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Specific peptide interference reveals BCL6 transcriptional and oncogenic mechanisms in B-cell lymphoma cells

Abstract

The BTB/POZ transcriptional repressor and candidate oncogene BCL6 is frequently misregulated in B-cell lymphomas. The interface through which the BCL6 BTB domain mediates recruitment of the SMRT, NCoR and BCoR corepressors was recently identified. To determine the contribution of this interface to BCL6 transcriptional and biological properties, we generated a peptide that specifically binds BCL6 and blocks corepressor recruitment in vivo. This inhibitor disrupts BCL6-mediated repression and establishment of silenced chromatin, reactivates natural BCL6 target genes, and abrogates BCL6 biological function in B cells. In BCL6-positive lymphoma cells, peptide blockade caused apoptosis and cell cycle arrest. BTB domain peptide inhibitors may constitute a novel therapeutic agent for B-cell lymphomas.

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Figure 1: BBD motif peptides penetrate cells and bind to BCL6.
Figure 2: BBD motif peptides block BCL6 recruitment of SMRT.
Figure 3: Lateral groove corepressor blockade is specific for BCL6 and is required for transcriptional repression of natural and endogenous target genes.
Figure 4: Lateral groove blockade alters BCL6 target promoter repressosome composition and chromatin structure.
Figure 5: In vivo BCL6 lateral groove blockade causes abrogation of germinal center formation.
Figure 6: BCL6 lateral groove blockade causes apoptosis and cell cycle arrest of BCL6 dependent human B-cell lymphoma cells.

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Acknowledgements

This work was supported by US National Institutes of Health grants CA99982 (AMM) and CA59936 (JDL), the American Society of Hematology (AMM), Kimmel Foundation (AMM), the Chemotherapy Foundation (AMM and JDL), the National Cancer Institute-Canada (GGP) and the Burroughs Welcome Clinical Scientist Award (JDL). We thank T. Graf, B. Birshtein, T. Evans and R. Stanley for reading the manuscript, S. Dowdy for his advice on pTAT fusion proteins. We thank the Albert Einstein Analytical Imaging, Histopathology, Sequencing and FACS core facilities and especially M. Scharff and the members of his lab for technical support and S. Buhl at the Albert Einstein Hybridoma Core Facility for help in performing immunoassays.

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Correspondence to Ari Melnick.

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Supplementary information

Supplementary Fig. 1

In vitro and in vivo activities of wild-type and mutant peptide. (PDF 158 kb)

Supplementary Fig. 2

BCL6 lateral groove blockade does not induce inflammatory disease in mice. (PDF 183 kb)

Supplementary Methods (PDF 24 kb)

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Polo, J., Dell'Oso, T., Ranuncolo, S. et al. Specific peptide interference reveals BCL6 transcriptional and oncogenic mechanisms in B-cell lymphoma cells. Nat Med 10, 1329–1335 (2004). https://doi.org/10.1038/nm1134

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