Factor VIII Intron 22 Inversion in Severe Hemophilia A Patients in Palestine.

Nael Sudqi Abu Hasan,Caesar Mahmoud Abu Arra,Fekri Samarah

doi:10.1155/2020/3428648

Nael Sudqi Abu Hasan, Caesar Mahmoud Abu Arra + Show 1 more

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https://doi.org/10.1155/2020/3428648

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Abstract

Background Hemophilia A is an X-linked recessive bleeding disorder caused by mutations in FVIII gene with an incidence of 1 in 5,000 to 10,000 live born males. The Inv22 mutation is a major cause of the disease worldwide, accounting for up to 40%–50% of severe FVIII mutations. The aim of the present study was to screen Inv22 of the FVIII gene in Palestinian patients with severe HA and reveal its role as a predisposing factor for the development of inhibitors. Materials and Methods A cohort of 77 HA individuals including 5 carrier females from 52 unrelated families registered at governmental hemophilia centers in the West Bank area of Palestine was investigated. The demographic data and the clinical history were retrieved from medical files. Molecular analysis of Inv22 mutation in severe HA (30 cases) from Palestine was performed using the subcycling polymerase reaction (S-PCR). FVIII coagulant activities were carried out on an aPTT-based 1-stage clotting assay. FVIII inhibitors were quantified using the Nijmegen modification of the Bethesda assay. Result Overall, 41.7% (30/72) of the studied cases were classified as having severe HA, 22.2% (16/72) had moderate HA, and 36.1% (26/72) had mild HA. Five randomly selected carrier mothers were screened for the Inv22 mutation to confirm its transmission to their sons. The Inv22 mutation was detected in 11 severe HA patients (36.6%). Among the severe HA patients with positive Inv22, 45.5% (5/11) had developed inhibitors. The current study showed that there was no association (p=0.53) between inhibitor development and the Inv22 mutation. Conclusion Findings on Inv22 are in agreement with worldwide reports, being a major genetic mutation in severe HA. The S-PCR is a simple, rapid, and cost-effective method for the diagnosis of Inv22 in severe HA patients. Although the Inv22 mutation was associated with 36.6% of severe HA phenotype cases, it was not a major predisposing factor for inhibitor formation.

Highlights

Hemophilia A (HA) is an X-linked recessive bleeding disorder and the second most common coagulation disorder with an incidence of 1 in 5,000 to 10,000 live born males [1]
Intron 22, which is caused by recombination between a sequence within intron 22 of the factor VIII (FVIII) gene and one of the two homologous regions telomeric to the gene, and intron 1 inversions are the most frequent molecular alterations found in severe HA patients (FVIII:C
All patients were tested for factor VIII activities that were carried out on a 1stage activated partial thromboplastin time (aPTT)-based clotting assay via the Stago blood coagulation analyzer (STA Compact Max; Diagnostica Stago, Asnieres-sur-Seine, France) according to the manufacturer’s instructions. 10 ml of peripheral blood samples was collected in two tubes: tripotassium ethylene-diaminetetraacetate (EDTA K3) and 3.2% trisodium citrate tubes (Greiner Bio-One, Austria)

Summary

Introduction

Hemophilia A (HA) is an X-linked recessive bleeding disorder and the second most common coagulation disorder with an incidence of 1 in 5,000 to 10,000 live born males [1]. Hemophilia A is an X-linked recessive bleeding disorder caused by mutations in FVIII gene with an incidence of 1 in 5,000 to 10,000 live born males. E Inv mutation is a major cause of the disease worldwide, accounting for up to 40%–50% of severe FVIII mutations. E aim of the present study was to screen Inv of the FVIII gene in Palestinian patients with severe HA and reveal its role as a predisposing factor for the development of inhibitors. Molecular analysis of Inv mutation in severe HA (30 cases) from Palestine was performed using the subcycling polymerase reaction (SPCR). E current study showed that there was no association (p 0.53) between inhibitor development and the Inv mutation. The Inv mutation was associated with 36.6% of severe HA phenotype cases, it was not a major predisposing factor for inhibitor formation

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