Novel R225C variant identified in the <i>HGD</i> gene in Jordanian patients with alkaptonuria

Nesrin R. Mwafi; Dema A. Ali; Raida W. Khalil; Ibrahim N. Alsbou'; Ahmad M. Saraireh; Nesrin R. Mwafi; Dema A. Ali; Raida W. Khalil; Ibrahim N. Alsbou'; Ahmad M. Saraireh

doi:10.3934/molsci.2021005

AIMS Molecular Science

2021, Volume 8, Issue 1: 60-75. doi: 10.3934/molsci.2021005

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Research article

Novel R225C variant identified in the HGD gene in Jordanian patients with alkaptonuria

1.
Department of biochemistry and molecular biology, Faculty of Medicine, Mutah University, Al-karak 61710, Jordan
2.
Cell Therapy Center, The University of Jordan, Amman 11942, Jordan
3.
Department of Biotechnology and Genetics Engineering, University of Philadelphia, Amman 19392, Jordan
4.
Medical Laboratory Science, Faculty of Science, Mutah University, Al-karak 61710, Jordan
5.
Faculty of Medicine, Mutah University, Al-karak 61710, Jordan

Received: 18 December 2020 Accepted: 18 February 2021 Published: 25 February 2021

Alkaptonuria (AKU) is a rare metabolic disease which is inherited as an autosomal recessive trait. It is characterized by the accumulation of homogentisic acid over time in various tissues of the body particularly connective tissues. This genetic disease is caused by mutation of the Homogentisate 1,2-dioxygenase (HGD) gene which encodes for enzyme essential for the catabolism of phenylalanine and tyrosine. The aim of the present study is to investigate variant types in Jordanian patients with alkaptonuria. Genomic DNA was extracted from whole blood samples of the participated AKU family members (n = 23). The 14 exons of HGD gene for the proband were amplified using specific PCR primers. The sequenced data were analysed and the pathogenicity of the identified variants were predicted using the online bioinformatics programs: PolyPhen2, SIFT and Mutation taster. The analysis showed that the proband was compound heterozygous for the missense mutations A122V and R225C found within E6 and E10, respectively. R225C variant is novel and the genotyping of the family members indicated that HGD^A122V and HGD^R225C alleles were fully segregated. Moreover, the cousins of the proband who are AKU patients inherited the homozygous pattern of the novel mutation. This study extends the pathogenic mutations spectrum of the HGD gene. It identified the novel mutation R225C and at the same time confirmed the high prevalence of the founder mutation A122V in Jordanian AKU patients.
- alkaptonuria,
- autosomal recessive,
- compound heterozygous,
- Homogentisate 1,2-dioxygenase gene,
- missense mutation
Citation: Nesrin R. Mwafi, Dema A. Ali, Raida W. Khalil, Ibrahim N. Alsbou', Ahmad M. Saraireh. Novel R225C variant identified in the HGD gene in Jordanian patients with alkaptonuria[J]. AIMS Molecular Science, 2021, 8(1): 60-75. doi: 10.3934/molsci.2021005

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Abstract

Alkaptonuria (AKU) is a rare metabolic disease which is inherited as an autosomal recessive trait. It is characterized by the accumulation of homogentisic acid over time in various tissues of the body particularly connective tissues. This genetic disease is caused by mutation of the Homogentisate 1,2-dioxygenase (HGD) gene which encodes for enzyme essential for the catabolism of phenylalanine and tyrosine. The aim of the present study is to investigate variant types in Jordanian patients with alkaptonuria. Genomic DNA was extracted from whole blood samples of the participated AKU family members (n = 23). The 14 exons of HGD gene for the proband were amplified using specific PCR primers. The sequenced data were analysed and the pathogenicity of the identified variants were predicted using the online bioinformatics programs: PolyPhen2, SIFT and Mutation taster. The analysis showed that the proband was compound heterozygous for the missense mutations A122V and R225C found within E6 and E10, respectively. R225C variant is novel and the genotyping of the family members indicated that HGD^A122V and HGD^R225C alleles were fully segregated. Moreover, the cousins of the proband who are AKU patients inherited the homozygous pattern of the novel mutation. This study extends the pathogenic mutations spectrum of the HGD gene. It identified the novel mutation R225C and at the same time confirmed the high prevalence of the founder mutation A122V in Jordanian AKU patients.

Abbreviations

AKU

Alkaptonuria

GC-MS

Gas Chromatography-Mass Spectrometry analysis

HGD

Homogentisate 1,2-dioxygenase

MLPA

multiplex ligation-dependent probe amplification

Acknowledgments

This study was funded by the Deanship of Scientific research and graduate studies at Philadelphia University in Jordan.

Conflict of interest

All authors declare no conflicts of interest in this paper.

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