Identification and characterization of isocitrate dehydrogenase 1 (IDH1) as a functional target of marine natural product grincamycin B
Identification and characterization of isocitrate dehydrogenase 1 (IDH1) as a functional target of marine natural product grincamycin B
复制标题
异柠檬酸脱氢酶 1 (IDH1) 作为海洋天然产物格林霉素 B 功能靶点的鉴定和表征
DOI：
10.1038/s41401-020-0491-6
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发表时间：
2020-08-14
影响因子：
8.2
通讯作者：
Dang, Yong-jun
中科院分区：
文献类型：
--
作者：
Wang, Zheng;Li, Zeng-xia;Dang, Yong-jun
格林霉素（grincamycin, GCNs）是从放线菌和链霉菌中分离出来的一类抗肿瘤活性强的环霉素苷类化合物，但其抗肿瘤机制尚不清楚。在这项研究中，我们试图使用联合策略确定格林霉素B （GCN B）的细胞靶点，格林霉素B是最主要和最活跃的次级代谢产物之一。我们发现GCN b选择性诱导人急性早幼粒细胞白血病（APL）细胞系NB4凋亡是通过增加内质网应激和细胞内活性氧（ROS）积累。利用结合表型、转录组学和蛋白质微阵列方法的策略，我们发现异柠檬酸脱氢酶1（IDH1）是GCN B的假定靶点，并证实GCN是体外靶向野生型和突变型IDH1的选择性抑制剂的一个子集。众所周知，IDH1将异柠檬酸盐转化为2-氧葡萄糖酸盐（2-OG），维持细胞内2-OG的稳态。在NB4细胞和斑马鱼模型中验证了IDH1及其突变体作为GCN B的靶点。在NB4细胞中敲低IDH1引起的表型与GCN B处理相似，补充n -乙酰半胱氨酸部分挽救了IDH1干扰引起的NB4细胞凋亡。在斑马鱼模型中，GCN B能有效恢复IDH1（R132C）突变体过表达引起的髓系异常。综上所述，我们证明IDH1是GCNs的抗肿瘤靶点之一，这表明野生型IDH1可能是未来血液系统恶性肿瘤干预的潜在靶点。
Grincamycins (GCNs) are a class of angucycline glycosides isolated from actinomyceteStreptomycesstrains that have potent antitumor activities, but their antitumor mechanisms remain unknown. In this study, we tried to identify the cellular target of grincamycin B (GCN B), one of most dominant and active secondary metabolites, using a combined strategy. We showed that GCN B-selective-induced apoptosis of human acute promyelocytic leukemia (APL) cell line NB4 through increase of ER stress and intracellular reactive oxygen species (ROS) accumulation. Using a strategy of combining phenotype, transcriptomics and protein microarray approaches, we identified that isocitrate dehydrogenase 1(IDH1) was the putative target of GCN B, and confirmed that GCNs were a subset of selective inhibitors targeting both wild-type and mutant IDH1 in vitro. It is well-known that IDH1 converts isocitrate to 2-oxoglutarate (2-OG), maintaining intracellular 2-OG homeostasis. IDH1 and its mutant as the target of GCN B were validated in NB4 cells and zebrafish model. Knockdown of IDH1 in NB4 cells caused the similar phenotype as GCN B treatment, and supplementation ofN-acetylcysteine partially rescued the apoptosis caused by IDH1 interference in NB4 cells. In zebrafish model, GCN B effectively restored myeloid abnormality caused by overexpression of mutant IDH1(R132C). Taken together, we demonstrate that IDH1 is one of the antitumor targets of GCNs, suggesting wild-type IDH1 may be a potential target for hematological malignancies intervention in the future.