Hyperphosphorylation of BCL-2 family proteins underlies functional resistance to venetoclax in lymphoid malignancies
Stephen Jun Fei Chong, Fen Zhu, Olga Dashevsky, Rin Mizuno, Jolin X.H. Lai, Liam Hackett, Christine E. Ryan, Mary C. Collins, J. Bryan Iorgulescu, Romain Guièze, Johany Penailillo, Ruben Carrasco, Yeonjoo C. Hwang, Denise P. Muñoz, Mehdi Bouhaddou, Yaw Chyn Lim, Catherine J. Wu, John N. Allan, Richard R. Furman, Boon Cher Goh, Shazib Pervaiz, Jean-Philippe Coppé, Constantine S. Mitsiades, Matthew S. Davids
Stephen Jun Fei Chong, Fen Zhu, Olga Dashevsky, Rin Mizuno, Jolin X.H. Lai, Liam Hackett, Christine E. Ryan, Mary C. Collins, J. Bryan Iorgulescu, Romain Guièze, Johany Penailillo, Ruben Carrasco, Yeonjoo C. Hwang, Denise P. Muñoz, Mehdi Bouhaddou, Yaw Chyn Lim, Catherine J. Wu, John N. Allan, Richard R. Furman, Boon Cher Goh, Shazib Pervaiz, Jean-Philippe Coppé, Constantine S. Mitsiades, Matthew S. Davids
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Research Article Hematology Oncology

Hyperphosphorylation of BCL-2 family proteins underlies functional resistance to venetoclax in lymphoid malignancies

Abstract

The B cell leukemia/lymphoma 2 (BCL-2) inhibitor venetoclax is effective in chronic lymphocytic leukemia (CLL); however, resistance may develop over time. Other lymphoid malignancies such as diffuse large B cell lymphoma (DLBCL) are frequently intrinsically resistant to venetoclax. Although genomic resistance mechanisms such as BCL2 mutations have been described, this probably only explains a subset of resistant cases. Using 2 complementary functional precision medicine techniques — BH3 profiling and high-throughput kinase activity mapping — we found that hyperphosphorylation of BCL-2 family proteins, including antiapoptotic myeloid leukemia 1 (MCL-1) and BCL-2 and proapoptotic BCL-2 agonist of cell death (BAD) and BCL-2 associated X, apoptosis regulator (BAX), underlies functional mechanisms of both intrinsic and acquired resistance to venetoclax in CLL and DLBCL. Additionally, we provide evidence that antiapoptotic BCL-2 family protein phosphorylation altered the apoptotic protein interactome, thereby changing the profile of functional dependence on these prosurvival proteins. Targeting BCL-2 family protein phosphorylation with phosphatase-activating drugs rewired these dependencies, thus restoring sensitivity to venetoclax in a panel of venetoclax-resistant lymphoid cell lines, a resistant mouse model, and in paired patient samples before venetoclax treatment and at the time of progression.

Authors

Stephen Jun Fei Chong, Fen Zhu, Olga Dashevsky, Rin Mizuno, Jolin X.H. Lai, Liam Hackett, Christine E. Ryan, Mary C. Collins, J. Bryan Iorgulescu, Romain Guièze, Johany Penailillo, Ruben Carrasco, Yeonjoo C. Hwang, Denise P. Muñoz, Mehdi Bouhaddou, Yaw Chyn Lim, Catherine J. Wu, John N. Allan, Richard R. Furman, Boon Cher Goh, Shazib Pervaiz, Jean-Philippe Coppé, Constantine S. Mitsiades, Matthew S. Davids

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Figure 5

Reduction in BCL-2 family protein phosphorylation rewires resistant cells to BCL-2 dependence.

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Reduction in BCL-2 family protein phosphorylation rewires resistant cell...
(A) Diagram demonstrating the DBP technique. Cells were treated with the drug of interest prior to BH3 profiling. DBP identifies whether a drug of interest changes the antiapoptotic dependence(ies) of cells. The diagram was generated using BioRender software. (B and C) DBP of Su-DHL4 (n = 5) and OCI-Ly1-R (n = 3) cells following treatment with FTY720 (10 μM) for 4 hours. (D) Δ Changes in the percentage of Cytc loss between FTY720 and DMSO treatments in the acquired-resistance and intrinsically resistant cell lines (n = 5 for Su-DHL4, n = 3 for all other cell lines). (E) Percentage of Cytc loss with ABT199/venetoclax (0.5 μM) following a 4-hour treatment with FTY720 in pcDNA3.1-, WT BCL-2–, p.S70A-, or p.S70E-transfected OCI-Ly1-R cells (n = 3). Šidák’s multiple-comparison test was used. (F) Percentage of Cytc loss with MS1 peptide (5 μM) following a 4-hour FTY720 treatment in pcDNA3.1-, WT MCL-1–, or p.T163A-transfected Su-DHL4 cells (n = 3). Dunnett’s multiple-comparison test was used. *P < 0.05, **P < 0.01, and ***P < 0.001.