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Article ; Online: Whole Blood Procoagulant Platelet Flow Cytometry Protocol for Heparin-Induced Thrombocytopenia (HIT) and Vaccine-Induced Immune Thrombotic Thrombocytopenia (VITT) Testing.

Lee, Christine S M / Powell, Matthew C / Chen, Vivien M

Methods in molecular biology (Clifton, N.J.)

2023 Volume 2663, Page(s) 441–461

Abstract: Heparin-induced thrombocytopenia (HIT) is a well-characterized, iatrogenic complication of heparin anticoagulation with significant morbidity. In contrast, vaccine-induced immune thrombotic thrombocytopenia (VITT) is a recently recognized severe ... ...

Abstract	Heparin-induced thrombocytopenia (HIT) is a well-characterized, iatrogenic complication of heparin anticoagulation with significant morbidity. In contrast, vaccine-induced immune thrombotic thrombocytopenia (VITT) is a recently recognized severe prothrombotic complication of adenoviral vaccines, including the ChAdOx1 nCoV-19 (Vaxzevria, AstraZeneca) and Ad26.COV2.S (Janssen, Johnson & Johnson) vaccines against COVID-19. The diagnosis of HIT and VITT involve laboratory testing for antiplatelet antibodies by immunoassays followed by confirmation by functional assays to detect platelet-activating antibodies. Functional assays are critical to detect pathological antibodies due to the varying sensitivity and specificity of immunoassays. This chapter presents a protocol for a novel whole blood flow cytometry-based assay to detect procoagulant platelets in healthy donor blood in response to plasma from patients suspected of HIT or VITT. A method to identify suitable healthy donors for HIT and VITT testing is also described.
MeSH term(s)	Humans ; Blood Platelets ; Ad26COVS1 ; COVID-19 Vaccines/adverse effects ; ChAdOx1 nCoV-19 ; Flow Cytometry ; COVID-19 ; Thrombocytopenia/chemically induced ; Thrombocytopenia/diagnosis ; Thrombosis ; Antibodies ; Vaccines ; Platelet Factor 4
Chemical Substances	Ad26COVS1 (JT2NS6183B) ; COVID-19 Vaccines ; ChAdOx1 nCoV-19 (B5S3K2V0G8) ; Antibodies ; Vaccines ; Platelet Factor 4 (37270-94-3)
Language	English
Publishing date	2023-05-18
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ISSN	1940-6029
ISSN (online)	1940-6029
DOI	10.1007/978-1-0716-3175-1_29
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Article ; Online: Mechanisms of Thrombosis in Heparin-Induced Thrombocytopenia and Vaccine-Induced Immune Thrombotic Thrombocytopenia.

Selvadurai, Maria V / Favaloro, Emmanuel J / Chen, Vivien M

Seminars in thrombosis and hemostasis

2023 Volume 49, Issue 5, Page(s) 444–452

Abstract: Heparin-induced thrombocytopenia (HIT) and vaccine-induced immune thrombotic thrombocytopenia (VITT) are rare, iatrogenic immune-mediated conditions with high rates of thrombosis-related morbidity and mortality. HIT is a long-recognized reaction to the ... ...

Abstract	Heparin-induced thrombocytopenia (HIT) and vaccine-induced immune thrombotic thrombocytopenia (VITT) are rare, iatrogenic immune-mediated conditions with high rates of thrombosis-related morbidity and mortality. HIT is a long-recognized reaction to the administration of the common parenterally administered anticoagulant heparin (or its derivatives), while VITT is a new, distinct syndrome occurring in response to adenovirus-based vaccines against coronavirus disease 2019 and potentially other types of vaccines. A feature of both HIT and VITT is paradoxical thrombosis despite a characteristic low platelet count, mediated by the presence of platelet-activating antibodies to platelet factor 4. Several additional factors have also been suggested to contribute to clot formation in HIT and/or VITT, including monocytes, tissue factor, microparticles, endothelium, the formation of neutrophil extracellular traps, complement, procoagulant platelets, and vaccine components. In this review, we discuss the literature to date regarding mechanisms contributing to thrombosis in both HIT and VITT and explore the pathophysiological similarities and differences between the two conditions.
MeSH term(s)	Humans ; COVID-19 ; Thrombocytopenia ; Purpura, Thrombocytopenic, Idiopathic ; Vaccines ; Thrombosis ; Platelet Factor 4
Chemical Substances	Vaccines ; Platelet Factor 4 (37270-94-3)
Language	English
Publishing date	2023-01-27
Publishing country	United States
Document type	Review ; Journal Article
ZDB-ID	196901-8
ISSN	1098-9064 ; 0094-6176
ISSN (online)	1098-9064
ISSN	0094-6176
DOI	10.1055/s-0043-1761269
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Article: Mechanisms of Thrombosis in Heparin-Induced Thrombocytopenia and Vaccine-Induced Immune Thrombotic Thrombocytopenia

Selvadurai, Maria V. / Favaloro, Emmanuel J. / Chen, Vivien M.

Seminars in Thrombosis and Hemostasis

(Editorial Compilation—XIII)

2023 Volume 49, Issue 05, Page(s) 444–452

Series title	Editorial Compilation—XIII
Abstract	Heparin-induced thrombocytopenia (HIT) and vaccine-induced immune thrombotic thrombocytopenia (VITT) are rare, iatrogenic immune-mediated conditions with high rates of thrombosis-related morbidity and mortality. HIT is a long-recognized reaction to the administration of the common parenterally administered anticoagulant heparin (or its derivatives), while VITT is a new, distinct syndrome occurring in response to adenovirus-based vaccines against coronavirus disease 2019 and potentially other types of vaccines. A feature of both HIT and VITT is paradoxical thrombosis despite a characteristic low platelet count, mediated by the presence of platelet-activating antibodies to platelet factor 4. Several additional factors have also been suggested to contribute to clot formation in HIT and/or VITT, including monocytes, tissue factor, microparticles, endothelium, the formation of neutrophil extracellular traps, complement, procoagulant platelets, and vaccine components. In this review, we discuss the literature to date regarding mechanisms contributing to thrombosis in both HIT and VITT and explore the pathophysiological similarities and differences between the two conditions.
Keywords	heparin-induced thrombocytopenia ; vaccine-induced thrombotic thrombocytopenia ; thrombosis ; platelet factor 4
Language	English
Publishing date	2023-01-27
Publisher	Thieme Medical Publishers, Inc.
Publishing place	Stuttgart ; New York
Document type	Article
ZDB-ID	196901-8
ISSN	1098-9064 ; 0094-6176
ISSN (online)	1098-9064
ISSN	0094-6176
DOI	10.1055/s-0043-1761269
Database	Thieme publisher's database

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Article: Perspective: Collagen induced platelet activation

Pennings, Gabrielle J / Reddel, Caroline J / Chen, Vivien M / Gnanenthiran, Sonali R / Kritharides, Leonard

Frontiers in cardiovascular medicine

2023 Volume 9, Page(s) 1104744

Abstract: Colchicine has been demonstrated to reduce cardiovascular death, myocardial infarction (MI), ischemic stroke, and ischemia-driven coronary revascularization in people with coronary artery disease (CAD). These reductions were observed even in patients ... ...

Abstract	Colchicine has been demonstrated to reduce cardiovascular death, myocardial infarction (MI), ischemic stroke, and ischemia-driven coronary revascularization in people with coronary artery disease (CAD). These reductions were observed even in patients already taking antiplatelet therapy. As well as having anti-inflammatory effects, colchicine demonstrates antiplatelet effects. We propose that colchicine's antiplatelet effects primarily target collagen-induced platelet activation
Language	English
Publishing date	2023-01-19
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2781496-8
ISSN	2297-055X
ISSN	2297-055X
DOI	10.3389/fcvm.2022.1104744
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Article ; Online: Preventability of venous thromboembolism in hospitalised patients.

Narayan, Sujita W / Gad, Fady / Chong, Julianne / Chen, Vivien M / Patanwala, Asad E

Internal medicine journal

2022 Volume 53, Issue 4, Page(s) 577–583

Abstract: Background: Hospital-acquired venous thromboembolism (VTE) is a major cause of morbidity and mortality.: Aims: To determine the proportion of patients with hospital-acquired VTE that are preventable.: Methods: This was a retrospective study of ... ...

Abstract	Background: Hospital-acquired venous thromboembolism (VTE) is a major cause of morbidity and mortality. Aims: To determine the proportion of patients with hospital-acquired VTE that are preventable. Methods: This was a retrospective study of patients in two tertiary care hospitals in Sydney, Australia. Data were collected for patients with hospital-acquired VTE based on International Statistical Classification of Diseases and Related Health Problems, 10th Revision, Australian Modification (ICD-10-AM) coding from January 2018 to May 2020. Patients were classified as low, moderate or high risk of developing a VTE during hospitalisation based on demographic and clinical factors. A hospital-acquired VTE was considered to be potentially preventable if there was suboptimal prophylaxis in the absence of contraindications. Suboptimal therapy included at least one of the following related to VTE prophylaxis: low dose, missed dose (prior to developing a VTE), suboptimal drug and delayed start (>24 h from admission). Results: There were 229 patients identified with VTE based on ICD-10-AM coding. A subset of 135 patients were determined to have actual hospital-acquired VTE. Of these, there were no patients at low risk, 64% (87/135) at moderate risk and 44% (48/135) at high risk of developing a VTE. Most (65%; n = 88/135) patients had one or more contraindications to receive recommended prophylaxis. Overall, the proportion of patients who received suboptimal prophylaxis was 11% (15/135). Conclusion: Approximately one out of 10 hospital-acquired VTE are preventable. Hospitals should focus on measuring and reporting VTE that are preventable to provide a more accurate measure of the burden of VTE that can be reduced by improving care.
MeSH term(s)	Humans ; Venous Thromboembolism/epidemiology ; Venous Thromboembolism/prevention & control ; Venous Thromboembolism/drug therapy ; Retrospective Studies ; Australia/epidemiology ; Hospitalization ; Hospitals ; Risk Factors ; Anticoagulants/therapeutic use
Chemical Substances	Anticoagulants
Language	English
Publishing date	2022-08-13
Publishing country	Australia
Document type	Journal Article
ZDB-ID	2045436-3
ISSN	1445-5994 ; 1444-0903
ISSN (online)	1445-5994
ISSN	1444-0903
DOI	10.1111/imj.15600
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Zs.A 792: Show issues			Location: Je nach Verfügbarkeit (siehe Angabe bei Bestand) bis Jg. 1994: Bestellungen von Artikeln über das Online-Bestellformular Jg. 1995 - 2021: Lesesall (1.OG) ab Jg. 2022: Lesesaal (EG)
Zs.MO 424: Show issues

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Article ; Online: Emicizumab assay evaluations and results from an Australian field study of emicizumab measurement.

Kershaw, Geoffrey / Dix, Caroline / Chen, Vivien M / Cai, Nancy / Khoo, Teh-Liane

Pathology

2022 Volume 54, Issue 6, Page(s) 755–762

Abstract: Emicizumab is a recombinant, humanised bispecific antibody which acts as a FVIII mimetic and is a therapeutic option for haemophilia A. Plasma emicizumab levels may sometimes be required. Multiple one-stage clotting assays (OSA) and one human component ... ...

Abstract	Emicizumab is a recombinant, humanised bispecific antibody which acts as a FVIII mimetic and is a therapeutic option for haemophilia A. Plasma emicizumab levels may sometimes be required. Multiple one-stage clotting assays (OSA) and one human component chromogenic assay (CSA) were used to measure emicizumab both centrally and by a field study. The study samples drug concentrations range included within therapy range of 35-70 μg/mL. All assays were modified from traditional FVIII assays to enable replacement of plasma calibrators with emicizumab calibrators. Central laboratory OSA mean recovery levels (target) for six spike levels of emicizumab were close to target at 120.5 (120), 81.6 (80), 40.9 (40), 21.4 (20), 10.7 (10) and 5.5 (5) μg/mL. Field study OSA mean recoveries were similarly close to target. Between method coefficients of variation were <9% in both the central laboratory and field study assays, except for the 5 μg/mL sample which was 12.3%. CSA mean recoveries were within 10% of target at 80, 50 and 20 μg/mL levels. This study affirms that emicizumab can be measured by OSA using many types of activated partial thromboplastin time (APTT) reagents and is also measurable by the human CSA. The assays showed good precision, accuracy and linearity both locally and in a field study setting.
MeSH term(s)	Humans ; Antibodies, Bispecific/pharmacology ; Antibodies, Bispecific/therapeutic use ; Antibodies, Monoclonal, Humanized ; Australia ; Factor VIII/therapeutic use ; Hemophilia A/diagnosis ; Hemophilia A/drug therapy
Chemical Substances	Antibodies, Bispecific ; Antibodies, Monoclonal, Humanized ; emicizumab (7NL2E3F6K3) ; Factor VIII (9001-27-8)
Language	English
Publishing date	2022-05-14
Publishing country	England
Document type	Journal Article
ZDB-ID	7085-3
ISSN	1465-3931 ; 0031-3025
ISSN (online)	1465-3931
ISSN	0031-3025
DOI	10.1016/j.pathol.2022.02.006
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Article ; Online: Colchicine as a Modulator of Platelet Function: A Systematic Review.

Reddel, Caroline J / Pennings, Gabrielle J / Chen, Vivien M / Gnanenthiran, Sonali / Kritharides, Leonard

Seminars in thrombosis and hemostasis

2022 Volume 48, Issue 5, Page(s) 552–567

Abstract: The microtubule inhibitor and anti-inflammatory agent colchicine is used to treat a range of conditions involving inflammasome activation in monocytes and neutrophils, and is now known to prevent coronary and cerebrovascular events. In vitro studies ... ...

Abstract	The microtubule inhibitor and anti-inflammatory agent colchicine is used to treat a range of conditions involving inflammasome activation in monocytes and neutrophils, and is now known to prevent coronary and cerebrovascular events. In vitro studies dating back more than 50 years showed a direct effect of colchicine on platelets, but as little contemporary attention has been paid to this area, we have critically reviewed the effects of colchicine on diverse aspects of platelet biology in vitro and in vivo. In this systematic review we searched Embase, Medline, and PubMed for articles testing platelets after incubation with colchicine and/or reporting a clinical effect of colchicine treatment on platelet function, including only papers available in English and excluding reviews and conference abstracts. We identified 98 relevant articles and grouped their findings based on the type of study and platelet function test. In vitro, colchicine inhibits traditional platelet functions, including aggregation, clotting, degranulation, and platelet-derived extracellular vesicle formation, although many of these effects were reported at apparently supraphysiological concentrations. Physiological concentrations of colchicine inhibit collagen- and calcium ionophore-induced platelet aggregation and internal signaling. There have been limited studies of in vivo effects on platelets. The colchicine-platelet interaction has the potential to contribute to colchicine-mediated reduction in cardiovascular events, but there is a pressing need for high quality clinical research in this area.
MeSH term(s)	Blood Platelets ; Colchicine/pharmacology ; Colchicine/therapeutic use ; Hemostasis ; Humans ; Platelet Aggregation ; Platelet Function Tests
Chemical Substances	Colchicine (SML2Y3J35T)
Language	English
Publishing date	2022-07-26
Publishing country	United States
Document type	Journal Article ; Systematic Review
ZDB-ID	196901-8
ISSN	1098-9064 ; 0094-6176
ISSN (online)	1098-9064
ISSN	0094-6176
DOI	10.1055/s-0042-1749660
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Article ; Online: Tinzaparin for venous thromboembolism in patients with renal impairment: a single-centre, prospective pilot study.

Yeung, James / Dix, Caroline H K / Ritchie, Angus G / Kow, Marian / Chen, Vivien M Y

Internal medicine journal

2022 Volume 53, Issue 1, Page(s) 68–73

Abstract: Background: Low molecular weight heparins (LMWH) are used extensively for prophylaxis and treatment of venous thromboembolism (VTE), bridging therapy for warfarin and standard of care in cancer-associated VTE (CA-VTE). Tinzaparin has the highest ... ...

Abstract	Background: Low molecular weight heparins (LMWH) are used extensively for prophylaxis and treatment of venous thromboembolism (VTE), bridging therapy for warfarin and standard of care in cancer-associated VTE (CA-VTE). Tinzaparin has the highest molecular weight of all LMWH and relies least on renal clearance to Cockcroft-Gault creatinine clearance (CrCl) of 20 mL/min. Previous pharmacological studies have demonstrated safety and effectiveness in elderly patients. Prospective clinical trials have confirmed these findings to CrCl 20 mL/min and in CA-VTE. We describe the pilot program developed at Concord Repatriation General Hospital for tinzaparin. Aims: We aim to confirm the deliverability of tinzaparin in patients with renal insufficiency. Methods: Twenty patients were established on tinzaparin as therapeutic anticoagulation with CrCl or CKD-EPI estimated glomerular filtration rate (eGFR) 20-50 mL/min with an indication for anticoagulation. Tinzaparin was given as a subcutaneous injection at 175 units/kg as a single daily dose, rounded to the nearest vial size. Tinzaparin anti-Xa levels were tested at Days 2, 7 and 14 (±1 day) and transition to oral anticoagulants were allowed at clinician discretion. Results: No accumulation of tinzaparin was seen into Day 14. Two patients required dose-adjustment, five patients had bleeding complications (two major, three minor) and four patients died during follow-up, all attributable to patients' comorbidities. CrCl and body surface area-standardised CrCl were significantly correlated with tinzaparin anti-Xa level only on Day 2, and this effect was lost when patients with CrCl >50 mL/min were excluded. Data from our cohort confirm the deliverability of therapeutic tinzaparin in patients with CrCl or CKD-EPI eGFR 20-50 mL/min. Bleeding and death outcomes were also comparable to other trials using tinzaparin in CA-VTE. Conclusion: For patients with renal insufficiency, tinzaparin represents an attractive alternative anticoagulant with once-daily administration in a range of potential indications.
MeSH term(s)	Humans ; Aged ; Tinzaparin/therapeutic use ; Heparin, Low-Molecular-Weight/therapeutic use ; Pilot Projects ; Venous Thromboembolism/prevention & control ; Prospective Studies ; Anticoagulants/adverse effects ; Renal Insufficiency/chemically induced ; Renal Insufficiency, Chronic/drug therapy
Chemical Substances	Tinzaparin (7UQ7X4Y489) ; Heparin, Low-Molecular-Weight ; Anticoagulants
Language	English
Publishing date	2022-12-05
Publishing country	Australia
Document type	Journal Article
ZDB-ID	2045436-3
ISSN	1445-5994 ; 1444-0903
ISSN (online)	1445-5994
ISSN	1444-0903
DOI	10.1111/imj.15010
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Zs.A 792: Show issues			Location: Je nach Verfügbarkeit (siehe Angabe bei Bestand) bis Jg. 1994: Bestellungen von Artikeln über das Online-Bestellformular Jg. 1995 - 2021: Lesesall (1.OG) ab Jg. 2022: Lesesaal (EG)
Zs.MO 424: Show issues

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Article ; Online: In vitro

Lee, Christine S M / Huguenin, Yoann / Pillois, Xavier / Moulieras, Mikeldi / Marcy, Ella / Whittaker, Shane / Chen, Vivien M Y / Fiore, Mathieu

Research and practice in thrombosis and haemostasis

2023 Volume 8, Issue 1, Page(s) 102253

Abstract: Background: Glanzmann thrombasthenia (GT) is a rare bleeding disorder caused by inherited defects of the platelet α: Objectives: In this study, we aimed to determine the prevalence of such isoantibodies and better characterize their pathogenic ... ...

Abstract	Background: Glanzmann thrombasthenia (GT) is a rare bleeding disorder caused by inherited defects of the platelet α Objectives: In this study, we aimed to determine the prevalence of such isoantibodies and better characterize their pathogenic properties. Methods: Twelve patients with GT were evaluated for anti-α Results: Only 2 samples were able to severely block integrin function. We observed that these 2 sera caused a reduction in platelet size similar to that observed when platelets become procoagulant. Mixing healthy donor platelets with patients' sera or purified IgGs led to microvesiculation, phosphatidylserine exposure, and induction of calcium influx. This was associated with an increase in procoagulant platelets. Pore formation and calcium entry were associated with complement activation, leading to the constitution of a membrane attack complex (MAC) with enhanced complement protein C5b-9 formation. This process was inhibited by the complement 5 inhibitor eculizumab and reduced by polyvalent human immunoglobulins. Conclusion: Our data suggest that complement activation induced by rare blocking anti-α
Language	English
Publishing date	2023-11-04
Publishing country	United States
Document type	Journal Article
ISSN	2475-0379
ISSN (online)	2475-0379
DOI	10.1016/j.rpth.2023.102253
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Article ; Online: Platelets from patients with myeloproliferative neoplasms have increased numbers of mitochondria that are hypersensitive to depolarization by thrombin.

Ross, David M / Liang, Hai Po Helena / Iqra, Zeenet / Whittaker, Shane / Tan, Chuen Wen / Dale, Brian J / Chen, Vivien M

Scientific reports

2023 Volume 13, Issue 1, Page(s) 9172

Abstract: Thrombosis is one of the cardinal manifestations of myeloproliferative neoplasms (MPN). The mechanisms leading to a prothrombotic state in MPN are complex and remain poorly understood. Platelet mitochondria play a role in platelet activation, but their ... ...

Abstract	Thrombosis is one of the cardinal manifestations of myeloproliferative neoplasms (MPN). The mechanisms leading to a prothrombotic state in MPN are complex and remain poorly understood. Platelet mitochondria play a role in platelet activation, but their number and function have not been extensively explored in MPN to date. We observed an increased number of mitochondria in platelets from MPN patients compared with healthy donors. MPN patients had an increased proportion of dysfunctional platelet mitochondria. The fraction of platelets with depolarized mitochondria at rest was increased in essential thrombocythemia (ET) patients and the mitochondria were hypersensitive to depolarization following thrombin agonist stimulation. Live microscopy showed a stochastic process in which a higher proportion of individual ET platelets underwent mitochondrial depolarization and after a shorter agonist exposure compared to healthy donors. Depolarization was immediately followed by ballooning of the platelet membrane, which is a feature of procoagulant platelets. We also noted that the mitochondria of MPN patients were on average located nearer the platelet surface and we observed extrusion of mitochondria from the platelet surface as microparticles. These data implicate platelet mitochondria in a number of prothrombotic phenomena. Further studies are warranted to assess whether these findings correlate with clinical thrombotic events.
MeSH term(s)	Humans ; Blood Platelets/metabolism ; Thrombin/metabolism ; Myeloproliferative Disorders/metabolism ; Thrombocythemia, Essential/metabolism ; Thrombosis/metabolism ; Platelet Activation ; Neoplasms/metabolism ; Mitochondria
Chemical Substances	Thrombin (EC 3.4.21.5)
Language	English
Publishing date	2023-06-06
Publishing country	England
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2615211-3
ISSN	2045-2322 ; 2045-2322
ISSN (online)	2045-2322
ISSN	2045-2322
DOI	10.1038/s41598-023-36266-2
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