Hyperphosphorylation of BCL-2 family proteins underlies functional resistance to venetoclax in lymphoid malignancies
Stephen Jun Fei Chong, … , Constantine S. Mitsiades, Matthew S. Davids
Stephen Jun Fei Chong, … , Constantine S. Mitsiades, Matthew S. Davids
Published September 26, 2023
Citation Information: J Clin Invest. 2023;133(22):e170169. https://doi.org/10.1172/JCI170169.
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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 2

Venetoclax-resistant malignant lymphoid cells display increased MCL-1 dependence and decreased BCL-2 dependence.

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Venetoclax-resistant malignant lymphoid cells display increased MCL-1 de...
(A) Diagram showing the baseline BH3-profiling technique, in which cells are required to be incubated with BH3 peptides/mimetic to induce Cytc release. Heatmap demonstrates the sensitivity/dependence of antiapoptotic BCL-2 family members toward their specific BH3 peptides/mimetic, as represented by the intensity of Cytc release in red. Higher Cytc release indicates a greater dependence on an antiapoptotic protein(s). The diagram was generated using BioRender software. (B) Baseline BH3 profiling of malignant lymphoid cell lines, depicted by a heatmap of Cytc loss intensity following individual BH3 peptides or ABT199/venetoclax incubation (n = 3). Higher Cytc release by MS1 indicates MCL-1 dependence, and lower Cytc release by BAD or venetoclax/ABT199 indicates lower BCL-2 dependence. (C) Delta (Δ) changes in the percentage of Cytc loss were calculated by deducting the percentage of Cytc loss of sensitive cell lines from that of resistant cell lines or the percentage of Cytc loss of vehicle-treated cells from that of drug-treated cells. (D) Δ Changes in the percentage of Cytc loss indicate an increase in MCL-1 dependency and a drop in BCL-2 dependency. A positive Δ change in the percentage indicates increased net Cytc loss, and a negative Δ change in the percentage indicates decreased net Cytc loss.