Contributions
Abstract: EP1146
Type: E-Poster Presentation
Session title: Platelet disorders
Background
Immune thrombocytopenia (ITP) is a relatively common acquired bleeding disorder in childhood. ITP is characterized by a low circulating platelet count due to the production of autoantibodies against platelet glycoproteins, followed by their destruction via the reticuloendothelial system. Apoptosis is a physiologic mechanism of programmed cell death, crucial for controlling the cell number during development and for the entire lifespan of an organism. Deregulation of the apoptosis signalling may compromise the homeostasis of an organism and cause a wide range of diseases. Fas is a transmembrane molecule belonging to the tumor necrosis factor (TNF) receptor superfamily. Fas ligation by FasL induces programmed cell death in several lymphoid cell lines and may play a role in immune response control, lymphocyte life span regulation and induction of peripheral tolerance. Caspases are cysteine proteases that play a central role in apoptosis. Caspase-8 and caspase-10 are the first enzymes in the proteolytic cascade that are activated by Fas ligand and TNF. According to the literature, mutations in either CASP8 or CASP10 influence immune surveillance of malignancies and cause lymphoproliferation (ALPS).
Aims
The aim of this study was to identify whether polymorphisms of FAS, CASP8 and CASP10 play a role in the pathophysiology of ITP in children and adolescents.
Methods
In this study 30 children and adolescents with ITP and 60 healthy individuals as control group were included. At first, extraction of the DNA from peripheral blood cells was performed and then analysed by PCR. The PCR was followed by digestion using selected restricted enzymes in order to genotype individuals for the SNPs under study. For the FAS (rs2234767) and CASP8 (rs1045485), the restriction enzyme BstUI was applied while for the CASP10 (rs13006529), the enzyme SspI. Digestion products were analysed by agarose gel electrophoresis and statistical analysis was performed using the GraphPad Prism (GraphPad Software, San Diego, CA, USA) program.
Results
Statistical analysis of data showed that allele T of CASP10 rs13006529 SNP was associated with the diagnosis of ITP. However, no association for the studied SNPs of FAS and CASP8 genes was detected.
Conclusion
Allele T of CASP10 rs13006529 SNP appeared has a statistically significant difference between patients and controls, thus suggesting its role in the development of ITP. Its effect may be related to the role of caspase 10 in the endogenous pathway of apoptosis as well as its involvement in the process of autophagy. As regards with the FAS (rs2234767) and CASP8 (rs1045485) SNPs no association with ITP was observed. However, these results are considered as early due to the small number of samples examined. Further analysis of a higher number of samples is needed in order to clarify the role of the SNPs under study in children and adolescents with ITP.
Keyword(s): Caspase, Childhood, Gene polymorphism, Thrombocytopenia
Abstract: EP1146
Type: E-Poster Presentation
Session title: Platelet disorders
Background
Immune thrombocytopenia (ITP) is a relatively common acquired bleeding disorder in childhood. ITP is characterized by a low circulating platelet count due to the production of autoantibodies against platelet glycoproteins, followed by their destruction via the reticuloendothelial system. Apoptosis is a physiologic mechanism of programmed cell death, crucial for controlling the cell number during development and for the entire lifespan of an organism. Deregulation of the apoptosis signalling may compromise the homeostasis of an organism and cause a wide range of diseases. Fas is a transmembrane molecule belonging to the tumor necrosis factor (TNF) receptor superfamily. Fas ligation by FasL induces programmed cell death in several lymphoid cell lines and may play a role in immune response control, lymphocyte life span regulation and induction of peripheral tolerance. Caspases are cysteine proteases that play a central role in apoptosis. Caspase-8 and caspase-10 are the first enzymes in the proteolytic cascade that are activated by Fas ligand and TNF. According to the literature, mutations in either CASP8 or CASP10 influence immune surveillance of malignancies and cause lymphoproliferation (ALPS).
Aims
The aim of this study was to identify whether polymorphisms of FAS, CASP8 and CASP10 play a role in the pathophysiology of ITP in children and adolescents.
Methods
In this study 30 children and adolescents with ITP and 60 healthy individuals as control group were included. At first, extraction of the DNA from peripheral blood cells was performed and then analysed by PCR. The PCR was followed by digestion using selected restricted enzymes in order to genotype individuals for the SNPs under study. For the FAS (rs2234767) and CASP8 (rs1045485), the restriction enzyme BstUI was applied while for the CASP10 (rs13006529), the enzyme SspI. Digestion products were analysed by agarose gel electrophoresis and statistical analysis was performed using the GraphPad Prism (GraphPad Software, San Diego, CA, USA) program.
Results
Statistical analysis of data showed that allele T of CASP10 rs13006529 SNP was associated with the diagnosis of ITP. However, no association for the studied SNPs of FAS and CASP8 genes was detected.
Conclusion
Allele T of CASP10 rs13006529 SNP appeared has a statistically significant difference between patients and controls, thus suggesting its role in the development of ITP. Its effect may be related to the role of caspase 10 in the endogenous pathway of apoptosis as well as its involvement in the process of autophagy. As regards with the FAS (rs2234767) and CASP8 (rs1045485) SNPs no association with ITP was observed. However, these results are considered as early due to the small number of samples examined. Further analysis of a higher number of samples is needed in order to clarify the role of the SNPs under study in children and adolescents with ITP.
Keyword(s): Caspase, Childhood, Gene polymorphism, Thrombocytopenia