Alterations and impact of thrombin generation and clot formation in solvent/detergent plasma, FXIII deficiency and lysinuric protein intolerance

Hanna Pitkänen

Research output: ThesisDoctoral ThesisCollection of Articles

Abstract

Hemostasis is a crucial biological process, in which thrombin generation (TG) and formation of fibrin network are key elements. Equally important is the capability to dissolve the blood clots through fibrinolysis to repair blood flow. This thesis seeks to recognize factors influencing TG, fibrin formation and fibrinolysis related to solvent/detergent (S/D) plasma, congenital factor XIII (FXIII) deficiency and lysinuric protein intolerance (LPI). In the first in vitro study, several human plasma preparations were used to elucidate the impact of the SD method on TG and clot stability. In the second study FXIII concentrate therapies were monitored in congenital FXIII deficiency. In the third study the hemostatic and fibrinolytic capacity of 15 LPI patients was investigated. Both traditional and functional coagulation assays were performed. GPRP-peptide was used to inhibit fibrinogen polymerization. Optical density derivatives were assessed to monitor fibrin formation and degradation, and fibrinolysis markers were determined. Traditional coagulation markers measure the time from the exogenous activation of coagulation to the initiation of clot formation, or the levels of coagulation factors in plasma. Functional coagulation assays can display the concentration of thrombin in plasma or provide information of clot formation kinetics, stability, or fibrinolysis. By combining traditional coagulation screening with the functional assays, our study revealed increased fibrinolysis in SD plasma, while augmented fibrinolysis could be excluded, even at trough FXIII levels in congenital FXIII deficiency. In LPI derivatives of fibrin formation and fibrinolysis confirmed their abnormalities, also detected by in vivo markers. Enhanced TG and fibrinolysis were observed in SD plasma. SD treatment reduced the levels of single chain protein S, leading to a loss of both activated protein C -dependent and independent protein S activity, enhancing TG. In addition, the in vitro coagulation phenotype of SD plasma was abnormal due to increased fibrinolysis likely via diminished levels of a2-antiplasmin (a2-AP). Tranexamic acid abolished this fibrinolysis in vitro. Low levels of FXIII induced accelerated prothrombin conversion, leading to increased TG, possibly based on decreased fibrin antithrombin I (AT I) -like activity. According to AT I theory forming fibrin (= AT I) inhibits TG during coagulation process by sequestering thrombin. In our study, impaired fibrin formation induced both by lack of FXIII and GPRP administration in normal plasma enhanced TG, supporting the AT I hypothesis. In LPI, a metabolic disorder leading to renal insufficiency, we discovered impaired primary hemostasis and altered TG. Moreover, we detected decreased fibrin formation and markedly enhanced fibrinolysis that both related to altered plasminogen/a2-AP ratio and renal insufficiency. Platelet levels, function of primary hemostasis, fibrinogen and FXIII levels should be measured perioperatively and treated accordingly with platelet transfusions and fibrinogen and FXIII concentrates (in LPI). In summary, combining traditional and functional coagulation assays is beneficial. The altered coagulation phenotype of SD plasma should be acknowledged during transfusion therapy. Enhanced TG under low FXIII levels may guide FXIII replacement therapy. Our results concerning LPI further confirm the link between impaired coagulation and renal insufficiency.
Original languageEnglish
Supervisors/Advisors
  • Lassila, Riitta , Supervisor
Award date23 Nov 2018
Place of PublicationHelsinki
Publisher
Print ISBNs978-951-51-4560-4
Electronic ISBNs978-951-51-4561-1
Publication statusPublished - 2018
MoE publication typeG5 Doctoral dissertation (article)

Fields of Science

  • 3121 Internal medicine

Cite this

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title = "Alterations and impact of thrombin generation and clot formation in solvent/detergent plasma, FXIII deficiency and lysinuric protein intolerance",
abstract = "Hemostasis is a crucial biological process, in which thrombin generation (TG) and formation of fibrin network are key elements. Equally important is the capability to dissolve the blood clots through fibrinolysis to repair blood flow. This thesis seeks to recognize factors influencing TG, fibrin formation and fibrinolysis related to solvent/detergent (S/D) plasma, congenital factor XIII (FXIII) deficiency and lysinuric protein intolerance (LPI). In the first in vitro study, several human plasma preparations were used to elucidate the impact of the SD method on TG and clot stability. In the second study FXIII concentrate therapies were monitored in congenital FXIII deficiency. In the third study the hemostatic and fibrinolytic capacity of 15 LPI patients was investigated. Both traditional and functional coagulation assays were performed. GPRP-peptide was used to inhibit fibrinogen polymerization. Optical density derivatives were assessed to monitor fibrin formation and degradation, and fibrinolysis markers were determined. Traditional coagulation markers measure the time from the exogenous activation of coagulation to the initiation of clot formation, or the levels of coagulation factors in plasma. Functional coagulation assays can display the concentration of thrombin in plasma or provide information of clot formation kinetics, stability, or fibrinolysis. By combining traditional coagulation screening with the functional assays, our study revealed increased fibrinolysis in SD plasma, while augmented fibrinolysis could be excluded, even at trough FXIII levels in congenital FXIII deficiency. In LPI derivatives of fibrin formation and fibrinolysis confirmed their abnormalities, also detected by in vivo markers. Enhanced TG and fibrinolysis were observed in SD plasma. SD treatment reduced the levels of single chain protein S, leading to a loss of both activated protein C -dependent and independent protein S activity, enhancing TG. In addition, the in vitro coagulation phenotype of SD plasma was abnormal due to increased fibrinolysis likely via diminished levels of a2-antiplasmin (a2-AP). Tranexamic acid abolished this fibrinolysis in vitro. Low levels of FXIII induced accelerated prothrombin conversion, leading to increased TG, possibly based on decreased fibrin antithrombin I (AT I) -like activity. According to AT I theory forming fibrin (= AT I) inhibits TG during coagulation process by sequestering thrombin. In our study, impaired fibrin formation induced both by lack of FXIII and GPRP administration in normal plasma enhanced TG, supporting the AT I hypothesis. In LPI, a metabolic disorder leading to renal insufficiency, we discovered impaired primary hemostasis and altered TG. Moreover, we detected decreased fibrin formation and markedly enhanced fibrinolysis that both related to altered plasminogen/a2-AP ratio and renal insufficiency. Platelet levels, function of primary hemostasis, fibrinogen and FXIII levels should be measured perioperatively and treated accordingly with platelet transfusions and fibrinogen and FXIII concentrates (in LPI). In summary, combining traditional and functional coagulation assays is beneficial. The altered coagulation phenotype of SD plasma should be acknowledged during transfusion therapy. Enhanced TG under low FXIII levels may guide FXIII replacement therapy. Our results concerning LPI further confirm the link between impaired coagulation and renal insufficiency.",
keywords = "3121 Internal medicine",
author = "Hanna Pitk{\"a}nen",
note = "M1 - 81 s. + liitteet",
year = "2018",
language = "English",
isbn = "978-951-51-4560-4",
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number = "68/2018",
address = "Finland",

}

Alterations and impact of thrombin generation and clot formation in solvent/detergent plasma, FXIII deficiency and lysinuric protein intolerance. / Pitkänen, Hanna.

Helsinki : Helsingin yliopisto, 2018. 81 p.

Research output: ThesisDoctoral ThesisCollection of Articles

TY - THES

T1 - Alterations and impact of thrombin generation and clot formation in solvent/detergent plasma, FXIII deficiency and lysinuric protein intolerance

AU - Pitkänen, Hanna

N1 - M1 - 81 s. + liitteet

PY - 2018

Y1 - 2018

N2 - Hemostasis is a crucial biological process, in which thrombin generation (TG) and formation of fibrin network are key elements. Equally important is the capability to dissolve the blood clots through fibrinolysis to repair blood flow. This thesis seeks to recognize factors influencing TG, fibrin formation and fibrinolysis related to solvent/detergent (S/D) plasma, congenital factor XIII (FXIII) deficiency and lysinuric protein intolerance (LPI). In the first in vitro study, several human plasma preparations were used to elucidate the impact of the SD method on TG and clot stability. In the second study FXIII concentrate therapies were monitored in congenital FXIII deficiency. In the third study the hemostatic and fibrinolytic capacity of 15 LPI patients was investigated. Both traditional and functional coagulation assays were performed. GPRP-peptide was used to inhibit fibrinogen polymerization. Optical density derivatives were assessed to monitor fibrin formation and degradation, and fibrinolysis markers were determined. Traditional coagulation markers measure the time from the exogenous activation of coagulation to the initiation of clot formation, or the levels of coagulation factors in plasma. Functional coagulation assays can display the concentration of thrombin in plasma or provide information of clot formation kinetics, stability, or fibrinolysis. By combining traditional coagulation screening with the functional assays, our study revealed increased fibrinolysis in SD plasma, while augmented fibrinolysis could be excluded, even at trough FXIII levels in congenital FXIII deficiency. In LPI derivatives of fibrin formation and fibrinolysis confirmed their abnormalities, also detected by in vivo markers. Enhanced TG and fibrinolysis were observed in SD plasma. SD treatment reduced the levels of single chain protein S, leading to a loss of both activated protein C -dependent and independent protein S activity, enhancing TG. In addition, the in vitro coagulation phenotype of SD plasma was abnormal due to increased fibrinolysis likely via diminished levels of a2-antiplasmin (a2-AP). Tranexamic acid abolished this fibrinolysis in vitro. Low levels of FXIII induced accelerated prothrombin conversion, leading to increased TG, possibly based on decreased fibrin antithrombin I (AT I) -like activity. According to AT I theory forming fibrin (= AT I) inhibits TG during coagulation process by sequestering thrombin. In our study, impaired fibrin formation induced both by lack of FXIII and GPRP administration in normal plasma enhanced TG, supporting the AT I hypothesis. In LPI, a metabolic disorder leading to renal insufficiency, we discovered impaired primary hemostasis and altered TG. Moreover, we detected decreased fibrin formation and markedly enhanced fibrinolysis that both related to altered plasminogen/a2-AP ratio and renal insufficiency. Platelet levels, function of primary hemostasis, fibrinogen and FXIII levels should be measured perioperatively and treated accordingly with platelet transfusions and fibrinogen and FXIII concentrates (in LPI). In summary, combining traditional and functional coagulation assays is beneficial. The altered coagulation phenotype of SD plasma should be acknowledged during transfusion therapy. Enhanced TG under low FXIII levels may guide FXIII replacement therapy. Our results concerning LPI further confirm the link between impaired coagulation and renal insufficiency.

AB - Hemostasis is a crucial biological process, in which thrombin generation (TG) and formation of fibrin network are key elements. Equally important is the capability to dissolve the blood clots through fibrinolysis to repair blood flow. This thesis seeks to recognize factors influencing TG, fibrin formation and fibrinolysis related to solvent/detergent (S/D) plasma, congenital factor XIII (FXIII) deficiency and lysinuric protein intolerance (LPI). In the first in vitro study, several human plasma preparations were used to elucidate the impact of the SD method on TG and clot stability. In the second study FXIII concentrate therapies were monitored in congenital FXIII deficiency. In the third study the hemostatic and fibrinolytic capacity of 15 LPI patients was investigated. Both traditional and functional coagulation assays were performed. GPRP-peptide was used to inhibit fibrinogen polymerization. Optical density derivatives were assessed to monitor fibrin formation and degradation, and fibrinolysis markers were determined. Traditional coagulation markers measure the time from the exogenous activation of coagulation to the initiation of clot formation, or the levels of coagulation factors in plasma. Functional coagulation assays can display the concentration of thrombin in plasma or provide information of clot formation kinetics, stability, or fibrinolysis. By combining traditional coagulation screening with the functional assays, our study revealed increased fibrinolysis in SD plasma, while augmented fibrinolysis could be excluded, even at trough FXIII levels in congenital FXIII deficiency. In LPI derivatives of fibrin formation and fibrinolysis confirmed their abnormalities, also detected by in vivo markers. Enhanced TG and fibrinolysis were observed in SD plasma. SD treatment reduced the levels of single chain protein S, leading to a loss of both activated protein C -dependent and independent protein S activity, enhancing TG. In addition, the in vitro coagulation phenotype of SD plasma was abnormal due to increased fibrinolysis likely via diminished levels of a2-antiplasmin (a2-AP). Tranexamic acid abolished this fibrinolysis in vitro. Low levels of FXIII induced accelerated prothrombin conversion, leading to increased TG, possibly based on decreased fibrin antithrombin I (AT I) -like activity. According to AT I theory forming fibrin (= AT I) inhibits TG during coagulation process by sequestering thrombin. In our study, impaired fibrin formation induced both by lack of FXIII and GPRP administration in normal plasma enhanced TG, supporting the AT I hypothesis. In LPI, a metabolic disorder leading to renal insufficiency, we discovered impaired primary hemostasis and altered TG. Moreover, we detected decreased fibrin formation and markedly enhanced fibrinolysis that both related to altered plasminogen/a2-AP ratio and renal insufficiency. Platelet levels, function of primary hemostasis, fibrinogen and FXIII levels should be measured perioperatively and treated accordingly with platelet transfusions and fibrinogen and FXIII concentrates (in LPI). In summary, combining traditional and functional coagulation assays is beneficial. The altered coagulation phenotype of SD plasma should be acknowledged during transfusion therapy. Enhanced TG under low FXIII levels may guide FXIII replacement therapy. Our results concerning LPI further confirm the link between impaired coagulation and renal insufficiency.

KW - 3121 Internal medicine

M3 - Doctoral Thesis

SN - 978-951-51-4560-4

T3 - Dissertationes Scholae Doctoralis Ad Sanitatem Investigandam Universitatis Helsinkiensis

PB - Helsingin yliopisto

CY - Helsinki

ER -

Pitkänen H. Alterations and impact of thrombin generation and clot formation in solvent/detergent plasma, FXIII deficiency and lysinuric protein intolerance. Helsinki : Helsingin yliopisto, 2018. 81 p. (Dissertationes Scholae Doctoralis Ad Sanitatem Investigandam Universitatis Helsinkiensis; 68/2018 ).