The goal of this UW NCDDG will be to identify novel compounds that demonstrate promise as leads for new anti-cancer therapeutics in preclinical assays. The NCDDG Core B will be responsible for in vitro assays of the compounds produced in Projects 1 and 2 and evaluation of the potency and mechanism of those compounds. Data from Core B will be used by the PI and the steering committee to select: 1)analogs that should be dropped from further analysis, 2) analogs that require further in vitro biological assays and 3) analogs that should be used by Core C and Project 3 for in vivo efficacy testing in autochthonous mouse models of human cancer. In order to efficiently screen many hundreds of new compounds per year in a cost efficient manner, we propose to perform the Core B services at the Keck-UWCCC Small Molecule Screening Facility (http://hts.wisc.edu). The Screening Facility is an existing core service facility for the UW Comprehensive Cancer Center. This facility has the existing infrastructure (cell culture facility, automated liquid handling robots and plate readers), experienced staff and a track record of rapidly screening thousands of chemical compounds in a diverse array of cell-based and biochemical assays. Use of the existing facility will permit faster and cheaper analysis of the NCDDG candidate compounds compared to running those assays in an individual academic laboratory or establishing a new assay facility with NCDDG funds. The goal of Core B is to provide a filter for which compounds move to in vivo testing and not to establish in detail the mechanism of action of large numbers of compound analogs. The Core will use four cell-based assays in 96-well formats that each take advantage of the robotics and experience of the screening facility. Compound analogs will be tested for cytotoxicity in a panel of eight human cancer cell lines obtained from the NCI panel of 60 cell lines. To provide more mechanistic insights into compound function we will focus on two key cellular phenotypes, cell survival and cell migration. Compounds will be assayed for their ability to trigger apoptosis and for their ability to alter the key cell survival mediator Akt. A subset of compounds will also be assayed for their effect on tumor cell migration in vitro. All data will be subject to statistical analysis to ensure the quality of information that is obtained on the new compounds and to facilitate the most informed decisions about the future testing of any compound analog.
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