Molecular characterization of endometrial cancer

2010财年生成的初步数据已经确定了子宫内膜癌细胞系亚群中对NF-κ B信号传导的需求。在与G.L.麦克斯韦、CCR、WRAMC的合作中，我们还检查了子宫内膜癌基因表达谱中NF-κ B活性的粗略特征，其在浆液性与类浆液性组织学中平衡，并且在非裔美国人（AA）和高加索人（C）妇女中均匀分布。这种初步的签名是显着较高的癌症从AA妇女，特别是那些浆液性组织学变异。我们实验室正在进行的项目，以及与麦克斯韦实验室的合作，将继续解决既定的目标。具体目的1：鉴定依赖于NF-κ B信号传导的子宫内膜癌的一个亚组。我们通过用IKKb小分子抑制剂处理9种子宫内膜癌细胞系并在XTT测定中定量存活率来测试它们对NF-κ B信号传导的依赖性。我们计划将这些实验扩展到其他细胞系，以及其他功能测定，如粘附，侵袭和细胞因子产生。 特异性目的2：确定子宫内膜癌特异性NF-κ B的基因和蛋白表达特征。我们已经确定了至少2个子宫内膜癌细胞系，在体外对IKKb抑制敏感。我们将分析使用小分子抑制剂和IKKb的shRNA敲低阻断IKKb信号传导后这2种细胞系中的基因表达变化。我们将通过蛋白质印迹证实这些NF-κ B靶点的蛋白质表达降低。这些基因和蛋白质特征将使我们能够使用我们的合作者目前正在收集和生成的基因表达谱和组织微阵列来特异性地询问子宫内膜癌样本。具体目标3：设计诊断和治疗方法，以改善患有这种分子可识别的子宫内膜癌亚型的妇女的临床结果。在未来，我们计划使用目标1和2中产生的数据设计NF-kB靶向药物的临床试验，用于子宫内膜癌显示通路激活证据的妇女。一个潜在的分子靶点是Claudin蛋白，我们发现它在子宫内膜癌中与NF-κ B Rel转录因子共表达。将在临床前研究中检查抗claudin疗法，并且如果有希望，可以在临床环境中探索。

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.