- Letter to the Editor
- Open Access
Polymorphic variants (p.Ser141Ser and p.Arg737Gly) at the NAGLU gene are really indicative of pseudodeficiency alleles?
Italian Journal of Pediatrics volume 45, Article number: 60 (2019)
Filocamo et al. recently published a paper describing the presence of a pseudodeficiency allele, constituted by p.Ser141Ser and p.Arg737Gly polymorphisms at the NAGLU gene, which leads to a reduced level of the alpha-N-acetyl-D-glucosaminidase activity. Based on analysis performed in Brazilian patients, using a customized gene panel containing SGSH, NAGLU, HGSNAT and GNS we observed that p.Ser141Ser (rs659497) and p.Arg737Gly (rs86312) variants were present in homozygosis in all of our MPS IIIB patients and in the majority of MPS IIIA, IIIC and IIID patients, and there was no significant decrease of the alpha-N-acetyl-D-glucosaminidase enzyme activity in this group when compared with those without the “pseudodeficiency allele”. Thus, we suggest that these two variants are not producing a pseudodeficiency allele.
To the Editor
We read with great interest the article by Filocamo et al.  in the November 2018 issue of Italian Journal of Pediatrics. In this manuscript, the authors mention that a complication for molecular analysis of NAGLU gene (associated with Sanfilippo B syndrome or also called mucopolysaccharidoses type IIIB) is the presence of a pseudodeficiency allele, constituted by p.Ser141Ser and p.Arg737Gly polymorphisms simultaneously, which leads to a reduced level of the alpha-N-acetyl-D-iglucosamnidase activity.
We have been working on the diagnosis of mucopolysaccharidoses (MPS) over the past 35 years, and during this time, MPS diagnosis was confirmed in 1184 Brazilian patients, of which, 192 were affected by one of the four subtypes of MPS III . As part of our MPS III investigation, after detection of heparan sulphate glycosaminoglycan in urine, we performed different enzyme activity assays in leukocytes in order to reach the definitive diagnosis and to determine the MPS III subtype . Following this step, approximately 40% of our MPS III patients, were analyzed by targeted-next generation sequencing (NGS), using a customized gene panel containing SGSH, NAGLU, HGSNAT and GNS which was designed to simultaneous sequence the entire coding regions plus 20pb of intron-exon junction of these genes. Pathogenic variants were detected in all the cases. Therefore, we have extensive biochemical and molecular data of our group of patients.
Based in our data, p.Ser141Ser (rs659497) and p.Arg737Gly (rs86312) variants were present in homozygosis in all our MPS IIIB patients analyzed by NGS. For these patients, the enzyme activities were below the reference range, as they are clinically affected by MPS IIIB as proven by mutation analysis.
On the other hand, we have patients with other types of MPS III (IIIA, IIIC and IIID) where the same alterations that are said to cause the pseudodeficiency allele were present in homozygosis. These variants were detected because patients had clinical phenotype of MPS III and were analyzed in the same NGS panel used for the 4 types of MPS III. For those, the NAGLU enzyme activity was within the normal range when compared to normal controls as shown in Table 1.
These two variants were also present in homozygosis in one MPSIIIA patient from Turkey and two MPS IIIB from Ecuador.
For our group of MPS III patients, allele frequencies of p.Ser141Ser and p.Ar737Gly were 0.9706 and 0.9412, respectively. In addition, Exome Aggregation Consortium (ExAC) reported that frequency for p.Ser141Ser is 0.9947 and 0.9055 for p.Arg737Gly . Furthermore a Brazilian mutation database (ABraOM), which comprised the exomic variants of a cohort of 609 healthy Brazilian elderly, reported 602 homozygotes for p.Ser141Ser (0.9942) and 459 homozygotes for p.Arg737Gly (0.8686) .
Pseudodeficiency alleles are not rare in Lysosomal Disorders, including several MPS disorders [5,6,7,8,9,10], nevertheless the high frequency of both polymorphisms in worldwide populations and the fact that the presence of these alleles does not alter the MPS IIIB enzyme activity when present, suggests that these two variants are not producing a pseudodeficiency allele. Thus, no false positive results and no alteration on the protein product or changes in gene expression are detected in the presence of these two variants.
Exome Aggregation Consortium
- GNS :
- HGSNAT :
- NAGLU :
Next Generation Sequencing
- SGSH :
Filocamo M, Tomanin R, Bertola F, Morrone A. Biochemical and molecular analysis in mucopolysaccharidoses: what a paediatrician must know. Ital J Pediatr. 2018;44(S2):129 Available from: http://www.ncbi.nlm.nih.gov/pubmed/30442161. [cited 2019 Mar 7].
Giugliani R, Federhen A, Michelin-Tirelli K, Riegel M, Burin M. Relative frequency and estimated minimal frequency of lysosomal storage diseases in Brazil: report from a reference laboratory. Genet Mol Biol. 2017;40(1):31–9 Available from: http://www.ncbi.nlm.nih.gov/pubmed/28304074. [cited 2018 May 22].
Lek M, Karczewski KJ, Minikel EV, Samocha KE, Banks E, Fennell T, et al. Analysis of protein-coding genetic variation in 60,706 humans. Nature. 2016;536(7616):285–91 Available from: http://www.nature.com/articles/nature19057. [cited 2018 Dec 12].
Naslavsky MS, Yamamoto GL, de Almeida TF, Ezquina SAM, Sunaga DY, Pho N, et al. Exomic variants of an elderly cohort of Brazilians in the ABraOM database. Hum Mutat. 2017;38(7):751–63 Available from: http://doi.wiley.com/10.1002/humu.23220. [cited 2018 Jun 22].
Ricketts MH, Goldman D, Long JC, Manowitz P. Arylsulfatase a pseudodeficiency-associated mutations: population studies and identification of a novel haplotype. Am J Med Genet. 1996;67(4):387–92 Available from: http://www.ncbi.nlm.nih.gov/pubmed/8837707. [cited 2018 Jun 11].
Aronovich EL, Pan D, Whitley CB. Molecular genetic defect underlying alpha-L-iduronidase pseudodeficiency. Am J Hum Genet. 1996;58(1):75–85 Available from: http://www.ncbi.nlm.nih.gov/pubmed/8554071. [cited 2018 Jun 11].
Gort L, Santamaria R, Grinberg D, Vilageliu L, Chabás A. Identification of a novel pseudodeficiency allele in the GLB1 gene in a carrier of GM1 gangliosidosis. Clin Genet. 2007;72(2):109–11 Available from: http://doi.wiley.com/10.1111/j.1399-0004.2007.00843.x. [cited 2018 Jun 11].
Chabas A, Giros ML, Guardiola A. Low beta-glucuronidase activity in a healthy member of a family with mucopolysaccharidosis VII. J Inherit Metab Dis. 1991;14(6):908–14 Available from: http://www.ncbi.nlm.nih.gov/pubmed/1779649. [cited 2018 Jun 11].
Sandberg S, Deanching M, Hoganson G, Wenger D, Whitley C. Pseudo-deficiency allele of the N-acetylgalactosamine-4-sulfatase gene identified in a family with Maroteaux–Lamy syndrome (mucopolysaccharidosis type VI). Mol Genet Metab. 2008;93(2):34 Available from: http://linkinghub.elsevier.com/retrieve/pii/S1096719207005343. [cited 2018 Jun 26].
Vervoort R, Islam MR, Sly W, Chabas A, Wevers R, de Jong J, et al. A pseudodeficiency allele (D152N) of the human beta-glucuronidase gene. Am J Hum Genet. 1995;57(4):798–804 Available from: http://www.ncbi.nlm.nih.gov/pubmed/7573038. [cited 2018 Jun 26].
This study was partially supported by FIPE-HCPA (grant # 2013–0254) and Brazilian Institute for Population Medical Genetics (INAGEMP).
DRM is supported by CAPES post-doc scholarship. SLS receives a Research Grant from CNPq.
Availability of data and materials
Data sharing is not applicable to this article as no datasets were generated or analysed during the current study.
Ethics approval and consent to participate
This study was approved by the institutional ethics committee of Hospital de Clínicas de Porto Alegre, Brazil which is recognized by the Office for Human Research Protections as an Institutional Review Board (IRB0000921). All patients provided written informed consent (protocol number 2003–0066).
Consent for publication
The authors declare that they have no competing interests.
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