Activated factor VII (FVIIa) forms a complex with Tissue factor (TF), thus triggering blood coagulation through the so-called FVII-TF pathway, whose constitutive inhibitor is the Tissue Factor Pathway Inhibitor (TFPI). FVII is a vitamin K-dependent serine protease, homologous to other coagulation factors and TF is a transmembrane glycoprotein that, when exposed, acts as the cellular receptor and essential cofactor for FVIIa. Although polymorphisms within FVII, TF and TFPI genes have been described to affect plasma/cellular levels of factors, it is not known how such gene variations could interact one another and with environmental components, and influence disease risk. The project is aimed at elucidating the biological modulation by genetic and epigenetic components of functional levels of components of the TF-FVII pathway, FVII, Tissue Factor (the cofactor) and TFPI (the specific inhibitor). We plan the cooperation between two units with shared and documented records in the proposed investigation field, the study of the initiation of blood coagulation in order to understand the molecular bases of bleeding and of thrombosis, the latter being a major feature of acute coronary syndromes, including myocardial infarction (MI). We hypothesize that the modulation of the interaction among the serine protease, cofactor and inhibitor in the initiation complex of coagulation could modulate the risk of bleeding or thrombosis. The human model will be integrated with mouse models to dissect poorly investigated modulators: 1) promoter DNA methylation, the main epigenetic feature of DNA that regulates gene expression. We hypothesize that the modulation of expression associated with polymorphisms may be also related to promoter methylation. Environment-diet factors play also a significant part in determining TF-FVII pathway proteins levels, and folate status, in association with a polymorphic enzyme responsible for the methyl group supply, affects DNA methylation status in a gene-nutrient interaction fashion. Nutrients that can affect DNA methylation status may modulate the relationship between polymorphisms and promoter methylation in TF-FVII pathway genes and affect gene expression and proteins levels. 2) circadian rhythms, which could cause ample daily functional variations and could explain marked diurnal variations in frequencies of hemorrhagic and thromboembolic events in humans. For this study the research group includes a member with specific expertise in the field, and particularly in animal models. The analysis of functional levels associated with mutations and functional polymorphisms will be favoured by appropriately designed assays which will include the determination of microvesicle associated TF, a form of intra-vascular TF important in both the initiation and propagation phases of coagulation. We plan to use plasma and reconstituted systems for the study of recombinant protein function, and cellular models to investigate expression and regulatory mechanisms. International co-operation to the project is provided for 1) recruitment of subjects with FVII deficiency (the PI is a member of the steering committee of the International Registry of FVII deficiency), 2) methodological stages of post-doctoral fellows in European or USA laboratories. Moreover, the researcher with a major role in the project for the investigation of epigenetic features has recently joined the group moving from a laboratory with a leading role in the study of gene-nutrient interactions. The biological achievements of the proposed research project could provide elements to design individual-oriented preventive or therapeutic approaches.International co-operation to the project is provided for 1) recruitment of subjects with FVII deficiency (the PI is a member of the steering committee of the International Registry of FVII deficiency), 2) methodological stages of post-doctoral fellows in European or USA laboratories. Moreover, the researcher with a major role in the project for the investigation of epigenetic features has recently joined the group moving from a laboratory with a leading role in the study of gene-nutrient interactions. The biological achievements of the proposed research project could provide elements to design individual-oriented preventive or therapeutic approaches.
Progetto PRIN: Modulation of Factor VII-Tissue Factor pathway in haemorrhagic and thrombotic diseases
BERNARDI, Francesco
2004
Abstract
Activated factor VII (FVIIa) forms a complex with Tissue factor (TF), thus triggering blood coagulation through the so-called FVII-TF pathway, whose constitutive inhibitor is the Tissue Factor Pathway Inhibitor (TFPI). FVII is a vitamin K-dependent serine protease, homologous to other coagulation factors and TF is a transmembrane glycoprotein that, when exposed, acts as the cellular receptor and essential cofactor for FVIIa. Although polymorphisms within FVII, TF and TFPI genes have been described to affect plasma/cellular levels of factors, it is not known how such gene variations could interact one another and with environmental components, and influence disease risk. The project is aimed at elucidating the biological modulation by genetic and epigenetic components of functional levels of components of the TF-FVII pathway, FVII, Tissue Factor (the cofactor) and TFPI (the specific inhibitor). We plan the cooperation between two units with shared and documented records in the proposed investigation field, the study of the initiation of blood coagulation in order to understand the molecular bases of bleeding and of thrombosis, the latter being a major feature of acute coronary syndromes, including myocardial infarction (MI). We hypothesize that the modulation of the interaction among the serine protease, cofactor and inhibitor in the initiation complex of coagulation could modulate the risk of bleeding or thrombosis. The human model will be integrated with mouse models to dissect poorly investigated modulators: 1) promoter DNA methylation, the main epigenetic feature of DNA that regulates gene expression. We hypothesize that the modulation of expression associated with polymorphisms may be also related to promoter methylation. Environment-diet factors play also a significant part in determining TF-FVII pathway proteins levels, and folate status, in association with a polymorphic enzyme responsible for the methyl group supply, affects DNA methylation status in a gene-nutrient interaction fashion. Nutrients that can affect DNA methylation status may modulate the relationship between polymorphisms and promoter methylation in TF-FVII pathway genes and affect gene expression and proteins levels. 2) circadian rhythms, which could cause ample daily functional variations and could explain marked diurnal variations in frequencies of hemorrhagic and thromboembolic events in humans. For this study the research group includes a member with specific expertise in the field, and particularly in animal models. The analysis of functional levels associated with mutations and functional polymorphisms will be favoured by appropriately designed assays which will include the determination of microvesicle associated TF, a form of intra-vascular TF important in both the initiation and propagation phases of coagulation. We plan to use plasma and reconstituted systems for the study of recombinant protein function, and cellular models to investigate expression and regulatory mechanisms. International co-operation to the project is provided for 1) recruitment of subjects with FVII deficiency (the PI is a member of the steering committee of the International Registry of FVII deficiency), 2) methodological stages of post-doctoral fellows in European or USA laboratories. Moreover, the researcher with a major role in the project for the investigation of epigenetic features has recently joined the group moving from a laboratory with a leading role in the study of gene-nutrient interactions. The biological achievements of the proposed research project could provide elements to design individual-oriented preventive or therapeutic approaches.International co-operation to the project is provided for 1) recruitment of subjects with FVII deficiency (the PI is a member of the steering committee of the International Registry of FVII deficiency), 2) methodological stages of post-doctoral fellows in European or USA laboratories. Moreover, the researcher with a major role in the project for the investigation of epigenetic features has recently joined the group moving from a laboratory with a leading role in the study of gene-nutrient interactions. The biological achievements of the proposed research project could provide elements to design individual-oriented preventive or therapeutic approaches.I documenti in SFERA sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
