Lymphatic Endothelial Cells Derived Exosomes Promote Neolymphangiogenesis After Injury

Neolymphangiogenesis after myocardial infarction (MI) has been extensively documented; however the source of new lymphatic vessels (LVs) is still unknown. It has been described that new LVs may derive from resident lymphatic endothelial cells (LECs) or from trans-differentiation of endothelial cells (ECs) and pericytes from veins into LECs. Which signals activate both the spreading and the trans-differentiation needs to be investigated. LECs are known to release transcytotic vesicles and exosomes, thus, we hypnotized that the enriched protein and RNA cargo of LECs derived exosomes may promote the new LVs formation after injury. Exosomes derived from LECs were isolated from LECs cultured conditioned medium and HUVECs were treated with different concentration of exosomes cargo for 10 days. After treatment, q-PCR for the major endothelial and lymphatic endothelial markers was performed on treated HUVECs. In a dose dependent manner we observed a decreased expression of KDR and CD31, endothelial markers that are not expressed by LECs and an increased expression of Prox-1 and VEGFR3. Prox-1 is the major lymphatic endothelial transcription factor that is not activated in ECs and is responsible for the transcription of LYVE-1 and podoplanin. VEGFR3 is the only known vascular endothelial growth factor receptor expressed on LECs which is not displayed by ECs. Next, we evaluated the effect of LECs derived exosomes on in vitro lymphangiogenesis assay by measuring the ability of LECs to organize into capillary-like structures on matrigel. LECs treated with LECs derived exosomes are able to form tubes and branches as early as within 1 hour compared to untreated cells. Our preliminary results show cargo of exosomes from LECs activates the signal transduction and metabolism of LECs and HUVECs promoting the new LVs formation through spreading and trans-differentiation of the targeted cells.