Head-to-Head Comparison of Consensus-Recommended Platelet Function Tests to Assess P2Y12 Inhibition—Insights for Multi-Center Trials

Abstract

The vasodilator-associated stimulated phosphoprotein (VASP) phosphorylation level is a highly specific method to assess P2Y12 receptor inhibition. Traditionally, VASP phosphorylation is analyzed by flow cytometry, which is laborious and restricted to specialized laboratories. Recently, a simple ELISA kit has been commercialized. The primary objective of this study was to compare the performance of VASP assessment by ELISA and flow cytometry in relation to functional platelet aggregation testing by Multiplate® whole-blood aggregometry. Blood from 24 healthy volunteers was incubated with increasing concentration of a P2Y12 receptor inhibitor (AR-C 66096). Platelet function testing was carried out simultaneously by Multiplate® aggregometry and by VASP assessment through ELISA and flow cytometry. As expected, increasing concentrations of the P2Y12 receptor inhibitor induced a proportional inhibition of platelet aggregation and P2Y12 receptor activation across the modalities. Platelet reactivity index values of both ELISA- and flow cytometry-based VASP assessment methods correlated strongly (r = 0.87, p < 0.0001) and showed minimal bias (1.05%). Correlation with Multiplate® was slightly higher for the flow cytometry-based VASP assay (r = 0.79, p < 0.0001) than for the ELISA-based assay (r = 0.69, p < 0.0001). Intraclass correlation (ICC) was moderate for all the assays tested (ICC between 0.62 and 0.84). However, categorization into low, optimal, or high platelet reactivity based on these assays was strongly concordant (κ between 0.86 and 0.92). In conclusion, the consensus-recommended assays with their standardized cut-offs should not be used interchangeably in multi-center clinical studies but, rather, they should be standardized throughout sites.

Extracellular adenosine 5ʹ-diphosphate promotes MCP-1/CCL2 expression via the P2Y13 purinergic receptor/ERK signaling axis in temporomandibular joint-derived mouse fibroblast-like synoviocytes

“…2 B). Bélanger et al reported that ARC-66096 (10 µM) significantly inhibits platelet aggregation [ 19 ], and Quintas et al demonstrated that ARC-66096 (10 µM) can significantly abrogate adenosine 5ʹ- O -(2-thio)-diphosphate-induced astroglial proliferation [ 20 ], thus indicating 10 µM to be the optimal concentration of ARC-66096 for antagonizing ADP-activated P2Y 12 signaling. Notably, we established that the P2Y 13 antagonist MRS2211 (50–100 µM) significantly abrogated the ADP-promoted expression of MCP-1/CCL2 in a concentration-dependent manner (Fig.…”

Extracellular adenosine 5ʹ-diphosphate promotes MCP-1/CCL2 expression via the P2Y13 purinergic receptor/ ERK signaling axis in temporomandibular joint-derived mouse fibroblast-like synoviocytes

“…2B). Bélanger et al reported that ARC-66096 (10 µM) signi cantly inhibits platelet aggregation [19], and Quintas et al demonstrated that ARC-66096 (10 µM) can signi cantly abrogate adenosine 5 -O-(2-thio)-diphosphate-induced astroglial proliferation [20], thus indicating 10 µM to be the optimal concentration of ARC-66096 for antagonizing ADP-activated P2Y 12 signaling. Notably, we established that the P2Y 13 antagonist MRS2211 (50-100 µM) signi cantly abrogated the ADP-promoted expression of MCP-1/CCL2 in a concentration-dependent manner (Fig.…”

Perioperative platelet reactivity over time in patients undergoing vascular surgery: An observational pilot study

“…The VASP assay was performed according to the manufacturer’s instructions (Diagnostica Stago S.A.S.) [ 8 ]. Results were obtained by flow cytometry and expressed as platelet reactivity index (PRI).…”