Future clinical and biochemical predictions of Fabry disease in females by methylation studies of the GLA gene

Mohammad Arif Hossain; Chen Wu; Hiroko Yanagisawa; Takashi Miyajima; Keiko Akiyama; Yoshikatsu Eto

doi:10.1016/j.ymgmr.2019.100497

Future clinical and biochemical predictions of Fabry disease in females by methylation studies of the GLA gene

Mol Genet Metab Rep. 2019 Jul 24:20:100497. doi: 10.1016/j.ymgmr.2019.100497. eCollection 2019 Sep.

Authors

Mohammad Arif Hossain^{1

2}, Chen Wu¹, Hiroko Yanagisawa¹, Takashi Miyajima¹, Keiko Akiyama¹, Yoshikatsu Eto^{1

2}

Affiliations

¹ Advanced Clinical Research Center, Institute of Neurological Disorders, Kawasaki, Kanagawa, Japan.
² Department of Gene Therapy, Institute for DNA Medicine, The Jikei University School of Medicine, Tokyo, Japan.

Abstract

Fabry disease is an X-linked lysosomal storage disorder caused by a deficiency of α-galactosidase A (α-gal A). The clinical variability of the phenotypes of Fabry disease in females is still poorly understood. The degree of aberrant methylation of non-mutated alleles is thought to have significant effects on X-chromosome inactivation (XCI). We previously reported that one heterozygous Fabry female showing classical phenotypes had complete methylation of the non-mutated allele of the GLA gene. In this report, we summarized 36 heterozygous females with a clinical severity score based on the FAbry STabilization indEX (FASTEX). We measured their α-gal A activity and plasma/ serum globotriaosylsphingosine (lyso-Gb3) accumulation and performed electron microscopy of skin biopsies. We analyzed the methylation-sensitive restriction enzyme sites throughout the GLA gene, including the 5'UTR, and found a single SacII site and multiple HhaI and HpaII sites aggregated in exon 1 and the 5'UTR. One HpaII sequence in exon 7 was also detected as a methylation-sensitive site. With methylation-sensitive restriction enzymes, methylated and non-methylated alleles could be separated, and the ratio of the methylation was quantified. We found a clear correlation between the severity of the phenotype and lyso-Gb3 accumulation for heterozygous Fabry disease in females. Methylation of the non-mutated allele was also proportionately correlated to the clinical severity score measured by FASTEX.

Keywords: Fabry stabilization indEX; Gb3, globotriaosylceramide; Heterozygous Fabry females; Methylation; X-chromosome inactivation; XCI, X-chromosomal inactivation; lyso-Gb3, globotriaosylsphingosine; α-gal A, α-galactosidase A.