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. Overall design: 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.