Significance of two point mutations present in each HEXB allele of patients with adult GM2 gangliosidosis (Sandhoff disease) Homozygosity for the Ile207 → Val substitution is not associated with a clinical or biochemical phenotype

摘要

The molecular defects in the HEXB gene encoding the common β-subunit of lysosomal β-hexosaminidase A (β-Hex A, αβ) and β-Hex B (ββ) were investigated in a Portuguese family affected with late onset Sandhoff disease (GM2-gangliosidosis variant 0). This family comprised two unaffected daughters and three affected sibs who developed at about age 17 cerebellar ataxia and mental deficiency. Their parents were consanguineous and clinically asymptomatic. There was no detectable β-Hex B activity and a profound reduction in the activity of β-Hex A in the leukocytes and transformed lymphoid cell lines from the affected sibs. The expected intermediate values were observed in the parents as well as in one daughter and her children. Western analysis revealed the presence of reduced, but detectable amounts of mature β-chain protein in cell lysates from the probands and intermediate levels in the parents. Nucleotide sequencing of amplified, reverse-transcribed β-chain mRNA demonstrated the presence of two single point mutations: an A619 to G transition in exon 5 (Ile207 → Val), and a G1514 to A transition in exon 13 (Arg505 → Gln). Both of these two mutations have been previously linked to the adult form of Sandhoff disease in compound heterozygote patients. All three affected sibs were found to be homoallelic for both mutations. Interestingly, while the mother was heterozygous for each mutation, the father was homozygote for the A619 → G substitution and heterozygote for the G1514 → A transition. Since the father is homozygote for the A619 → G mutation but expresses a biochemical phenotype consistent with a carrier of Sandhoff disease and is clinically asymptomatic, this substitution is likely a neutral mutation. We confirmed this hypothesis by finding this transition present in 4 of 30 alleles from normal individuals. We conclude that homozygosity for the G1514 → A mutation is exclusively responsible for the adult form of Sandhoff disease in this family, and that the A619 → G substitution is not a deleterious mutation but rather a common HEXB polymorphism.