Identification of six novel factor viii gene variants using next generation sequencing and molecular dynamics simulation

Faisal A Al-Allaf; Zainularifeen Abduljaleel; Neda M Bogari; Tarek M A Owaidah; Mohiuddin M Taher; Mohammad Athar; Ahmed Elsendiony; Halah Abalkhail; Ahmed Abdellatif; Wafa Elbjeirami; Abdellatif Bouazzaoui

doi:10.18388/abp.2018_2339

Faisal A Al-Allaf Faculty of Medicine, Umm Al-Qura University Science and Technology Unit, Umm Al-Qura University Molecular Diagnostics Unit, Department of Laboratory and Blood Bank, King Abdullah Medical City
Zainularifeen Abduljaleel Faculty of Medicine, Umm Al-Qura University Science and Technology Unit, Umm Al-Qura University
Neda M Bogari Faculty of Medicine, Umm Al-Qura University Science and Technology Unit, Umm Al-Qura University
Tarek M A Owaidah King Faisal Specialist Hospital and Research Center, Riyadh
Mohiuddin M Taher Faculty of Medicine, Umm Al-Qura University Science and Technology Unit, Umm Al-Qura University
Mohammad Athar Faculty of Medicine, Umm Al-Qura University Science and Technology Unit, Umm Al-Qura University
Ahmed Elsendiony Makkah Regional Laboratory, Makkah, Saudi Arabia
Halah Abalkhail King Faisal Specialist Hospital and Research Center, Riyadh
Ahmed Abdellatif Faculty of Pharmacy, Qassim University, Qassim, Saudi Arabia
Wafa Elbjeirami King Abdullah Medical City, Makkah
Abdellatif Bouazzaoui Faculty of Medicine, Umm Al-Qura University Science and Technology Unit, Umm Al-Qura University

DOI: https://doi.org/10.18388/abp.2018_2339

Abstract

Hemophilia A is an X-linked recessive hemorrhagic disorder caused by variants in the F8 gene. To identify known and novel causative variants in hemophilia A, we have carried out genetic analysis among Saudi patients. Twenty-one patients, who were negative for inv-1/inv-22, were selected for analysis by next generation sequencing, thereafter confirmed by Sanger sequencing. In addition, the functionality and structural changes in the variant proteins were assessed using Molecular dynamics (MD) simulation and compared with wild-type and native proteins. In the samples we analyzed, we found 10 variants in 12 individuals; among them, five were novel and five were previously reported. The novel variants were located at positions: c.6130_6131insC, c.5815G>C, c.5493C>G, c.3734_3740delinsATTTCT and c.3744A>T. With the exception of one variant which was silent, the MD simulation revealed that the observed variants were causing severe structural changes when compared to the native protein and resulted in a loss of the protein’s function. The MD analysis is in line with clinical data of patients who had <1% Factor VIII levels (severe hemophilia) with episodic bleeding, and were on more than one treatment. Moreover, some patients presented with chronic joint disability. These results will enrich the spectrum of variants and enlarge the factor VIII protein’s database in the Saudi Arabian population.

Author Biographies

Faisal A Al-Allaf, Faculty of Medicine, Umm Al-Qura University Science and Technology Unit, Umm Al-Qura University Molecular Diagnostics Unit, Department of Laboratory and Blood Bank, King Abdullah Medical City

Department of Medical Genetics

Assis Prof

Zainularifeen Abduljaleel, Faculty of Medicine, Umm Al-Qura University Science and Technology Unit, Umm Al-Qura University

Department of Medical Genetics

MS

Neda M Bogari, Faculty of Medicine, Umm Al-Qura University Science and Technology Unit, Umm Al-Qura University

Department of Medical Genetics

Assis Prof

Tarek M A Owaidah, King Faisal Specialist Hospital and Research Center, Riyadh

Department of Pathology and Laboratory Medicine

Prof.

Mohiuddin M Taher, Faculty of Medicine, Umm Al-Qura University Science and Technology Unit, Umm Al-Qura University

Department of Medical Genetics

Assis Prof

Mohammad Athar, Faculty of Medicine, Umm Al-Qura University Science and Technology Unit, Umm Al-Qura University

Department of Medical Genetics

Assis Prof

Halah Abalkhail, King Faisal Specialist Hospital and Research Center, Riyadh

Department of Pathology and Laboratory Medicine

Wafa Elbjeirami, King Abdullah Medical City, Makkah

Molecular Diagnostics Unit, Department of Laboratory and Blood Bank

Assis Prof

Abdellatif Bouazzaoui, Faculty of Medicine, Umm Al-Qura University Science and Technology Unit, Umm Al-Qura University

Department of Medical Genetics

Assis Prof

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