Croatian Institute for Brain Research, University of Zagreb School of Medicine, 10 000, Zagreb, Croatia.
Department of Physiology, University of Zagreb School of Medicine, Zagreb, Croatia.
BMC Cancer. 2020 Nov 11;20(1):1090. doi: 10.1186/s12885-020-07533-6.
All-trans retinoic acid (ATRA)-based treatment of acute promyelocytic leukemia (APL) is the most successful pharmacological treatment of acute myeloid leukemia (AML). Recent development of inhibitors of mutated isocitrate dehydrogenase and dihydroorotate dehydrogenase (DHODH) has revived interest in differentiation therapy of non-APL AML. Our previous studies demonstrated that 5-aminoimidazole-4-carboxamide ribonucleoside (AICAr) induced differentiation of monocytic cell lines by activating the ATR/Chk1 via pyrimidine depletion. In the present study, the effects of AICAr on the viability and differentiation of primary AML blasts isolated from bone marrow of patients with non-APL AML were tested and compared with the effects of DHODH inhibitor brequinar and ATRA.
Bone marrow samples were obtained from 35 patients and leukemia blasts were cultured ex vivo. The cell viability was assessed by MTT assay and AML cell differentiation was determined by flow cytometry and morphological analyses. RNA sequencing and partial data analysis were conducted using ClusterProfiler package. Statistical analysis was performed using GraphPad Prism 6.0.
AICAr is capable of triggering differentiation in samples of bone marrow blasts cultured ex vivo that were resistant to ATRA. AICAr-induced differentiation correlates with proliferation and sensitivity to DHODH inhibition. RNA-seq data obtained in primary AML blasts confirmed that AICAr treatment induced downregulation of pyrimidine metabolism pathways together with an upregulation of gene set involved in hematopoietic cell lineage.
AICAr induces differentiation in a subset of primary non-APL AML blasts, and these effects correlate with sensitivity to a well-known, potent DHODH inhibitor.
全反式维甲酸（ATRA）为基础的治疗急性早幼粒细胞白血病（APL）是急性髓系白血病（AML）最成功的药物治疗方法。最近，异柠檬酸脱氢酶和二氢乳清酸脱氢酶（DHODH）抑制剂的发展重新激发了对非 APL AML 分化治疗的兴趣。我们之前的研究表明，5-氨基咪唑-4-甲酰胺核苷（AICAr）通过嘧啶耗竭激活 ATR/Chk1 诱导单核细胞系分化。在本研究中，检测了 AICAr 对非 APL AML 患者骨髓中分离的原代 AML 白血病细胞活力和分化的影响，并与 DHODH 抑制剂布雷奎纳和 ATRA 的影响进行了比较。
从 35 名患者中获得骨髓样本，并进行体外培养。通过 MTT 测定评估细胞活力，通过流式细胞术和形态学分析确定 AML 细胞分化。使用 ClusterProfiler 包进行 RNA 测序和部分数据分析。使用 GraphPad Prism 6.0 进行统计分析。
AICAr 能够触发体外培养的对 ATRA 耐药的骨髓白血病细胞分化。AICAr 诱导的分化与增殖和对 DHODH 抑制的敏感性相关。在原代 AML 白血病细胞中获得的 RNA-seq 数据证实，AICAr 处理诱导嘧啶代谢途径下调，同时上调与造血细胞谱系相关的基因集。
AICAr 诱导一部分原代非 APL AML 白血病细胞分化，这些作用与对一种已知的、有效的 DHODH 抑制剂的敏感性相关。
