Department of Tumor Immunotherapy, Shenzhen Luohu People's Hospital, The Third Affiliated Hospital of Shenzhen University, Shenzhen, Guangdong, P.R. China.
Medical Laboratory of Shenzhen Luohu People's Hospital, Shenzhen, China.
Cancer Med. 2022 Sep;11(18):3479-3490. doi: 10.1002/cam4.4712. Epub 2022 Apr 29.
Significant progress has been made in the diagnosis and treatment of the drug-resistant and highly recurrent refractory T cell acute lymphoblastic leukemia (T-ALL). Primary tumor cell-derived induced pluripotent stem cells (iPSCs) have become very useful tumor models for cancer research including drug sensitivity tests. In the present study, we investigated the mechanism underlying drug resistance in T-ALL using the T-ALL-derived iPSCs (T-iPSCs) model. T-ALL cells were transformed using iPSC reprogramming factors (Sox-2, Klf4, Oct4, and Myc) via nonintegrating Sendai virus. T-iPSCs with the Notch1 mutation were then identified through genomic sequencing. Furthermore, T-iPSCs resistant to 80 μM LY411575, a γ-secretase and Notch signal inhibitor, were also established. We found a significant difference in the expression of drug resistance-related genes between the drug-resistant T-iPSCs and drug-sensitive groups. Among the 27 genes, six most differently expressed genes (DEGs) based on Log FC >5 were identified. Knockdown analyses using RNA interference (RNAi) revealed that MAEL is the most important gene associated with drug resistance in T-ALL cells. Also, MAEL knockdown downregulated expression of MRP and LRP in drug-resistant T-iPSCs. Interestingly, this phenomenon partially restored the sensitivity of the cells to LY411575. Furthermore, overexpression of the MAEL gene enhanced drug resistance against LY411575. Conclusively, MAEL promotes LY411575 resistance in T-ALL cells increasing the expression of MRP and LRP genes.
在耐药性和高度复发性难治性 T 细胞急性淋巴细胞白血病（T-ALL）的诊断和治疗方面已经取得了显著进展。原代肿瘤细胞衍生的诱导多能干细胞（iPSCs）已成为癌症研究非常有用的肿瘤模型，包括药物敏感性测试。在本研究中，我们使用 T-ALL 衍生的 iPSCs（T-iPSCs）模型研究了 T-ALL 耐药的机制。T-ALL 细胞通过非整合性仙台病毒使用 iPSC 重编程因子（Sox-2、Klf4、Oct4 和 Myc）进行转化。然后通过基因组测序鉴定出具有 Notch1 突变的 T-iPSCs。此外，还建立了对 80 μM LY411575（一种 γ-分泌酶和 Notch 信号抑制剂）耐药的 T-iPSCs。我们发现耐药性 T-iPSCs 和敏感性组之间药物耐药相关基因的表达存在显著差异。在 27 个基因中，根据 Log FC>5 确定了 6 个表达差异最大的基因（DEGs）。使用 RNA 干扰（RNAi）的敲低分析显示，MAEL 是与 T-ALL 细胞耐药性最相关的最重要基因。此外，MAEL 敲低降低了耐药性 T-iPSCs 中 MRP 和 LRP 的表达。有趣的是，这种现象部分恢复了细胞对 LY411575 的敏感性。此外，MAEL 基因的过表达增强了 T-ALL 细胞对 LY411575 的耐药性。总之，MAEL 通过增加 MRP 和 LRP 基因的表达促进了 T-ALL 细胞对 LY411575 的耐药性。
