TY - JOUR
T1 - Child-onset thrombotic thrombocytopenic purpura caused by p.R498C and p.G259PfsX133 mutations in ADAMTS13
AU - Schelpe, An-Sofie
AU - Orlando, Christelle
AU - Ercig, Bogac
AU - Geeroms, Chloe
AU - Pareyn, Inge
AU - Vandeputte, Nele
AU - Pereira, Leydi Carolina
AU - Roose, Elien
AU - Fostier, Karel
AU - Nicolaes, Gerry A. F.
AU - Deckmyn, Hans
AU - De Meyer, Simon F.
AU - Vanhoorelbeke, Karen
AU - Jochmans, Kristin
PY - 2018/8/1
Y1 - 2018/8/1
N2 - IntroductionPatients suffering from congenital thrombotic thrombocytopenic purpura (cTTP) have a deficiency in ADAMTS13 due to mutations in their ADAMTS13 gene. ObjectiveThe aim of this study was to determine ADAMTS13 parameters (activity, antigen, and mutations), to investigate if the propositus suffered from child-onset cTTP, and to study the in vitro effect of the ADAMTS13 mutations. MethodsADAMTS13 activity and antigen were determined using the FRETS VWF73 assay and ELISA and ADAMTS13 mutations via sequencing of the exons. Mutant proteins were expressed in Chinese hamster ovary cells, and their expression was studied using fluorescence microscopy and ELISA. Molecular modeling was used to evaluate the effect of the mutations on ADAMTS13 structure and stability. ResultsThe propositus was diagnosed with cTTP at the age of 20. ADAMTS13 activity was below 10%, and 2 compound heterozygous mutations, the p.R498C point and the p.G259PfsX133 frameshift mutation, were identified. Expression of ADAMTS13 mutants revealed that the p.R498C and the p.G259PfsX133 mutation cause secretion and translation defects in vitro, respectively. Molecular modeling showed that the R498 intra-domain interactions are lacking in the p.R498C mutant, resulting in protein instability. ConclusionThe ADAMTS13 mutations result in a severe ADAMTS13 deficiency explaining the patient's phenotype.
AB - IntroductionPatients suffering from congenital thrombotic thrombocytopenic purpura (cTTP) have a deficiency in ADAMTS13 due to mutations in their ADAMTS13 gene. ObjectiveThe aim of this study was to determine ADAMTS13 parameters (activity, antigen, and mutations), to investigate if the propositus suffered from child-onset cTTP, and to study the in vitro effect of the ADAMTS13 mutations. MethodsADAMTS13 activity and antigen were determined using the FRETS VWF73 assay and ELISA and ADAMTS13 mutations via sequencing of the exons. Mutant proteins were expressed in Chinese hamster ovary cells, and their expression was studied using fluorescence microscopy and ELISA. Molecular modeling was used to evaluate the effect of the mutations on ADAMTS13 structure and stability. ResultsThe propositus was diagnosed with cTTP at the age of 20. ADAMTS13 activity was below 10%, and 2 compound heterozygous mutations, the p.R498C point and the p.G259PfsX133 frameshift mutation, were identified. Expression of ADAMTS13 mutants revealed that the p.R498C and the p.G259PfsX133 mutation cause secretion and translation defects in vitro, respectively. Molecular modeling showed that the R498 intra-domain interactions are lacking in the p.R498C mutant, resulting in protein instability. ConclusionThe ADAMTS13 mutations result in a severe ADAMTS13 deficiency explaining the patient's phenotype.
KW - ADAMTS13
KW - mutation
KW - thrombotic thrombocytopenic purpura
KW - VON-WILLEBRAND-FACTOR
KW - FACTOR-CLEAVING PROTEASE
KW - UPSHAW-SCHULMAN-SYNDROME
KW - MISSENSE MUTATION
KW - GENE
KW - DEFICIENCY
KW - PLASMA
KW - MICROANGIOPATHIES
KW - POLYMORPHISMS
KW - PREVALENCE
U2 - 10.1111/ejh.13094
DO - 10.1111/ejh.13094
M3 - Article
C2 - 29763513
SN - 0902-4441
VL - 101
SP - 191
EP - 199
JO - European Journal of Haematology
JF - European Journal of Haematology
IS - 2
ER -