of Michigan , Ann Arbor, Michigan 48109, United States.
authors.forename: Edward
authors.identifier: 0000-0003-0438-506X
authors.initials: E
authors.lastname: Grimley
authors.affiliation: Department of Pathology, University of Michigan , Ann Arbor, Michigan 48109, United States.
authors.forename: Chenzhong
authors.initials: C
authors.lastname: Liao
authors.affiliation: Department of Pathology, University of Michigan , Ann Arbor, Michigan 48109, United States.
authors.forename: Egon J
authors.initials: EJ
authors.lastname: Ranghini
authors.affiliation: Department of Pathology, University of Michigan , Ann Arbor, Michigan 48109, United States.
authors.forename: Zaneta
authors.initials: Z
authors.lastname: Nikolovska-Coleska
authors.affiliation: Department of Pathology, University of Michigan , Ann Arbor, Michigan 48109, United States.
authors.forename: Gregory R
authors.identifier: 0000-0002-7967-6104
authors.initials: GR
authors.lastname: Dressler
mesh_terms: D000818:Animals; D045744:Cell Line, Tumor; D003198:Computer Simulation; D004247:DNA / Q000378:metabolism*; D057809:HEK293 Cells; D006801:Humans; D007668:Kidney / Q000254:growth & development / Q000378:metabolism; D051762:PAX2 Transcription Factor / Q000037:antagonists & inhibitors / Q000378:metabolism*; D054852:Small Molecule Libraries; D015533:Transcriptional Activation
chemical_list: D051762:PAX2 Transcription Factor; D054852:Small Molecule Libraries; D004247:DNA