Molecular characterization of endometrial cancer

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Preliminary data generated in FY2010 have established the requirement for NF-kB signaling in a subset of endometrial cancer cell lines. In collaboration with G.L.Maxwell, CCR, WRAMC, we have also examined a rough signature of NF-kB activity in the gene expression profiles of endometrial cancers balanced for serous vs endometrioid histology and also distributed equally among African American (AA) and caucasian (C) women. This preliminary signature is significantly higher in cancers from AA women, especially those of the serous histology variant. Ongoing projects in our lab, and in collaboration with the Maxwell lab, will continue to address the stated goals. SPECIFIC AIM 1: to identify a subset of endometrial cancers that are dependent on NF-kB signaling. We have tested 9 endometrial cancer cell lines for their dependence on NF-kB signaling by treating them with IKKb small molecule inhibitor and quantifying viability in XTT assays. We plan to extend these experiments to additional cell lines, and in additional function assays such as adhesion, invasion and cytokine production. SPECIFIC AIM 2: to define gene and protein expression signatures of NF-kB specific to endometrial cancer. We have identified at least 2 endometrial cancer cell lines that are sensitive to IKKb inhibition in vitro. We will profile gene expression changes in these 2 cell lines upon blockade of IKKb signaling using both a small molecule inhibitor and shRNA knockdown of IKKb. We will confirm decrease in protein expression of these NF-kB targets by Western blot. These gene and protein signatures will allow us to specifically interrogate the endometrial cancer samples using gene expression profiles and tissue microarrays that are currently being collected and generated by our collaborator. SPECIFIC AIM 3: to design diagnostics and therapeutics that will improve the clinical outcome of women with this molecularly identifiable subtype of endometrial cancer. In the future, we plan to use the data generated in Aims 1 and 2 to design clinical trials with NF-kB targeted agents for women whose endometrial cancers show evidence of pathway activation. One potential molecular target is the Claudin protein, which we have found to be co-expressed with NF-kB Rel transcription factors in endometrial cancers. Anti-claudin therapeutics will be examined in preclinical studies, and if promising may be explored in the clincal setting.