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Mechanism of action of selective inhibitors of IL-6 induced STAT3 pathway in head and neck cancer cell lines

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Journal of Chemical Biology

Abstract

Studies indicate that elevated interleukin-6 (IL-6) levels engage IL6Rα-gp130 receptor complexes to activate signal transducer and activator of transcription 3 (STAT3) that is hyperactivated in many cancers including head and neck squamous cell carcinoma (HNSCC). Our previous HCS campaign identified several hits that selectively blocked IL-6-induced STAT3 activation. This study describes our investigation of the mechanism(s) of action of three of the four chemical series that progressed to lead activities: a triazolothiadiazine (864669), amino alcohol (856350), and an oxazole-piperazine (4248543). We demonstrated that all three blocked IL-6-induced upregulation of the cyclin D1 and Bcl-XL STAT3 target genes. None of the compounds exhibited direct binding interactions with STAT3 in surface plasmon resonance (SPR) binding assays; neither did they inhibit the recruitment and binding of a phospho-tyrosine-gp130 peptide to STAT3 in a fluorescence polarization assay. Furthermore, they exhibited little or no inhibition in a panel of 83 cancer-associated in vitro kinase profiling assays, including lack of inhibition of IL-6-induced Janus kinase (JAK 1, 2, and 3) activation. Further, 864669 and 4248543 selectively inhibited IL-6-induced STAT3 activation but not that induced by oncostatin M (OSM). The compounds 864669 and 4248543 abrogated IL-6-induced phosphorylation of the gp130 signaling subunit (phospho-gp130Y905) of the IL-6-receptor complex in HNSCC cell lines which generate docking sites for the SH2 domains of STAT3. Our data indicate that 864669 and 4248543 block IL-6-induced STAT activation by interfering with the recruitment, assembly, or activation of the hexamer-activated IL-6/IL-6Rα/gp130 signaling complex that occurs after IL-6 binding to IL-6Rα subunits.

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Authors and Affiliations

  1. Department of Otolaryngology, University of Pittsburgh, Pittsburgh, PA, 15213, USA

    Malabika Sen, Netanya I. Pollock, Kara DeGrave, Sonali C. Joyce & Maria L. Freilino

  2. Department of Pharmaceutical Sciences, University of Pittsburgh, Pittsburgh, PA, 15213, USA

    Paul A. Johnston, Yun Hua, Daniel P. Camarco & David A. Close

  3. University of Pittsburgh Chemical Diversity Center, University of Pittsburgh, Pittsburgh, PA, 15260, USA

    Donna M. Huryn & Peter Wipf

  4. Department of Otolaryngology-Head and Neck Surgery, University of California, San Francisco, San Francisco, CA, 94118, USA

    Jennifer R. Grandis

  5. Clinical and Translational Science Institute, University of California, San Francisco, Box 0558, 550 16th Street, San Francisco, CA, 94143, USA

    Jennifer R. Grandis

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  1. Malabika Sen
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  2. Paul A. Johnston
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  3. Netanya I. Pollock
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  4. Kara DeGrave
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Correspondence to Jennifer R. Grandis.

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Funding sources

This project has been funded in part with Federal Funds from the National Cancer Institute, National Institutes of Health, under Contract No. HSN261200800001E. The content of this publication does not necessarily reflect the views or policies of the Department of Health and Human Services, nor does mention of trade names, commercial products, or organizations imply endorsement by the US Government. This study was supported by NExT-CBC Project ID #1015, S08-221 Task Order 6 “STAT3 Pathway Inhibitor HCS” (Grandis, PI), NCI Chemical Biology Consortium, Pittsburgh Specialized Application Center (PSAC) (Lazo and Johnston co-PIs), and University of Pittsburgh Chemical Diversity Center (Huryn, PI). The project was also supported in part by funds from the American Cancer Society (Grandis) and a Head and Neck Spore P50 award (Grandis, CA097190).

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Sen, M., Johnston, P.A., Pollock, N.I. et al. Mechanism of action of selective inhibitors of IL-6 induced STAT3 pathway in head and neck cancer cell lines. J Chem Biol 10, 129–141 (2017). https://doi.org/10.1007/s12154-017-0169-9

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