FEAM: A NOVEL MODULATOR FOR NEUROINFLAMMATION
- Efrat Shavit Stein, Israel
Abstract
Background and Aims
Neural inflammation is regulated by coagulation proteins including activated protein C (aPC) and its endothelial protein c receptor (EPCR) which together activate protease activated receptor 1 (PAR1) inducing anti-inflammatory effects. We have synthesized a novel molecule based on the binding site of FVII/aPC to EPCR (FEAM) and studied its effectiveness in the treatment of neuroinflammation.
Methods
An in- vitro model for neuroinflammation was induced by LPS applied to N9 microglia cells. In-vivo neuroinflammation was induced by LPS systemic injection to ICR mice and behavior was assessed by the stair-case test. Thrombin and aPC activity from cells and brains were measured by enzymatic fluorescence assays. Proliferation was measured by XTT activity assay. Coagulation factors and inflammation markers levels were evaluated by western blot and real-time PCR.
Results
FEAM prevented the LPS induced increased proliferation rate (1 vs 1.5 arbitrary units (aU), p<0.001) and PAR1 expression in N9 (1.7 vs. 1.2, p<0.001). FEAM also prevented the decreased aPC activity induced by LPS (0.46 vs 0.62 aU, p<0.003) and prevented the elevation of coagulation factors (FX and thrombin) and inflammatory markers (TNFα). In the whole animal model FEAM prevented the LPS induced elevated brain thrombin activity and other coagulation and inflammation factors. FEAM treatment induced improvement in general health indices such as weight, learning and memory and mobility.
Conclusions
In conclusion, FEAM modulation of the FVII-aPC-EPCR pathway may shift the thrombin/PAR1 pathway toward aPC-EPCR mediated protective downstream effects.