Ceramide 1 Phosphate Is a Potent Chemoattractant Factor of Endothelial Colony Forming Cells and Improve Post Ischemia Tissue Regeneration

ZUBIRY PR; BOISSON-VIDAL, C; MENA HA; SCHATTNER M; DIZIER B; NEGROTTO S

Melbourne

Congreso; XXVII International Society on Thrombosis and Haemostasis (ISTH) Congress and 65th Annual Scientific and Standardization Committee (SSC) Meeting; 2019

International Society on Thrombosis and Haemostasis

Background : Ceramide 1 phosphate (C1P) is a well- known chemo-taxis inductor in macrophages and murine progenitor cells. In en-dothelial colony forming cells (ECFC), we have shown that C1P improved proliferation and tubule formation, although C1P chemot-actic effects remain unknown. Aims : Considering that, C1P levels are elevated in ischemic tissues and that ECFC migration is a key step post- ischemia tissue revascu-larization, we here aimed to study whether C1P is an ECFC chemoat-tractant factor and improve their revascularization abilities in vivo.Methods : Human cord blood- derived CD34 + cells were cultured in EGM2 and, after 14- 18 days, ECFC colonies were obtained. ECFC were treated with C1P short chain analog C8- C1P. N=3- 6, *p< 0.05, one- way ANOVA. Results : We found that C8- C1P is a potent chemoattractant factor for ECFC not only per se , but also combined with SDF- 1 (transwells, Figure 1A). C8- C1P- induced chemotaxis was completely sup-pressed by pharmacological inhibitors of ERK1/2 and AKT pathways (Figure 1B). In vivo , we observed that C8- C1P not only has a potent vasculogenic effect by itself, but also potentiated plug vasculariza-tion mediated by ECFC (Geltrex plug implants, Figure 2A). Moreover, in a murine model of hind limb ischemia, intramuscular injection of C8- C1P enhanced blood perfusion in the ischemic limb and slightly increased the revascularization mediated by untreated ECFC trans-plantation. Furthermore, administration of C1P- pretreated ECFC to-gether with intramuscular C1P resulted in a significant improvement of leg reperfusion compared to each condition alone (Figure 2B). Conclusions : In conclusion, C8- C1P induce ECFC chemotaxis in vitro , through AKT and ERK1/2 activation, and in vivo in a hind limb ischemia model, where C8- C1P not only attract ECFC to the ischemic muscle, but also augmented ECFC revascularization abilities. Our results highlight the therapeutic potential of C1P to improve post ischemia tissue regeneration.