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Effects of emicizumab (0.15 mg/mL) on the clot waveform analysis (CWA). (a) Activated partial thromboplastin time (APTT). (b) thrombin time (TT); upper, normal plasma (NP); lower, FVIII-deficient plasma (FVIIID); Emi, emicizumab; navy-blue, fibrin formation curve; pink curve, 1st derivative curve; light-blue curve, 2nd derivative curve. All assays were performed more than three times to confirm the reproducibility.
Source publication
Objective:
Although emicizumab is a bispecific, monoclonal antibody that has led to a significant improvement of treatment for hemophilia A patients with inhibitors, the routine monitoring of patients treated with emicizumab is difficult. Thrombin time (TT) reflects thrombin burst, which mainly depends on activation of factor V (FV) and FVIII.
Me...
Contexts in source publication
Context 1
... (0.15 mg/mL) caused clot formation in FVIII-deficient plasma, causing the peak of CWA-APTT to appear, whereas it shortened the peak time of the CWA-APTT in normal plasma but did not increase the peak height of the CWA-APTT (Figure 1a). Regarding the sTF/FIXa assay, emicizumab slightly decreased the peak heights and enlarged the peak width in both normal and FVIII-deficient plasma (Supplementary Figure S1). ...
Context 2
... (0.15 mg/mL) caused clot formation in FVIII-deficient plasma, causing the peak of CWA-APTT to appear, whereas it shortened the peak time of the CWA-APTT in normal plasma but did not increase the peak height of the CWA-APTT (Figure 1a). Regarding the sTF/FIXa assay, emicizumab slightly decreased the peak heights and enlarged the peak width in both normal and FVIII-deficient plasma (Supplementary Figure S1). The peak height of the FF curve was significantly higher in normal plasma than in FVIII-deficient plasma. ...
Context 3
... peak height of the FF curve was significantly higher in normal plasma than in FVIII-deficient plasma. Regarding the CWA-TT using 0.5 IU/mL of thrombin, the peak times and heights of CWA-TT in both normal and FVIII-deficient plasma were similar between samples with and without emicizumab, whereas the peak heights of the FF curve with and without emicizumab were markedly higher (p < 0.001) in normal plasma (1366 ± 67 mm absorbance and 1380 ±56 mm absorbance, respectively) than in FVIII-deficient plasma (771 ± 54 mm absorbance and 758 ± 37 mm absorbance, respectively) ( Figure 1b). ...
Context 4
... clinical trials for emicizumab (HAVEN 1-4) in hemophilia A patients with or without inhibitors showed the significant improvement of treatments for hemophilia [6,7,24,25]. Although HAVEN 1 showed several thrombotic complications in combination with activated plasma prothrombin complex reagents [6], there were further new thrombotic complications in the long-term outcomes in the HAVEN 1-4 studies [26]. ...
Context 5
... Materials: The following supporting information can be downloaded at: https://www.mdpi.com/article/10.3390/jcm11206142/s1, Figure S1: Effects of emicizumab (0.15 mg/mL) on the clot waveform analysis (CWA)-small amount of tissue factor-induced FIX activation assay (sTF/FIXa); Figure S2: Effects of emicizumab on the clot waveform analysis (CWA)-thrombin time (TT); Figure S3: Effects of emicizumab on mixing test using the clot waveform analysis (CWA) -activated partial thromboplastin time (APTT) (a) and CWA-thrombin time (TT) (b). Upper, Emi (+); Lower, Emi (-) NP, normal plasma; FVIIID, FVIII-deficient plasma; Emi (+), with emicizumab; Emi (-), without emicizumab; navy blue, fibrin formation curve; pink curve, 1 st derivative curve; light blue curve, 2 nd derivative curve. ...
Similar publications
Congenital factor V (FV) and factor VII (FVII) deficiency is a rare coagulopathy, caused by two distinct independently segregating genetic defects. A 54-year-old female presented to hospital with non-ST elevation myo-cardial infarction. Workup revealed an elevated international normalized ratio (INR) of 2.0 (normal range 0.8–1.2). Thrombin time (TT...
Citations
... Apart from APTT, TT reflects the time required for thrombin to convert fibrinogen to fibrin. FIB is the substrate of the process, so its level affects the length of TT. 30,31 Based on the activation of FVIII, TT can reflect a thrombin burst, which was synergistic with the generation of FIXa caused by the process that APTT activated the intrinsic pathway, thus contributing to the conversion of FIB to fibrin. 31,32 In our study, the results suggested the decrease of TT with accompanying by the increase of FIB. ...
... FIB is the substrate of the process, so its level affects the length of TT. 30,31 Based on the activation of FVIII, TT can reflect a thrombin burst, which was synergistic with the generation of FIXa caused by the process that APTT activated the intrinsic pathway, thus contributing to the conversion of FIB to fibrin. 31,32 In our study, the results suggested the decrease of TT with accompanying by the increase of FIB. The shorter TT in the adenomyosis patients than in the patients with uterine leiomyoma or the control group may be due to the higher level of FIB in the adenomyosis. ...
Objective
Adenomyosis patients are in a hypercoagulable state, and studies have shown that carbohydrate antigen125 (CA125) may relate to the hypercoagulability and thrombosis of patients with adenomyosis, but there is still a lack of clarity regarding the changes in CA125-related coagulation indicators. This study was to explore the changes and influencing factors of CA125-related coagulation parameters in patients with adenomyosis.
Methods
Retrospective observational study conducted on 200 patients with adenomyosis (AM group), 240 patients with uterine leiomyoma (LM group) and 81 patients with cervical intraepithelial neoplasia (CIN)-III (control group), of which the coagulation parameters were detected by clinical blood sample collection and statistical method analysis and informed consent was obtained.
Results
The level of CA125 in the AM group was significantly higher than that in the LM group and control group. However, thrombin time (TT) shortened in the AM group when compared with the LM and control group. Activated partial thromboplastin time (APTT) in the AM group was shorter than in the control group. Multivariate logistic regression analysis found that adenomyosis was associated with CA125 level (OR=323.860, 95% CI 90.424–1159.924, P<0.001), APTT (OR=1.295, 95% CI 1.050–1.598, P=0.016), TT (OR=0.642, 95% CI 0.439–0.938, P=0.022), menorrhagia (OR=7.363, 95% CI 2.544–21.315, P<0.001), dysmenorrhea (OR=22.590, 95% CI 8.185–62.347, P<0.001). Correlation analysis revealed that APTT (r= −0.207) and TT (r = −0.174) were negatively correlated with the level of CA125.
Conclusion
The shortening of CA125-related APTT and TT indicates that it is meaningful to detect coagulation parameters of patients with elevated CA125 levels early, dysmenorrhea and menorrhagia, and maybe further discover the hypercoagulability and prevent the occurrence of thrombus in adenomyosis.
... A small amount of thrombin reflects the thrombin burst, including the activation of FXI, FVIII, and FV instead of fibrinogen, and thrombin time (TT) using only a small amount of thrombin can evaluate the intrinsic pathway and FVIII activity [16]. Recently, it was reported that CWA-TT can measure FVIII activity independent of the presence of emicizumab [17]. ...
... This study was conducted in accordance with the principles of the Declaration of Helsinki. The CWA-TT ( Figure 1a) using 0.5 IU thrombin (Thrombin 500 units; Mochida Pharmaceutical Co., Ltd., Tokyo, Japan) was measured using an ACL-TOP ® system (Instrumentation Laboratory, Bedford, MA, USA) [16,17]. This system shows three types of curves [16,17]. ...
... The CWA-TT ( Figure 1a) using 0.5 IU thrombin (Thrombin 500 units; Mochida Pharmaceutical Co., Ltd., Tokyo, Japan) was measured using an ACL-TOP ® system (Instrumentation Laboratory, Bedford, MA, USA) [16,17]. This system shows three types of curves [16,17]. One illustrates the changes in the absorbance observed while measuring CWA-TT, corresponding to the fibrin formation curve (FFC). ...
Background:
Regular prophylactic therapy has become an increasingly common treatment for severe hemophilia. Therefore, hypercoagulability-a potential risk factor of thrombosis-is a cause for concern in hemophilic patients treated with a high dose of FVIII concentrate. In clot waveform analysis (CWA)-thrombin time (TT), a small amount of thrombin activates clotting factor VIII (FVIII) instead of fibrinogen, resulting in FVIII measurements using CWA-TT with a small amount of thrombin.
Methods:
The coagulation ability of patients treated with FVIII concentrate or emicizumab was evaluated using activated partial thromboplastin time (APTT), TT and a small amount of tissue factor-induced FIX activation assay (sTF/FIXa) using CWA.
Results:
The FVIII activity based on CWA-TT was significantly greater than that based on the CWA-APTT or chromogenic assay. FVIII or FVIII-like activities based on the three assays in plasma without emicizumab were closely correlated; those in plasma with emicizumab based on CWA-TT and chromogenic assays were also closely correlated. CWA-APTT and CWA-TT showed different patterns in patients treated with FVIII concentrates compared to those treated with emicizumab. In particular, CWA-TT in patients treated with FVIII concentrate showed markedly higher peaks in platelet-rich plasma than in platelet-poor plasma. CWA-APTT showed lower coagulability in hemophilic patients treated with FVIII concentrate than in healthy volunteers, whereas CWA-sTF/FIXa did not. In contrast, CWA-TT showed hypercoagulability in hemophilic patients treated with FVIII concentrate.
Conclusions:
CWA-TT can be used to evaluate the thrombin bursts that cause hypercoagulability in patients treated with emicizumab. Although routine APTT evaluations demonstrated low coagulation ability in patients treated with FVIII concentrate, CWA-TT showed hypercoagulability in these patients, suggesting that the evaluation of coagulation ability may be useful when using multiple assays.
... However, few reports have described this relationship between the FVIII activity assessed using the peak time and height of CWA-APTT, including a small amount of tissue factor-induced activated FIX (sTF/FIXa) assay [14]. A CWA-small amount of thrombin time (CWA-TT) also reflects thrombin burst and FVIII activity [15] and can be used to measure the FVIII activity independent of the presence of emicizumab [16]. ...
... This study was carried out in accordance with the principles of the Declaration of Helsinki. The CWA-TT was measured using 0.5 IU thrombin (Thrombin 500 units; Mochida Pharmaceutical Co., Ltd., Tokyo, Japan) with an ACL-TOP ® system (Instrumentation Laboratory, Bedford, MA, USA) [15,16]. Three types of curves are shown on this system monitor [15,16]. ...
... The CWA-TT was measured using 0.5 IU thrombin (Thrombin 500 units; Mochida Pharmaceutical Co., Ltd., Tokyo, Japan) with an ACL-TOP ® system (Instrumentation Laboratory, Bedford, MA, USA) [15,16]. Three types of curves are shown on this system monitor [15,16]. One shows the changes in the absorbance observed while measuring the TT, corresponding to the fibrin formation curve (FFC). ...
Background: Although the use of regular replacement therapy including emicizumab for severe hemophilia has been spread, the assessment of the hemostatic ability using routine activated partial thromboplastin time (APTT) is still difficult in patients being treated with emicizumab. Methods: The hemostatic ability in patients treated with FVIII concentrate or emicizumab was evaluated by APTT, thrombin time (TT) and a small amount of tissue factor induced FIX activation assay (sTF/FIXa) using a clot waveform analysis (CWA). Results: FVIII activities based on a CWA-TT were significantly higher than those based on a CWA-APTT or chromogenic assay. FVIII activities based on the three assays in plasma without emicizumab were closely correlated, and those in plasma with emicizumab based on a CWA-TT and chromogenic assays were also closely correlated. The CWA-APTT and CWA-TT showed different patterns in patients treated with FVIII concentrates from those treated with emicizumab. In particular, the CWA-TT in patients treated with FVIII concentrate showed that the peak heights were significantly higher in platelet-rich plasma than in platelet-poor plasma. In plasma with approximately 16% of FVIII activity based on APTT assay from patients treated with FVIII concentrate, the peak height on the CWA-sTF/FIXa showed a higher hemostatic abilitythan normal plasma. Conclusions: The CWA-TT can measure the FVIII activity in patients treated with emicizumab. Although routine APTT evaluations demonstrate a low hemostatic ability in patients treated with FVIII concentrate, the CWA-TT and CWA-sTF/FIXa show hypercoagulability in those patients.
... The measurement of FVIII activity by APTT is difficult in patients with hemophilia A treated with emicizumab [40]; however, CWA-dilute TT can measure FVIII activity in plasma independent of the presence of emicizumab [41] (Fig. 6). Therefore, CWA-dilute TT may be useful for monitoring hemostasis in patients with hemophilia treated with emicizumab (Fig. 6). ...
Clot waveform analysis (CWA) observes changes in transparency in a plasma sample based on clotting tests such as activated partial thromboplastin time (APTT), prothrombin time (PT), and thrombin time (TT). Evidence indicates that not only an abnormal waveform but also peak times and heights in derivative curves of CWA are useful for the evaluation of hemostatic abnormalities. Modified CWA, including the PT with APTT reagent, dilute PT (small amount of tissue factor [TF]-induced clotting factor IX [FIX] activation; sTF/FIXa), and dilute TT, has been proposed to evaluate physiological or pathological hemostasis. We review routine and modified CWA and their clinical applications. In CWA-sTF/FIXa, elevated peak heights indicate hypercoagulability in patients with cancer or thrombosis, whereas prolonged peak times indicate hypocoagulability in several conditions, including clotting factor deficiency and thrombocytopenia. CWA-dilute TT reflects the thrombin burst, whereas clot-fibrinolysis waveform analysis reflects both hemostasis and fibrinolysis. The relevance and usefulness of CWA-APTT and modified CWA should be further investigated in various diseases.
