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Ezh2Y641F mutations co-operate with Stat3 to regulate MHC class I antigen processing and alter the tumor immune response in melanoma

Oncogene volume 41, pages 4983–4993 (2022)Cite this article

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Abstract

Enhancer of Zeste Homolog 2 (EZH2) is the catalytic component of the Polycomb Repressive Complex 2, a chromatin modifying complex, which mediates methylation of lysine 27 on histone 3 (H3K27me3), a repressive chromatin mark. Genetic alterations in EZH2 in melanoma include amplifications and activating point mutations at tyrosine 641 (Y641) whose underlying oncogenic mechanisms remain largely unknown. Here, we found that expression of Ezh2Y641F causes upregulation of a subset of interferon-regulated genes in melanoma cells. Upregulation of these genes was not a direct effect of changes in H3K27me3, but via a non-canonical interaction between Ezh2 and Signal Transducer and Activator of Transcription 3 (Stat3). Ezh2 and Stat3 together function as transcriptional activators to mediate gene activation of numerous genes, including MHC Class 1b antigen processing genes. Furthermore, expression of Stat3 is required to maintain an anti-tumor immune response in Ezh2Y641F melanomas and to prevent melanoma progression and recurrence.

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Fig. 1: EZH2Y641F melanomas exhibit an interferon-related gene expression profile independent of H3K27me3.
Fig. 2: Stat3 co-immunoprecipitates with Ezh2 in Ezh2Y641F mutant cells and is hypermethylated.
Fig. 3: Stat3 knockdown in melanoma cell lines partially restores IFN-related gene expression without affecting intrinsic cell growth.
Fig. 4: Changes in global chromatin binding by Ezh2 and Stat3 in Ezh2Y641F melanoma cells and its effect on gene expression.
Fig. 5: Direct regulation of MHC Class 1b H2-Q genes by Ezh2 and Stat3.
Fig. 6: Assessment of tumor growth and tumor-infiltrating immune cells in Ezh2WT vs Ezh2Y641F melanoma cells.
Fig. 7: Analysis of tumor growth over time of Ezh2WT vs Ezh2Y641F melanoma cells after Stat3 knockdown.

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Acknowledgements

We thank the Siteman Flow Cytometry facility, McDonnel Genome Institute/Genome Access Center for technical assistance, and the Department of Comparative Medicine for animal expertise. We also thank all members of the Souroullas lab for critical input on the manuscript.

Funding

This work was supported by the US National Cancer Institute K22-CA229612-01(GPS) and T32 CA113275-10 (SZ), the Cancer Research Foundation, Chicago IL, (GPS), The Harry J. Lloyd Charitable Trust (GPS).

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Authors and Affiliations

  1. Department of Medicine, Washington University School of Medicine in St. Louis, St. Louis, MO, 63110, USA

    Sarah M. Zimmerman, Samantha J. Nixon, Pei Yu Chen, Leela Raj, Sofia R. Smith, Rachel L. Paolini, Phyo Nay Lin & George P. Souroullas

  2. Division of Oncology, Molecular Oncology Section, Washington University School of Medicine in St. Louis, St. Louis, MO, 63110, USA

    Sarah M. Zimmerman, Samantha J. Nixon, Pei Yu Chen, Leela Raj, Sofia R. Smith, Rachel L. Paolini, Phyo Nay Lin & George P. Souroullas

  3. Siteman Comprehensive Cancer Center, Washington University School of Medicine in St. Louis, St. Louis, MO, 63110, USA

    George P. Souroullas

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Contributions

GPS and SMZ designed experiments and wrote the manuscript. GPS, SJN and SMZ performed experiments, analyzed, and interpreted the data. LR, PYC, RLP, SS and PNL performed experiments. GPS conceived of and supervised the study.

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Correspondence to George P. Souroullas.

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Zimmerman, S.M., Nixon, S.J., Chen, P.Y. et al. Ezh2Y641F mutations co-operate with Stat3 to regulate MHC class I antigen processing and alter the tumor immune response in melanoma. Oncogene 41, 4983–4993 (2022). https://doi.org/10.1038/s41388-022-02492-7

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