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Supplementary Figures S1-S6 and Tables S1-S2 from SF2523: Dual PI3K/BRD4 Inhibitor Blocks Tumor Immunosuppression and Promotes Adaptive Immune Responses in Cancer

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posted on 2023-04-03, 15:21 authored by Shweta Joshi, Alok R. Singh, Kevin X. Liu, Timothy V. Pham, Muamera Zulcic, Dylan Skola, Hyun Bae Chun, Christopher K. Glass, Guillermo A. Morales, Joseph R. Garlich, Donald L. Durden

Supplementary Fig. S1 shows JQ1 blocks IL4 induced CD206 expression. Supplementary Fig. S2 shows Differentially expressed genes in LPS-stimulated or IL4 stimulated BMDMs. Supplementary Fig. S3 shows Effect of JQ1 on LLC tumor growth. Supplementary Fig. S4 shows SF1126 blocks tumor growth and immunosuppressive macrophage polarization. Supplementary Fig. S5 shows JQ1 and SF2523 reduce tumor growth due to its effect on macrophages. Supplementary Fig. S6 shows SF2523 inhibits B16 tumor metastasis. Supplementary Table S1 shows Primers used for ChIP analysis. Supplementary Table S2 shows Contingency tables representing the effects on the number of shared over or under expressed genes of different exposures.

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ARTICLE ABSTRACT

Macrophages (MΘs) are key immune infiltrates in solid tumors and serve as major drivers behind tumor growth, immune suppression, and inhibition of adaptive immune responses in the tumor microenvironment (TME). Bromodomain and extraterminal (BET) protein, BRD4, which binds to acetylated lysine on histone tails, has recently been reported to promote gene transcription of proinflammatory cytokines but has rarely been explored for its role in IL4-driven MΘ transcriptional programming and MΘ-mediated immunosuppression in the TME. Herein, we report that BET bromodomain inhibitor, JQ1, blocks association of BRD4 with promoters of arginase and other IL4-driven MΘ genes, which promote immunosuppression in TME. Pharmacologic inhibition of BRD4 using JQ1 and/or PI3K using dual PI3K/BRD4 inhibitor SF2523 (previously reported by our group as a potent inhibitor to block tumor growth and metastasis in various cancer models) suppresses tumor growth in syngeneic and spontaneous murine cancer models; reduces infiltration of myeloid-derived suppressor cells; blocks polarization of immunosuppressive MΘs; restores CD8+ T-cell activity; and stimulates antitumor immune responses. Finally, our results suggest that BRD4 regulates the immunosuppressive myeloid TME, and BET inhibitors and dual PI3K/BRD4 inhibitors are therapeutic strategies for cancers driven by the MΘ-dependent immunosuppressive TME.

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