Poster Presentation Annual Meetings of the Endocrine Society of Australia and Society for Reproductive Biology and Australia and New Zealand Bone and Mineral Society 2016

Nuclear factor of activated T-cells is involved in regulation of inflammatory cytokines and soluble fms-like tyrosine kinase-1 production from human placenta (#457)

Louie Ye 1 , Fiona Brownfoot 1 , Amy Gratton 1 , Ping Cannon 1 , Natalie Hannan 1 , Stephen Tong 1 , Tu'uhevaha Kaitu'ulino
  1. Melbourne University, Heidelberg, VIC, Australia

INTRODUCTION: Preeclampsia (PE) is a serious complication of pregnancy characterized by poor placental establishment and placental hypoxia/ischemia, which leads to placental production of pro-inflammatory cytokines and anti-angiogenic factors such as soluble fms-like tyrosine kinase-1 (sFLT-1). However, the mechanisms regulating their production remain poorly understood.


AIM: The aim of this study was to characterize the expression of the nuclear factor of activated T-cells (NFAT) family of transcription factors (i.e. NFAT1-4) in preeclamptic placenta and determine their functional contribution to inflammatory cytokine and sFlt-1 production in primary placental cells.


METHODS: Archival preterm preeclamptic placental tissue (2 and expression of NFAT1-4 mRNA expression assessed. To determine the role of calcineurin/NFAT signaling in production of sFlt-1 and inflammatory cytokines, primary placental cells were isolated and treated with the inhibitor NFAT-calcineurin association-6 (INCA6). NFAT inhibition was assessed via inmmunofluorescent analysis of nuclear translocation, and effects on sFlt1 and inflammatory cytokines determined by ELISA and qPCR respectively.


RESULTS: mRNA expression of NFAT1-4 was not significantly altered in PE placenta relative to controls. Hypoxia significantly upregulated mRNA expression of NFAT1 (P<0.001) and NFAT3 (P<0.05) in primary human trophoblast. Inhibiting NFAT activity significantly reduced mRNA expression of FLT (P<0.001) and sflt-1 splice variant (e15a) (P<0.05), as well as mRNA expression of NFAT-dependent inflammatory cytokines such as IL-1b (P<0.01) and IL-10 (P<0.0001) in primary human trophoblast. Furthermore, inhibiting NFAT activity significantly reduced sFlt-1 protein production (P<0.05) from primary human trophoblast and explants in a dose-dependent manner.


CONCLUSIONS: In summary, we provide evidence that calcineurin-dependent NFAT proteins are hypoxia responsive and may be involved in the regulation of sFlt-1 and inflammatory cytokine production in preeclamptic placenta.