Issa Issa Ismail, Due Hanne, Brøndum Rasmus Froberg, Veeravakaran Vidthdyan, Haraldsdóttir Hulda, Sylvester Cathrine, Brogaard Asta, Dhanjal Soniya, Schmierer Bernhard, Dybkær Karen
Department of Hematology, Clinical Cancer Research Center, Aalborg University Hospital, 9000 Aalborg, Denmark.
Department of Clinical Medicine, Aalborg University, 9000 Aalborg, Denmark.
Cancers (Basel). 2024 Jul 2;16(13):2437. doi: 10.3390/cancers16132437.
The recurrence of diffuse large B-cell lymphoma (DLBCL) has been observed in 40% of cases. The standard of care for refractory/relapsed DLBCL (RR-DLBCL) is platinum-based treatment prior to autologous stem cell transplantation; however, the prognosis for RR-DLBCL patients remains poor. Thus, to identify genes affecting the cisplatin response in DLBCL, cisplatin-based whole-genome CRISPR-Cas9 knockout screens were performed in this study. We discovered DNA damage response (DDR) pathways as enriched among identified sensitizing CRISPR-mediated gene knockouts. In line, the knockout of the nucleotide excision repair genes and sensitized DLBCL cells to platinum drugs irrespective of proliferation rate, thus documenting DDR as essential for cisplatin sensitivity in DLBCL. Functional analysis revealed that the loss of and increased DNA damage levels and altered cell cycle distribution. Interestingly, we also identified which is involved in B-cell receptor signaling, to affect cisplatin response. The knockout of increased cisplatin sensitivity in DLBCL cells, and combinatory drug screens revealed a synergistic effect of the BTK inhibitor, ibrutinib, with platinum drugs at low concentrations. Applying local and external DLBCL cohorts, we addressed the clinical relevance of the genes identified in the CRISPR screens. was among the most frequently mutated genes with a frequency of 3-5%, and and were also mutated, albeit at lower frequencies. Furthermore, 27-54% of diagnostic DLBCL samples had mutations in pathways that can sensitize cells to cisplatin. In conclusion, this study shows that and , in addition to , are essential for the response to platinum-based drugs in DLBCL.
弥漫性大B细胞淋巴瘤（DLBCL）的复发在40%的病例中被观察到。难治性/复发性DLBCL（RR-DLBCL）的标准治疗是在自体干细胞移植前进行铂类治疗；然而，RR-DLBCL患者的预后仍然很差。因此，为了鉴定影响DLBCL中顺铂反应的基因，本研究进行了基于顺铂的全基因组CRISPR-Cas9敲除筛选。我们发现DNA损伤反应（DDR）通路在已鉴定的CRISPR介导的致敏基因敲除中富集。同样，核苷酸切除修复基因和的敲除使DLBCL细胞对铂类药物敏感，而与增殖率无关，从而证明DDR对DLBCL中顺铂敏感性至关重要。功能分析表明，和的缺失增加了DNA损伤水平并改变了细胞周期分布。有趣的是，我们还鉴定出参与B细胞受体信号传导的影响顺铂反应。的敲除增加了DLBCL细胞对顺铂的敏感性，联合药物筛选显示BTK抑制剂伊布替尼与低浓度铂类药物具有协同作用。应用局部和外部DLBCL队列，我们探讨了CRISPR筛选中鉴定的基因的临床相关性。是最常发生突变的基因之一，频率为3-5%，和也发生了突变，尽管频率较低。此外，27-54%的诊断性DLBCL样本在可使细胞对顺铂敏感的通路中发生了突变。总之，本研究表明，除了外，和对DLBCL中基于铂类药物的反应也至关重要。
