PAR_1&2 driven placenta EVT invasion act via LRP5/6 as coreceptors

While the involvement of protease-activated receptors (PARs) in the physiological regulation of human placenta development, as in tumor biology, is recognized, the molecular pathway is unknown. We evaluated the impact of PAR₁ and PAR₂ function in cytotrophoblast (CTB) proliferation and invasion in a system of extravillous trophoblast (EVT) organ culture and in human cell-lines. Activation of PAR₁ - and PAR₂ -induced EVT invasion and proliferation, while the shRNA silencing of low-density lipoprotein receptor-related protein 5/6 (LRP5/6) inhibited these processes. PAR₁ and PAR₂ effectively induce β-catenin stabilization in a manner similar to that shown for the canonical β-catenin stabilization pathway yet independent of Wnts. Immunoprecipitation analyses and protein-protein docking demonstrated the co-association between either PAR₁ or PAR₂ with LRP5/6 forming an axis of PAR-LRP5/6-Axin. Noticeably, in PAR₁ -PAR₂ heterodimers a dominant role is assigned to PAR₂ over PAR₁ as shown by inhibition of PAR₁ -induced β-catenin levels, and Dvl nuclear localization. This inhibition takes place either by shRNA silenced hPar2 or in the presence of a TrPAR₂ devoid its cytoplasmic tail. Indeed, TrPAR₂ cannot form the PAR₁ -PAR₂ complex, obstructing thereby the flow of signals downstream. Elucidation of the mechanism of PAR-induced invasion contributes to therapeutic options highlighting key partners in the process.