Anti-inflammatory and pro-angiogenic effects of placental mesenchymal stromal cells conditioned media on preeclamptic placental tissue.

Abstract

s / Placenta 35 (2014) A1eA112 A87 increasing doses (0-100mM) on the following outputs 1) heme-oxygenase1 expression (HO-1, key antioxidant enzyme) 2) production of sFlt1 and sEng and 3) assays of endothelial dysfunction 4) endothelial tube formation. Endothelial dysfunction was modeled by adding TNFa or preeclamptic (PE) patient serum (to induce dysfunction) and subsequently adding PPIs. Results: All PPIs induced a significant dose dependent increase in HO-1 mRNA and protein (average 4-5 fold increase). Furthermore, PPIs induced nuclear translocation of Nrf2 (transcription factor). PPIs caused a significant dose dependent reduction in mRNA expression of sFlt1 variants (e15a and i13), and reduced secretion of both sFlt1 and sEng (dose dependently) in all three cell types. PPIs rescued both TNFa and PE serum induced endothelial dysfunction (evidenced by increased VCAM1 and endothelin-1 expression, but this was significantly attenuated by adding PPIs (decreased VCAM and ET-1 mRNA and protein levels). Finally, PPIs reversed TNFainduced disruption of in vitro endothelial cell tube formation. Summary and conclusions: In primary human tissues, PPIs potently 1) induced HO-1, 2) reduced sFlt1 and sEng secretion and 3) quenched endothelial dysfunction 4) rescued endothelial tube formation. PPIs may represent a highly novel candidate therapeutic for severe preeclampsia. Given PPIs are safe in pregnancy, they could be fast-tracked to clinical trials. P2.81-N. ANTI-INFLAMMATORY AND PRO-ANGIOGENIC EFFECTS OF PLACENTAL MESENCHYMAL STROMAL CELLS CONDITIONED MEDIA ON PREE CLAMPTIC PLACENTAL TISSUE. Anna Maria Nuzzo, Domenica Giuffrida, Ettore Piccoli, Cristian Zenerino, Rossella Barrile, Tullia Todros, Alessandro Rolfo Dept. of Surgical Sciences, University of Turin, Turin, Italy Objective: Placenta-derived Mesenchymal Stromal Cells (PDMSCs) are renowned for their unique anti-inflammatory and pro-angiogenic trophic activities. Indeed, we hypothesized that PDMSCs-derived molecules could counteract the production of pro-inflammatory cytokines and antiangiogenic factors typical of preeclamptic (PE) placentae. To verify our hypothesis, herein we investigated the expression of Macrophage-migration Inhibitory Factor (MIF), Tumor Necrosis Factor a (TNF-a) and Interleukin 6 (IL-6) cytokines, and anti-angiogenic soluble Fms-like tyrosine kinase-1 (sFlt1) in PE villous explants treated by PDMSCs conditioned media (CM). Methods: PDMSCs were isolated from control (n1⁄430) and PE (n1⁄430) placentae. At passage 5, control and PE-PDMSC were plated (1x105 cells/ ml) in DMEM without FBS. Conditioned media (CM) were collected after 48h. PE (n1⁄448) villous explants were treated for 72h using control PDMSCs CM. MIF, TNF-a, IL-6 and sFlt1 mRNA expression were evaluated by Real Time PCR. Results:Normal PDMSCs CM treatment significantly decreasedMIF, TNF-a, IL-6 and sFlt-1 mRNA levels (p<0.05, 3 Fold decrease) in PE villous explants relative to untreated controls. Conclusion: Our results suggest that PDMSCs CM could be used to neutralize the exacerbated inflammation and impaired angiogenesis typical of PE placentae. These data open to the development of innovative PDMSCs-based therapeutic tools for the treatment of preeclampsia. P2.82. A COMBINATION OF SINGLE NUCLEOTIDE POLYMORPHISMS IN THE 3’UNTRANSLATED REGION OF HLA-G IS ASSOCIATED WITH PREECLAMPSIA Kevin Quach , Stephanie Grover , Shlomit Kenigsberg , Clifford Librach a,b,c CReATe Fertility Centre, Toronto, Ontario, Canada; Women’s College Hospital, Toronto, Ontario, Canada; University of Toronto, Toronto,