Abstract
Purpose
Double-hit lymphoma (DHL) is a rare and aggressive mature B-cell malignancy with concurrent MYC and BCL2 rearrangements. When DHL becomes relapsed or refractory, it becomes resistant to the majority of therapeutic approaches and has subpar clinical results. Therefore, innovative therapeutics for this particular patient population are urgently needed.
Methods
Orelabrutinib, a new oral BTK inhibitor, combined with the Bcl-2 inhibitor venetoclax, was used to confirm the antitumor effect of DHL. Cell counting kit-8 and Annexin V-FITC/PI assays were used to examine the interaction of this combined regimen on DHL cell lines and primary lymphoma cells. RNA sequencing, EdU incorporation assay, mitochondrial membrane potential assay, and western blotting were employed to explore the molecule mechanism for the cytotoxicity of orelabrutinib with or without venetoclax against DHL cell lines.
Results
In this study, orelabrutinib combined with venetoclax synergistically induced DHL cell death, as evidenced by the inhibition of cell proliferation, the induct of cell cycle arrest, and the promotion of cell apoptosis via the mitochondrial pathway. Orelabrutinib treatment alters genome-wide gene expression in DHL cells. The combined regimen decreases the expression of BTK and Mcl-1, potentially interfering with the activity and crosstalk of PI3K/AKT signaling and p38/MAPK signaling. In addition, the combination of orelabrutinib and venetoclax shows cytotoxic activity in primary B-lymphoma cells.
Conclusion
In summary, these findings reveal a novel therapy targeting BCR signaling and the Bcl-2 family for DHL patients with a poor prognosis.
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Data availability
The datasets generated during the current study are available from the corresponding author on reasonable request.
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Funding
This work was supported by the National Natural Science Foundation of China (82170180, 82100204), the Natural Science Foundation of Fujian Province (2020J011246, 2021J011359), the Xiamen Municipal Bureau of Science and Technology (3502Z20209003) and Dongguan Science and Technology Bureau (No.202050715001214).
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Conceptualization, JZ and BX; methodology, GCP, MYZ, and JWY; software, GCP and JST; validation, MYZ, YLJ, YRJ, JZ, and BX; formal analysis, GCP, YLJ, YFT, HZ, and DMQ; resources, XXY, LL, ZFL, ZJL, YRJ, and BX; data curation, GCP, MYZ, JZ, and BX; writing—original draft preparation, GCP and MYZ; writing—review and editing, MYZ and JWY; supervision, JZ and BX; funding acquisition, JZ, YRJ, and BX; all the authors have read and approved the final manuscript.
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This study was performed in line with the principles of the Declaration of Helsinki. Approval was granted by the First Affiliated Hospital of Xiamen University Ethics Review Board. Informed consent was obtained from the patients before the collection of lymph node samples.
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432_2022_4473_MOESM2_ESM.tif
Supplementary file2 Supplementary Fig. 2 Flow cytometry detection of orelabrutinib-induced apoptosis in DHL cells. A–F The apoptosis statistics for TMD8, LR, MCA, and WILL-2 were calculated using Prism 8 (TIF 3341 KB)
432_2022_4473_MOESM3_ESM.tif
Supplementary file3 Supplementary Fig. 3 GSEA of genes affected by orelabrutinib treatment. Alterations in A p53 signaling pathways and B hematopoietic cell lineages following treatment with TMD8 and 10 μM orelabrutinib. C Western blot of PI3K/AKT, p38/MAPK, and NF-κB signaling proteins in WILL-2 cells exposed to 50 μM orelabrutinib, 300 nM ABT-199 or both (TIF 5014 KB)
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Pan, G., Zhong, M., Yao, J. et al. Orelabrutinib and venetoclax synergistically induce cell death in double-hit lymphoma by interfering with the crosstalk between the PI3K/AKT and p38/MAPK signaling. J Cancer Res Clin Oncol 149, 5513–5529 (2023). https://doi.org/10.1007/s00432-022-04473-5
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DOI: https://doi.org/10.1007/s00432-022-04473-5