鞘氨醇激酶2抑制剂ABC294640和K145在多种细胞中升高（二氢）鞘氨醇-1-磷酸水平
The sphingosine kinase 2 inhibitors ABC294640 and K145 elevate (dihydro)sphingosine 1-phosphate levels in various cells
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影响因子:4.1
分区:医学2区 / 生化与分子生物学2区
发表日期:2024 Oct
作者:
Agata Prell, Dominik Wigger, Andrea Huwiler, Fabian Schumacher, Burkhard Kleuser
DOI:
10.1016/j.jlr.2024.100631
摘要
鞘氨醇激酶（SphKs）是一类产生具有生物活性的脂质二氢鞘氨醇-1-磷酸（dhS1P）和鞘氨醇-1-磷酸（S1P）的酶，涉及多种疾病，包括癌症和感染。因此，已经开发出多种SphK抑制剂。虽然某些药物存在非靶向作用，但大多在研究中使用，未监测其在鞘脂组分中的作用。我们目前研究了七种常用的SphK抑制剂（5c、ABC294640（奥帕尼比）、N,N-二甲基鞘氨醇、K145、PF-543、SLM6031434 和 SKI-II）在Chang、HepG2和人脐静脉内皮细胞中的鞘脂谱变化。虽然观察到N,N-二甲基鞘氨醇、PF-543、SKI-II和SLM6031434导致的预期的（dh）S1P减少，但5c几乎没有影响。令人惊讶的是，在被认为特异性作用于SphK2的K145和ABC294640中，我们观察到在细胞系中剂量依赖性显著增加的dhS1P和S1P水平。这一现象排除了SphK1的补偿作用，因为在SphK1缺乏的HK-2细胞中亦观察到类似变化。此外，所有抑制剂均影响二氢酰胺脱氢酶活性，之前已报道ABC294640和SKI-II具有此作用。在机制研究中，我们详细分析了ABC294640和K145短期处理后引起的dhS1P和S1P大量升高的原因。发现这两种化合物影响鞘脂的de novo合成，其靶点为3-酮二氢鞘氨醇还原酶和二氢酰胺脱氢酶。我们的研究表明，所测试的七种SphK抑制剂都存在意外的靶向和/或非靶向作用，因此在机制研究中，应监测细胞鞘脂的变化。
Abstract
Sphingosine kinases (SphKs), enzymes that produce the bioactive lipids dihydrosphingosine 1-phosphate (dhS1P) and sphingosine 1-phosphate (S1P), are associated with various diseases, including cancer and infections. For this reason, a number of SphK inhibitors have been developed. Although off-target effects have been described for selected agents, SphK inhibitors are mostly used in research without monitoring the effects on the sphingolipidome. We have now investigated the effects of seven commonly used SphK inhibitors (5c, ABC294640 (opaganib), N,N-dimethylsphingosine, K145, PF-543, SLM6031434, and SKI-II) on profiles of selected sphingolipids in Chang, HepG2, and human umbilical vein endothelial cells. While we observed the expected (dh)S1P reduction for N,N-dimethylsphingosine, PF-543, SKI-II, and SLM6031434, 5c showed hardly any effect. Remarkably, for K145 and ABC294640, both reported to be specific for SphK2, we observed dose-dependent strong increases in dhS1P and S1P across cell lines. Compensatory effects of SphK1 could be excluded, as this observation was also made in SphK1-deficient HK-2 cells. Furthermore, we observed effects on dihydroceramide desaturase activity for all inhibitors tested, as has been previously noted for ABC294640 and SKI-II. In additional mechanistic studies, we investigated the massive increase of dhS1P and S1P after short-term cell treatment with ABC294640 and K145 in more detail. We found that both compounds affect sphingolipid de novo synthesis, with 3-ketodihydrosphingosine reductase and dihydroceramide desaturase as their targets. Our study indicates that none of the seven SphK inhibitors tested was free of unexpected on-target and/or off-target effects. Therefore, it is important to monitor cellular sphingolipid profiles when SphK inhibitors are used in mechanistic studies.