A FOXO1-dependent transcription network is a targetable vulnerability of mantle cell lymphomas
Ja-Young Jang, … , Hongwu Zheng, Jihye Paik
Ja-Young Jang, … , Hongwu Zheng, Jihye Paik
Published October 25, 2022
Citation Information: J Clin Invest. 2022;132(24):e160767. https://doi.org/10.1172/JCI160767.
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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 3

FOXO1 acts upstream of the MCL lineage regulatory TF hierarchy.

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FOXO1 acts upstream of the MCL lineage regulatory TF hierarchy. (A) Immu...
(A) Immunoblot analysis of CCMCL1 cells transduced with EBF1-V5 (left), 3× FLAG–tagged IRF4 (middle), or 3× FLAG–tagged PAX5 (right). (B) Competition-based proliferation assays of sgRNAs against FOXO1 in Cas9-transduced CCMCL1 cells expressing mock control, EBF1-V5, or 3× FLAG–tagged IRF4 or PAX5. (C) Competition-based proliferation assays of sgRNAs against EBF1, IRF4, or PAX5 in control and FOXO1r#1-transduced (resistant for sgRNA#1 of FOXO1) CCMCL1 cells. In B and C, data represent mean ± SEM (n = 3). Results are representative of 3 independent experiments. Statistical analysis was performed using 2-tailed unpaired Student’s t test. The day 13 values of each cell line responding to the same sgRNA were respectively compared. **P < 0.001 ***P < 0.0005, ****P < 0.0001. (D) Immunoblot analysis of CCMCL1 (left) or JEKO1 (right) cells depleted of FOXO1. Lysates were prepared at day 3 after infection of indicated sgRNAs targeting FOXO1. (E–H) Immunoblot and RT-qPCR analysis of EBF1, IRF4, or PAX5 induction in FOXO1-transduced HEL (E and F) or THP1 cells (G and H). Cell lysates and total RNA were prepared at indicated time points after infection of FOXO1-encoding lentivirus. In F and H, 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. *P < 0.05, **P < 0.005, ***P < 0.0005, ****P < 0.0001.