1-deoxysphingolipids bind to COUP-TF to modulate lymphatic and cardiac cell development

We identified 1-deoxysphingosine as a ligand for NR2F1 and NR2F2. To prove physiological relevance after in vitro binding assay we used different strategies to manipulate levels of 1-deoxysphingosines and canonical sphingosines in cardiomyocytes derived from human embrynonic stem cells. These manipulations included treating cells with sphingolipid biosynthesis inhibitors (myriocin), adding 1-deoxysphingosines to culture media or induced expression of the Sptlc1 HSAN mutant C133W to enhance 1-deoxysphingosine synthesis by cardiomyocytes. Examination of NR2F1 and NR2F2 targets after treatment with myriocin (sphingolipid biosynthetic inhibitor) or treatment with vehicle, canonical D-erythro-sphingosine or 1-deoxysphingosine. NR2F1 and NR2F2 targets were also analyzed after induction of 1-deoxysphingosine biosynthesis by tetracycline induced expression of SPTLC1 HSAN C133W mutant.

本研究鉴定出1-脱氧鞘氨醇（1-deoxysphingosine）可作为核受体亚家族2F成员1（NR2F1）与核受体亚家族2F成员2（NR2F2）的配体。为验证体外结合实验后的生理相关性，本研究采用多种策略调控人胚胎干细胞来源心肌细胞中1-脱氧鞘氨醇与经典鞘氨醇的水平。这些调控手段包括使用鞘脂生物合成抑制剂多球壳菌素（myriocin）、向培养基中添加1-脱氧鞘氨醇，或是诱导Sptlc1 HSAN突变体C133W的表达以增强心肌细胞的1-脱氧鞘氨醇合成能力。本研究分别检测了经多球壳菌素（鞘脂生物合成抑制剂）、溶剂对照、经典D-赤型鞘氨醇或1-脱氧鞘氨醇处理后的NR2F1与NR2F2靶基因表达情况。此外，本研究还分析了通过四环素诱导SPTLC1 HSAN C133W突变体表达以诱导1-脱氧鞘氨醇生物合成后的NR2F1与NR2F2靶基因表达情况。