Protein disulphide isomerase and NADPH oxidase 1 cooperate to control platelet function and are associated with cardiometabolic disease risk factors

Simoes Gaspar, R., Sage, T., Little, G., Kriek, N., Pula, G. and Gibbins, J. M. ORCID: https://orcid.org/0000-0002-0372-5352 (2021) Protein disulphide isomerase and NADPH oxidase 1 cooperate to control platelet function and are associated with cardiometabolic disease risk factors. Antioxidants, 10 (3). 497. ISSN 2076-3921 doi: 10.3390/antiox10030497

Abstract/Summary

Background: Protein disulphide isomerase (PDI) and NADPH oxidase 1 (Nox- 1) regulate platelet function and reactive oxygen species (ROS) generation, suggesting potentially interdependent roles. Increased platelet reactivity and ROS production have been correlated with cardiometabolic disease risk factors. Objectives: To establish whether PDI and Nox-1 cooperate to control platelet function. Methods: Immunofluo- rescence microscopy was utilised to determine expression and localisation of PDI and Nox-1. Platelet aggregation, fibrinogen binding, P-selectin exposure, spreading and cal- cium mobilization were measured as markers of platelet function. A cross-sectional population study (n=136) was conducted to assess the relationship between platelet PDI and Nox-1 levels and cardiometabolic risk factors. Results: PDI and Nox-1 co-localized upon activation induced by the collagen receptor GPVI. Co-inhibition of PDI and Nox-1 led to additive inhibition of GPVI-mediated platelet aggregation, activation and calcium flux. This was confirmed in murine Nox-1-/- platelets treated with PDI inhibitor be- pristat, without affecting bleeding. PDI and Nox-1 together contributed to GPVI signal- ling that involved the phosphorylation of p38 MAPK, p47phox, PKC and Akt. Platelet PDI and Nox-1 levels were upregulated in obesity, with platelet Nox-1 also elevated in hypertensive individuals. Conclusions: We show that PDI and Nox-1 cooperate to con- trol platelet function and are associated with cardiometabolic risk factors.