Combination of Enzastaurin and Ibrutinib synergistically induces anti-tumor effects in diffuse large B cell lymphoma
Combination of Enzastaurin and Ibrutinib synergistically induces anti-tumor effects in diffuse large B cell lymphoma
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Enzastaurin 和 Ibrutinib 联合使用可协同诱导弥漫性大 B 细胞淋巴瘤的抗肿瘤作用
DOI：
10.1186/s13046-019-1076-4
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发表时间：
2019-02-18
影响因子：
11.3
通讯作者：
Zhu, Jun
中科院分区：
文献类型：
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作者：
He, Yizi;Li, Jiao;Zhu, Jun
背景弥漫性大B细胞淋巴瘤（DLBCL）是最常见的淋巴瘤。虽然超过一半的患者可以实现持久缓解，但DLBCL仍然是一个重大的临床挑战，约30%的患者无法治愈。BCR相关激酶（SYK，BTK和PI 3 K）抑制剂已显示出令人鼓舞的临床前和临床效果，如许多研究人员所报告的。早期研究表明蛋白激酶C-β（PKCβ）抑制剂可改变布鲁顿酪氨酸激酶（BTK）的磷酸化水平，从而增强BTK信号转导。在此，我们首次研究PKCβ抑制剂恩扎鲁因和BTK抑制剂伊曲替尼联合应用是否对DLBCL具有协同抗肿瘤作用。方法采用Cell Titer-Glo发光细胞活力测定法分析体外细胞增殖。流式细胞术检测细胞凋亡和细胞周期阻滞。Western Blotting分析检测相关信号通路中必需的调控酶。进行RNA-seq以评价低剂量恩扎鲁肽和伊曲替尼联合治疗带来的全转录组变化。结果恩扎鲁明与伊鲁替尼联合应用对DLBCL细胞的存活和增殖具有持久的协同作用，包括抑制细胞增殖、促进细胞凋亡、诱导G1期阻滞、阻止细胞侵袭和迁移、下调下游信号的激活等。更重要的是，全转录组变化结果表明，与恩扎鲁肽和伊曲替尼单独治疗相比，联合治疗协同调节全转录组表达。低剂量恩扎鲁肽与伊曲替尼联合应用可有效下调BCR、NF-κB、JAK和MAPK相关信号通路。此外，mRNA表达分析进一步表明，共同处理显着降低NOTCH 1的mRNA水平。抑制DLBCL细胞增殖的联合作用可能是通过抑制NOTCH 1的表达实现的。最后，联合治疗的抗肿瘤活性也被证明在vivo.ConclusionsCombination恩扎鲁肽和伊曲替尼在DLBCL中具有协同抗肿瘤作用，独立于分子亚型。这些结果为DLBCL的治疗提供了良好的基础，同时抑制BTK和PKCβ可能成为DLBCL治疗的新策略。
BackgroundDiffuse large B cell lymphoma (DLBCL) is the most common form of lymphoma. Although durable remissions can be achieved in more than half of these patients, DLBCL remains a significant clinical challenge, with approximately 30% of patients not being cured. BCR-associated kinases (SYK, BTK, and PI3K) inhibitors have exhibited encouraging pre-clinical and clinical effects, as reported by many researchers. Early studies demonstrated that protein kinase C-β (PKCβ) inhibitors alter phosphorylation level the Bruton’s tyrosine kinase (BTK), which leads to enhanced BTK signaling. Here, for the first time, we investigate whether the combination of PKCβ inhibitor enzastaurin and BTK inhibitor ibrutinib has synergistic anti-tumor effects in DLBCL.MethodsIn vitro cell proliferation was analyzed using Cell Titer-Glo Luminescent Cell Viability Assay. Induction of apoptosis and cell cycle arrest were measured by flow cytometry. Western Blotting analysis was used to detect the essential regulatory enzymes in related signaling pathways. RNA-seq was conducted to evaluate the whole transcriptome changes brought by co-treatment with low doses of enzastaurin and ibrutinib. The synergistic anti-tumor effects of enzastaurin and ibrutinib were also evaluated in vivo.ResultsCombination of enzastaurin and ibrutinib produced a lasting synergistic effect on the survival and proliferation of DLBCL cells, including reduction of proliferation, promoting apoptosis, inducting G1 phase arrest, preventing cell invasion and migration, and down-regulating activation of downstream signaling. More importantly, whole-transcriptome changes results showed that combination therapy worked synergistically to regulate whole-transcriptome expression compared with enzastaurin and ibrutinib alone. Co-treatment with low doses of enzastaurin and ibrutinib could effectively downregulate BCR, NF-κB, JAK and MAPK related signaling pathway. Furthermore, the mRNA expression analysis further indicated that co-treatment significantly decreased the mRNA levels of NOTCH1. The combination effect in inhibiting proliferation of DLBCL cells probably was realized through suppression of NOTCH1 expression. Finally, the anti-tumor activity of co-treatment also was demonstrated in vivo.ConclusionsCombination of enzastaurin and ibrutinib had synergistic anti-tumor effects in DLBCL, independent of molecular subtype. These results provided a sound foundation for an attractive therapeutic treatment, and the simultaneous suppression of BTK and PKCβ might be a new treatment strategy for DLBCL.