Inhibiting WEE1 and IKK-RELA crosstalk overcomes TNFα resistance in head and neck cancers

TNFα is a key mediator of immune and radiotherapy-induced cytotoxicity, but many cancers, including head and neck squamous cell carcinomas (HNSCC), display TNF resistance due to activation of the canonical IKK–NF-κB/RELA pro-survival pathway. However, toxicities associated with direct targeting of the canonical pathway point to the need to identify mechanism(s) contributing to TNFα resistance and synthetic lethal targets to overcome such resistance in cancer cells. Here, RNAi screening for modulators of TNFα–NF-κB reporter activity and cell survival unexpectedly implicated the WEE1 and CDC2 G2–M checkpoint kinases. The IKKα/β-RELA and WEE1-CDC2 signaling pathways are activated by TNFα and form a complex in cell lines derived from both human papillomavirus (−) and (+) subtypes of HNSCC. WEE1 inhibitor AZD1775 reduced IKK/RELA phosphorylation and the expression of NF-κB–dependent pro-survival proteins Cyclin D1 and BCL2. Combination of TNFα and AZD1775 enhanced caspase-mediated apoptosis in vitro, and combination treatment with radiotherapy and AZD1775 potentiated inhibition of HNSCC tumor xenograft growth in vivo, which could be significantly attenuated by TNFα depletion. These data offer new insight into the interplay between NF-κB signaling and WEE1-mediated regulation of the G2–M cell-cycle checkpoint in HNSCC. Implications Inhibiting WEE1 and IKK-RELA crosstalk could potentially enhance the effects of therapies mediated by TNFα with less systemic immune suppression and toxicity than observed with direct interruption of IKK-NF-κB/RELA signaling

Additional file 1: of Integrated transcriptional profiling and genomic analyses reveal RPN2 and HMGB1 as promising biomarkers in colorectal cancer

Table S1. List of differentially expressed genes with transcriptional regulation. Table S2. Gene Ontology (GO) and Ingenuity Pathway Analysis (IPA) of differentially expressed genes based on biological processes, canonical pathways and upstream regulators. a GO biological processes enriched (P<0.01, FDR<5%). b Upstream regulator analysis using Ingenuity Pathway Analysis 2013 (P<0.00001) Table S3. Comparison of differentially expressed genes with published CRC microarray studie

Additional file 2: of Integrated transcriptional profiling and genomic analyses reveal RPN2 and HMGB1 as promising biomarkers in colorectal cancer

Figure S1. Ingenuity Pathway Analysis (IPA) identifies enriched top pathway networks in CRC.Figure S2. Oncoprint summary of genomic alterations in CRC from the TCGA database. Figure S3. Clininopathological characteristics of CRC sample