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Interfering with bromodomain epigenome readers as therapeutic option in mucoepidermoid carcinoma

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Abstract

Purpose

Emerging evidence indicates that bromodomains comprise a conserved class of epigenome readers involved in cancer development and inflammation. Bromodomains are associated with epigenetic modifications of gene transcription through interactions with lysine residues of histone tails. Particularly, the bromodomain and extra-terminal domain (BET) family member BRD4 has been found to be involved in the control over oncogenes, including c-MYC, and in the maintenance of downstream inflammatory processes. The objective of this study was to evaluate the effect of pharmacologically displacing BRD4 in mucoepidermoid carcinoma (MEC) cells.

Methods

We assessed the presence of BRD4 levels in a panel of human MEC tissue samples in conjunction with histological grading and clinical information. In vitro studies were carried out using human MEC-derived cell lines. The BET inhibitor iBET762 was administered to MEC cells to assess the impact of disrupted BRD4 signaling on colony forming capacities and cell cycle status. The activation of cellular senescence induced by iBET762 was determined by immunohistochemical staining for p16ink4. Flow cytometry was used to identify populations of cancer stem cells in MEC-derived cell lines.

Results

We found that primary human MECs and MEC-derived cell lines are endowed with high BRD4 expression levels compared to those in normal salivary glands. We also found that, by displacing BRD4 from chromatin using the BET inhibitor iBET762, MEC cells lose their colony forming capacities and undergo G1 cell cycle arrest and senescence. Finally, we found that targeted displacement of BRD4 from chromatin results in depletion of cancer stem cells from the overall MEC cell populations.

Conclusions

Our findings indicate that bromodomain-mediated gene regulation constitutes an epigenetic mechanism that is deregulated in MEC cells and that the use of BET inhibitors may serve as a feasible therapeutic strategy to manage MECs.

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Acknowledgments

This work was conducted during a visiting scholar period at the University of Michigan, sponsored by the Capes Foundation within the Ministry of Education, Brazil (grant n. BEX / 88881.132606/2016-01 PDSE). This grant was funded by a University of Michigan School of Dentistry faculty grant, and a Cancer Center Support Grant (P30 CA046592). The authors declare no potential conflicts of interest with respect to authorships and/or publication of this article.

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Authors and Affiliations

  1. Laboratory of Epithelial Biology, Department of Periodontics and Oral Medicine, University of Michigan School of Dentistry, 1011 N University Ave, Room 2029C, Ann Arbor, MI, 48109-1078, USA

    Renata L. Markman, Liana P. Webber, Carlos H. V. Nascimento Filho, Leonardo A. Reis, Cristiane H. Squarize & Rogerio M. Castilho

  2. Department of Oral Diagnosis, Piracicaba Dental School, State University of Campinas-UNICAMP, Piracicaba, Brazil

    Renata L. Markman, Leonardo A. Reis, Pablo A. Vargas & Marcio A. Lopes

  3. Department of Oral Pathology, School of Dentistry, Federal University of Rio Grande do Sul-UFRGS, Porto Alegre, RS, 90035-003, Brazil

    Liana P. Webber, Virgilio Zanella & Manoela D. Martins

  4. Genetics and Molecular Biology Research Unit, Sao Jose do Rio Preto Medical School-FAMERP, Sao Jose do Rio Preto, Brazil

    Carlos H. V. Nascimento Filho

  5. Head and Neck Department, Santa Rita Hospital - Santa Casa de Misericordia de Porto Alegre, Porto Alegre, RS, Brazil

    Virgilio Zanella

  6. Comprehensive Cancer Center, University of Michigan Ann Arbor, Ann Arbor, MI, 48109, USA

    Cristiane H. Squarize & Rogerio M. Castilho

Authors
  1. Renata L. Markman
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  2. Liana P. Webber
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  3. Carlos H. V. Nascimento Filho
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  9. Cristiane H. Squarize
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  10. Rogerio M. Castilho
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Contributions

RLM performed most of the cell culture-based assays, IHC, IF, and participated in the organization of the figures. CHVNF performed the flow cytometry assays, LAR helped with the IHC and IF assays, and LPW helped with the immunofluorescence assays and its quantifications. VZ and MDM contributed to the collection of MEC tissue samples and their clinical data and helped with their re-evaluation. PAV, MAL, CHS and RMC contributed to the conception, design, data organization and writing of the manuscript. All authors gave their final approval and agreed to be accountable for all aspects of the work.

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Correspondence to Rogerio M. Castilho.

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Markman, R.L., Webber, L.P., Nascimento Filho, C.H.V. et al. Interfering with bromodomain epigenome readers as therapeutic option in mucoepidermoid carcinoma. Cell Oncol. 42, 143–155 (2019). https://doi.org/10.1007/s13402-018-0416-2

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