在各种细胞中
The sphingosine kinase 2 inhibitors ABC294640 and K145 elevate (dihydro)sphingosine 1-phosphate levels in various cells
影响因子:4.10000
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
摘要
产生生物活性脂质二氢磷酸盐1-磷酸盐（DHS1P）和1-磷酸盐（S1P）的生物活性脂质二氢磷酸盐的酶（SPHKS）与包括癌症和感染在内的各种疾病有关。因此，已经开发了许多SPHK抑制剂。尽管已经描述了针对选定剂的靶向效应，但SPHK抑制剂主要用于研究中，而无需监测对鞘脂组的影响。现在，我们已经研究了七种常用的SPHK抑制剂（5C，ABC294640（Opaganib），N，N，N-二甲基肾上腺素，K145，PF-543，SLM6031434和SKI-II）对Changolipids在Changolipids changolipids changolipids changolipids changolipial的概况的影响。虽然我们观察到N，N-二甲基肾上腺素，PF-543，SKI-II和SLM6031434的预期（DH）S1P降低（DH）降低几乎没有任何影响。值得注意的是，对于K145和ABC294640（据报道都是SPHK2），我们观察到跨细胞系中DHS1P和S1P的剂量依赖性强大增加。 SPHK1的补偿作用可以排除，因为该观察结果也是SPHK1缺乏的HK-2细胞。此外，我们观察到对所有测试的所有抑制剂的二氢氧酰胺去饱和酶活性的影响，如前所述，对于ABC294640和SKI-II。在其他机械研究中，我们更详细地研究了用ABC294640和K145的短期细胞处理后DHS1P和S1P的大量增加。我们发现，两种化合物都会影响从头合成的鞘脂，并以3-酮二氢还原酶和二氢氧疗法的去饱和酶作为其靶标。 我们的研究表明，所测试的七个SPHK抑制剂中没有一个没有意外的目标和/或靶向效果。因此，在机械研究中使用SPHK抑制剂时，监测细胞鞘脂谱很重要。
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.