Interspecies comparison of simultaneous thrombin and plasmin generation

Abstract

Animal models of hemostasis are often extrapolated to humans; however, only a few studies have compared coagulation and fibrinolysis across species. Simultaneous thrombin (TG) and plasmin (PG) generation is useful to assessing coagulation and fibrinolysis within the same sample. In this study, we performed simultaneous TG and PG analysis in blood plasma samples from humans and 6 species commonly evaluated in pre-clinical research. TG and PG were investigated in male and female donor platelet-poor plasmas (PPP) obtained from 28 healthy humans, 10 baboons, 12 rhesus monkeys, 20 Yorkshire pigs, 20 Sprague-Dawley rats, 10 New Zealand White rabbits and 14 Hartley guinea pigs. The continuous generation of the 7-amino-4-methylcoumarin (AMC) from substrates specific to thrombin or plasmin was monitored. The thrombin and plasmin concentration peak heights (PH) and production rates (PR) were calculated. TG and PG parameters from baboon and rhesus macaque plasma approximated that of humans. The other species differed significantly from both human and nonhuman primates. For example, swine and rat plasmas demonstrated similar TG, but swine plasmas did not generate plasmin. TG and PG parameters from Guinea pig samples were extremely low, while rabbit plasmas showed variable PG curves demonstrating one or two peaks with low and high PR values, respectively. Correlations between PH and PR values were significant with the exceptions of human PG, baboon TG, rat TG and Guinea pig PG. These findings are informative to pre-clinical animal species selection and optimization of coagulation and fibrinolysis translational research.Animal models of hemostasis and thrombosis are widely used in basic and pharmaceutical research 1 . Results are frequently extrapolated to humans; however, data obtained from animal models often falls short of accurate predictions in human response 2 . Further, assays have limited relevant comparisons of hemostasis in human blood with that of differing animal species. The novel global assays of hemostasis such as thrombin generation (TG) and thromboelastography (TEG) or thromboelastometry (ROTEM) provide an opportunity to assess similarities in coagulation and fibrinolysis ex vivo on blood obtained from different species. Although TG and thromboelastographic assays are often used for analysis of hemostasis in different animals 1,3-9 , comparison among different species is rarely made. There are limited studies that compare blood coagulation and fibrinolysis in various animals under the same conditions 10-17 . TG assays are sensitive for use in clinical and basic research studies; however, fibrinolysis testing primarily focused on clot lysis time, is often insufficiently sensitive and demonstrates high data variability 8,9,11 . Since 2011, several hemostasis assays have focused on simultaneous registration of thrombin and plasmin generation (PG) in plasma using tissue factor (TF) and tissue plasminogen activator (tPA), respectively 18-21 . These approaches suggest high sensitivity to assessment of coagulation an...

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