Synergistic disruption of BTK and BCL-2 causes apoptosis while inducing ferroptosis in double-hit lymphoma

• Published in: European journal of pharmacology 2023-03, Vol.943, p.175526-175526, Article 175526
• Main Authors: Setiawan, Syahru Agung, Liu, Winston Zhenhao, Weng, Pei‐Wei, Lee, Chia-Hwa, Yadav, Vijesh Kumar, Hardianti, Mardiah Suci, Yeh, Chi-Tai, Chao, Tsu-Yi
• Format: Article
• Language: English
• Subjects: Apoptosis; BCL-2; BTK; Double-hit lymphoma; Ferroptosis; Humans; Lymphoma, Large B-Cell, Diffuse - drug therapy; Lymphoma, Large B-Cell, Diffuse - pathology; Proto-Oncogene Proteins c-bcl-2 - metabolism
• ISSN: 0014-2999; 1879-0712
• DOI: 10.1016/j.ejphar.2023.175526

Double-hit lymphoma (DHL) is an aggressive subset of Diffuse Large B-cell Lymphoma (DLBCL) with poor outcomes and without satisfying treatment options. BTK inhibitor monotherapy is ineffective to suppress aggressive lymphoma. Hence, combination with other potential agents is warranted. Here, we demonstrated the second generation of BTK inhibitor, zanubrutinib, and a BCL-2 inhibitor, navitoclax, worked in synergistic manner to suppress DHL. Comprehensive in silico approach by interrogating single-cell to bulk-level profiling was employed along with in vitro and in vivo validation in DHL cell lines. Ablation of BTK enhanced sensitivity to navitoclax and suppressed proliferation of DHL cells. Combination of second generation of BTK inhibitor with navitoclax synergistically suppressed DLBCL cells with higher synergy score in DHL subset. The drug combination triggered apoptosis and ferroptosis, with the latter being characterized by reactive oxygen species (ROS) accumulation, extensive lipid peroxidation, and depletion of reduced glutathione. Moreover, ablation of BTK sensitized DHL cells to ferroptosis. Mechanistically, disruption of BTK and BCL-2 triggered ferroptosis by downregulating NRF2 and HMOX1, while deactivating GPX4. Combination of zanubrutinib and navitoclax effectively suppressed tumor growth in vivo. Our data suggest that zanubrutinib and navitoclax synergistically suppressed DHL by inducing apoptosis and ferroptosis. [Display omitted] •Bruton's Tyrosine Kinase (BTK) maintained survival of Double-Hit Lymphoma (DHL).•Ablation of BTK Increased Sensitivity to BCL-2 Inhibition.•Disruption of BTK Enhanced Ferroptosis Sensitivity of DHL.•Inhibitor of BTK and BCL-2 Synergistically Suppressed DHL.•Zanubrutinib and Navitoclax Triggers Apoptosis and Ferroptosis in DHL.