A FOXO1-dependent transcription network is a targetable vulnerability of mantle cell lymphomas
Ja-Young Jang, Inah Hwang, Heng Pan, Jun Yao, Lapo Alinari, Eddie Imada, Claudio Zanettini, Michael J. Kluk, Yizhe Wang, Yunkyoung Lee, Hua V. Lin, Xiangao Huang, Maurizio Di Liberto, Zhengming Chen, Karla V. Ballman, Lewis C. Cantley, Luigi Marchionni, Giorgio Inghirami, Olivier Elemento, Robert A. Baiocchi, Selina Chen-Kiang, Sandro Belvedere, Hongwu Zheng, Jihye Paik
Ja-Young Jang, Inah Hwang, Heng Pan, Jun Yao, Lapo Alinari, Eddie Imada, Claudio Zanettini, Michael J. Kluk, Yizhe Wang, Yunkyoung Lee, Hua V. Lin, Xiangao Huang, Maurizio Di Liberto, Zhengming Chen, Karla V. Ballman, Lewis C. Cantley, Luigi Marchionni, Giorgio Inghirami, Olivier Elemento, Robert A. Baiocchi, Selina Chen-Kiang, Sandro Belvedere, Hongwu Zheng, Jihye Paik
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Research Article Oncology

A FOXO1-dependent transcription network is a targetable vulnerability of mantle cell lymphomas

Abstract

Targeting lineage-defined transcriptional dependencies has emerged as an effective therapeutic strategy in cancer treatment. Through screening for molecular vulnerabilities of mantle cell lymphoma (MCL), we identified a set of transcription factors (TFs) including FOXO1, EBF1, PAX5, and IRF4 that are essential for MCL propagation. Integrated chromatin immunoprecipitation and sequencing (ChIP-Seq) with transcriptional network reconstruction analysis revealed FOXO1 as a master regulator that acts upstream in the regulatory TF hierarchy. FOXO1 is both necessary and sufficient to drive MCL lineage commitment through supporting the lineage-specific transcription programs. We further show that FOXO1, but not its close paralog FOXO3, can reprogram myeloid leukemia cells and induce B-lineage gene expression. Finally, we demonstrate that cpd10, a small molecule identified from an enriched FOXO1 inhibitor library, induces a robust cytotoxic response in MCL cells in vitro and suppresses MCL progression in vivo. Our findings establish FOXO1 inhibition as a therapeutic strategy targeting lineage-driven transcriptional addiction in MCL.

Authors

Ja-Young Jang, Inah Hwang, Heng Pan, Jun Yao, Lapo Alinari, Eddie Imada, Claudio Zanettini, Michael J. Kluk, Yizhe Wang, Yunkyoung Lee, Hua V. Lin, Xiangao Huang, Maurizio Di Liberto, Zhengming Chen, Karla V. Ballman, Lewis C. Cantley, Luigi Marchionni, Giorgio Inghirami, Olivier Elemento, Robert A. Baiocchi, Selina Chen-Kiang, Sandro Belvedere, Hongwu Zheng, Jihye Paik

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Figure 6

Cpd10 is a FOXO1-specific inhibitor that suppresses MCL growth in vitro.

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Cpd10 is a FOXO1-specific inhibitor that suppresses MCL growth in vitro....
(A) Ranking of cell death induction activity of 144 FOXO1 small-molecule inhibitors. Cell death was measured by TO-PRO-3 cell death assay (Thermo Fisher Scientific) after 6-day treatment of compounds (4 μM) in 4 indicated cell lines. Red arrows point to the selected inhibitors for follow-up studies. (B–D) Dose-dependent effect of FOXO1 inhibitors on MCL and AML cell lines. Relative cell death (percent) was determined by TO-PRO-3 staining at day 6 under treatment of the indicated compounds. (E) Growth curve of CCMCL1, HEL, and THP1 cells under cpd10 (2 μM). (F) Percentage of annexin V–positive JEKO1, HEL, and THP1 cells at day 6 following treatment with cpd10 (2 μM). (G) Cell cycle distribution of cells treated with cpd10 (2 μM). (H) Primary MCL cells were cultured with cpd10 (2 μM) or vehicle. Data represent mean ± SEM (n = 4, MCL1, MCL4; n = 3, MCL2, MCL3). Results are representative of 2 independent experiments. (I and J) Immunoblot (I) and RT-qPCR assay (J) of CD79B, EBF1, IRF4, or PAX5 mRNA expression in 3 indicated control and cpd10-treated cell lines. Total RNAs were prepared after 48 hours of treatment. (K) Representative proximity ligation assay (PLA) image of MAVER1 cells demonstrating the inhibition of interaction between FOXO1 and p300 in response to 2 μM cpd10. Scale bars: 20 μm. (L) Quantitation of mean fluorescence intensity (MFI) of PLA signal per nuclei from MAVER1 or CCMCL1 cells. The number of nuclei scored is indicated. (B–F, J, and L) Data represent mean ± SEM (n = 3). Results are representative of 3 independent experiments. Statistical analysis was performed using 1-way ANOVA with Tukey’s multiple-comparison test in B–D and using 2-tailed unpaired Student’s t test in E, F, H, J, and L. *P < 0.05, **P < 0.001, ***P < 0.0005, ****P < 0.0001.