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A novel technique to characterize procoagulant platelet formation and evaluate platelet procoagulant tendency in cats
Meg Shaverdian1, Nghi Nguyen1, Ronald H L Li2
1Department of Surgical and Radiological Sciences, School of Veterinary Medicine, University of California, Davis, Davis, CA, United States.
Insights
This study developed a flow cytometry method to assess procoagulant platelets in cats, identifying thrombin and convulxin as key agonists. This research offers insights into feline hypertrophic cardiomyopathy complications.
Area of Science:
- Veterinary Medicine
- Cardiovascular Research
- Hematology
Background:
- Cardiogenic arterial thromboembolism (CATE) is a severe complication of hypertrophic cardiomyopathy (HCM) with high mortality.
- Platelets are critical in hemostasis and CATE progression, with procoagulant platelets playing a significant role.
- Studying feline procoagulant platelets is challenging due to species-specific physiology.
Purpose of the Study:
- To develop a flow cytometry method for assessing procoagulant platelet markers in cats.
- To identify agonists that induce procoagulant platelet phenotypes in felines.
- To establish a scoring system for evaluating platelet procoagulant tendency in cats.
Main Methods:
- Platelets from healthy cats were isolated and activated with thrombin, collagen (COL), and convulxin (CVX).
- Flow cytometry was used to sequentially measure mitochondrial membrane potential (ΔΨm) loss, phosphatidylserine (PS) externalization, and P-selectin upregulation.
- A novel scoring system was implemented to quantify the procoagulant tendency.
Main Results:
- Thrombin with CVX significantly reduced ΔΨm and increased P-selectin expression.
- Thrombin with COL and CVX significantly enhanced PS externalization.
- Thrombin and CVX yielded the highest procoagulant tendency scores, with 75% of cats scoring ≥2/3.
Conclusions:
- Sequential flow cytometry is an effective technique for studying procoagulant platelets in cats.
- This method can help understand CATE in feline HCM.
- Identifying key agonists and markers aids in potential diagnostic and therapeutic strategies.
Introduction:
Cardiogenic arterial thromboembolism (CATE) is a life-threatening complication of hypertrophic cardiomyopathy (HCM) with a high mortality rate. As the primary responders in hemostasis, platelets play a crucial role in the progression of CATE. Procoagulant platelets are a subpopulation of activated platelets that facilitate thrombin generation to strengthen thrombus structure. Since their discovery, procoagulant platelets have gained a considerable amount of attention due to their potential role in thromboembolic complications. Uncovering the key phenotypic markers and agonists underlying the procoagulant response in feline platelets may provide diagnostic and therapeutic insights in the treatment of CATE. However, species differences in platelet physiology and the sensitive nature of feline platelets pose some significant challenges in studying procoagulant platelets in cats.
Objectives:
To first devise a flow cytometric method to sequentially assess procoagulant platelet markers and to identify agonists that could induce procoagulant platelet phenotypes. Furthermore, a novel scoring system was established to evaluate the procoagulant tendency of platelets in cats.
Methods:
Platelets were isolated from eight healthy cats and activated by thrombin in the presence or absence of collagen (COL) and convulxin (CVX). The following markers were measured sequentially by flow cytometry: (1) loss of mitochondrial membrane potential (ΔΨm), (2) phosphatidylserine (PS) externalization, and (3) P-selectin upregulation.
Results:
Thrombin in the presence of CVX significantly reduced ΔΨm and induced P-selectin upregulation (p = 0.0078, p = 0.0128, respectively). In addition, thrombin-treated platelets in the presence of COL and CVX augmented PS externalization significantly (p = 0.043, p = 0.0172, respectively). Of the agonists tested, thrombin and CVX resulted in the highest procoagulant tendency score with 75% cats reaching a score ≥2/3. The number of cats with a procoagulant tendency score of 2 or 3 for thrombin and thrombin + COL was 4/8 (50%) and 5/8 (62.5%), respectively.
Discussion:
Sequential analysis of procoagulant markers via flow cytometry may be an effective technique for studying procoagulant platelets in cats.
