Discovering Anti-platelet Drug Combinations with an Integrated Model of Activator-Inhibitor Relationships, Activator-Activator Synergies and Inhibitor-Inhibitor Synergies
Fig 2
Heatmap of platelet activation (log ADP release) in each donor for each reagent combination.
Columns: 10 donors. Rows: different experimental conditions. Green: activated platelets with high ADP release, measured in log10 Arbitrary Absorbance Units (AAU); red: non-activated platelets. White vertical line: actual value of log10 (AAU). The white vertical dashed lines across each column represent the middle value between the maximum and minimum values observed for the entire dataset. Data were grouped by hierarchical clustering. Any technically replicated results were represented by their means. The five activators used were used at doses typically corresponding to their EC50 (see text): 0.025 μM Epinephrine (Ea), 0.5 μM U46619 (Xa), 1 μg/ml CRP (Ca), 4 μM TRAP (Ta), and 10 μM ADP (Aa), respectively intended to activate the epinephrine, thromboxane, collagen, thrombin and ADP receptors; K represents a cocktail comprising all five activators combined at a dilution corresponding to their combined EC50 (the individual concentrations shown, multiplied by 0.1636). The five inhibitors used at their IC50 values were 1uM Yohimbine (Ei), 68.39 nM SQ29548 (Xi), 16.5 nM Wortmannin (Pi), 2.85 uM BMS200261 (Ti), and 36.77 uM MRS2395 (Ai), respectively intended to inhibit the epinephrine receptor, thromboxane receptor, PI3K, thrombin receptor and ADP receptor. For comparison purposes, the double doses of individual activators and inhibitors were included, which are shown preceded by the number “2”; EC90 and IC90 doses (see text) were also included for comparison, with the prefix “90”.