Contributions
Abstract: PB1973
Type: Publication Only
Background
- Pyruvate kinase (PK) deficiency is one of the most common causes of hereditary hemolytic anemia around the world with clinical manifestations varying from mild to severe hemolysis. Here we investigate PK-deficient cases referred to our laboratory for biological analysis of the unknown cause of hemolytic anemia.
Aims
To diagnose Pyruvate kinase deficiency (PKD) in hereditary non spherocytic hemolytic anemia cases in India and Genotype-phenotype correlations in PKD
Methods
Test for aaemoglobinopathies, RBC membrane protein defects and red cell enzyme activities were measured by standard methods; Molecular characterization of the PK gene mutations included restriction enzyme analysis, mutation scanning by DNA sequencing. In silico prediction using bioinformatics tools (SIFT, Polyphen2) was done and the molecular modeling were performed using PYMOL software.
Results
Total 21 families were identified in which 17 homozygous PKD cases identified and eighteen different PKLR mutations were found among which six are described for the first time (V294M, I282S, M373V, A423E, W525X and R559X) and 11 known mutations which includes (E315K, D331G, G332S, G358R, N393S,E407K, R486W, R479H, R498C, V506I , R510Q and IVS9 ds A-G +3 ) are associated with severe clinical presentation. All missense mutations were located in crucial domains of the molecule (catalytic site, cleft between the A and C domains, A/A' interface and have severely damaging effect leading to need of regular blood transfusion. The most frequent mutations in the Indian population appear to be V294M, followed by R479H, R510Q and D331G. Genotype-phenotype correlations for the novel mutations were investigated by three-dimensional structure analysis.
Conclusion
In India, the genetic heterogeneity of PKLR is still high but differs from that observed in the previous study carried out in 2009 and 2013. DNA sequencing was helping for genetic counseling and prenatal diagnosis.
Session topic: 29. Enzymopathies, membranopathies and other anemias
Keyword(s): Hemolytic anemia, mutation analysis
Abstract: PB1973
Type: Publication Only
Background
- Pyruvate kinase (PK) deficiency is one of the most common causes of hereditary hemolytic anemia around the world with clinical manifestations varying from mild to severe hemolysis. Here we investigate PK-deficient cases referred to our laboratory for biological analysis of the unknown cause of hemolytic anemia.
Aims
To diagnose Pyruvate kinase deficiency (PKD) in hereditary non spherocytic hemolytic anemia cases in India and Genotype-phenotype correlations in PKD
Methods
Test for aaemoglobinopathies, RBC membrane protein defects and red cell enzyme activities were measured by standard methods; Molecular characterization of the PK gene mutations included restriction enzyme analysis, mutation scanning by DNA sequencing. In silico prediction using bioinformatics tools (SIFT, Polyphen2) was done and the molecular modeling were performed using PYMOL software.
Results
Total 21 families were identified in which 17 homozygous PKD cases identified and eighteen different PKLR mutations were found among which six are described for the first time (V294M, I282S, M373V, A423E, W525X and R559X) and 11 known mutations which includes (E315K, D331G, G332S, G358R, N393S,E407K, R486W, R479H, R498C, V506I , R510Q and IVS9 ds A-G +3 ) are associated with severe clinical presentation. All missense mutations were located in crucial domains of the molecule (catalytic site, cleft between the A and C domains, A/A' interface and have severely damaging effect leading to need of regular blood transfusion. The most frequent mutations in the Indian population appear to be V294M, followed by R479H, R510Q and D331G. Genotype-phenotype correlations for the novel mutations were investigated by three-dimensional structure analysis.
Conclusion
In India, the genetic heterogeneity of PKLR is still high but differs from that observed in the previous study carried out in 2009 and 2013. DNA sequencing was helping for genetic counseling and prenatal diagnosis.
Session topic: 29. Enzymopathies, membranopathies and other anemias
Keyword(s): Hemolytic anemia, mutation analysis