Genetic testing of Mucopolysaccharidoses disease using multiplex PCR- based panels of STR markers: in silico analysis of novel mutations
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The Mucopolysaccharidoses (MPS) are group of inherited metabolic diseases caused by the deficiency of enzymes required to degrade glycosaminoglycans (GAGs) in the lysosomes. GAGs are sulfated polysaccharides involving repeating disaccharides, uronic acid and hexosamines including chondroitin sulfate (CS), dermatan sulfate (DS), heparan sulfate (HS) and keratan sulfate (KS). Hyaluronan is excluded in terms of being non-sulfated in the GAG family. Different types of mutations have been identified as the causative agent in all types of MPS. Herein, we planned to investigate the pathogenic mutations in different types of MPS including type I (IDUA gene), IIIA (SGSH) and IIIB (NAGLU) in the eight Iranian patients. Autozygosity mapping was performed to identify the potential pathogenic variants in these 8 patients indirectly with the clinical diagnosis of MPSs. so three panels of STR (Short Tandem Repeat) markres flanking IDUA, SGSH and NAGLU genes were selected for multiplex PCR amplification. Then in each family candidate gene was sequenced to identify the pathogenic mutation. Our study showed two novel mutations c.469 T > C and c.903C > G in the IDUA gene, four recurrent mutations: c.1A > C in IDUA, c.220C > T, c.1298G > A in SGSH gene and c.457G > A in the NAGLU gene. The c.1A > C in IDUA was the most common mutation in our study. In silico analysis were performed as well to predict the pathogenicity of the novel variants.
KeywordsMucopolysaccharidoses (MPS) Autozygosity mapping Linkage Mutation analysis Iran
Compliance with ethical standards
Conflict of interest
Mehdi Shafaat declares that there is no conflict of interest. Dr. Mehrdad Hashemi declares that there is no conflict of interest. Dr.Ahmad Majd declares that there is no conflict of interest. Dr. Maryam Abiri declares that there is no conflict of interest. Dr. Sirous zeinali declares that there is no conflict of interest.
Informed consent was received from 8 patients for participation in this study.
This study does not contain any animal study.
- Atceken, N., Ozgul, R. K., Yucel Yilmaz, D., Tokatli, A., Coskun, T., Sivri, H. S., . . . Karaca, M. (2016). Evaluation and identification of IDUA gene mutations in Turkishpatients with mucopolysaccharidosis type I. Turk J Med Sci, 46(2), 404–408. https://doi.org/10.3906/sag-1411-160 CrossRefGoogle Scholar
- Bertola, F., Filocamo, M., Casati, G., Mort, M., Rosano, C., Tylki-Szymanska, A., . . . Parini, R. (2011). IDUA mutational profiling of a cohort of 102 European patients with mucopolysaccharidosis type I: identification and characterization of 35 novel alpha-L-iduronidase (IDUA) alleles. Hum Mutat, 32(6), E2189–E2210. https://doi.org/10.1002/humu.21479 CrossRefGoogle Scholar
- Bunge, S., Kleijer, W. J., Steglich, C., Beck, M., Zuther, C., Morris, C. P., . . . Gal, A. (1994). Mucopolysaccharidosis type I: identification of 8 novel mutations and determination of the frequency of the two common alpha-L-iduronidase mutations (W402X and Q70X) among European patients. Hum Mol Genet, 3(6), 861–866CrossRefGoogle Scholar
- Chkioua, L., Khedhiri, S., Kassab, A., Bibi, A., Ferchichi, S., Froissart, R., . . . Miled, A. (2011). Molecular analysis of mucopolysaccharidosis type I in Tunisia: identification of novel mutation and eight novel polymorphisms. Diagn Pathol, 6, 39. https://doi.org/10.1186/1746-1596-6-39 CrossRefGoogle Scholar
- Duncan, F. J., Naughton, B. J., Zaraspe, K., Murrey, D. A., Meadows, A. S., Clark, K. R., . . . McCarty, D. M. (2015). Broad functional correction of molecular impairments by systemic delivery of scAAVrh74-hSGSH gene delivery in MPS IIIA mice. Mol Ther, 23(4), 638–647. https://doi.org/10.1038/mt.2015.9 CrossRefGoogle Scholar
- Hijikata, A., Raju, R., Keerthikumar, S., Ramabadran, S., Balakrishnan, L., Ramadoss, S. K., . . . Ohara, O. (2010). Mutation@a glance: an integrative web application for analysing mutations from human genetic diseases. DNA Res, 17(3), 197–208. https://doi.org/10.1093/dnares/dsq010 CrossRefGoogle Scholar
- Hinderer, C., Bell, P., Louboutin, J. P., Katz, N., Zhu, Y., Lin, G., . . . Wilson, J. M. (2016). Neonatal tolerance induction enables accurate evaluation of gene therapy for MPS I in a canine model. Mol Genet Metab, 119(1–2), 124–130. https://doi.org/10.1016/j.ymgme.2016.06.006 CrossRefGoogle Scholar
- Huh, H. J., Seo, J. Y., Cho, S. Y., Ki, C. S., Lee, S. Y., Kim, J. W., . . . Jin, D. K. (2013). The first Korean case of mucopolysaccharidosis IIIC (Sanfilippo syndrome type C) confirmed by biochemical and molecular investigation. Ann Lab Med, 33(1), 75–79. https://doi.org/10.3343/alm.2013.33.1.75 CrossRefGoogle Scholar
- Jansen, A. C., Cao, H., Kaplan, P., Silver, K., Leonard, G., De Meirleir, L., . . . Andermann, E. (2007). Sanfilippo syndrome type D: natural history and identification of 3 novel mutations in the GNS gene. Arch Neurol, 64(11), 1629–1634. https://doi.org/10.1001/archneur.64.11.1629 CrossRefGoogle Scholar
- Kim, C., Kwak, M. J., Cho, S. Y., Ko, A. R., Rheey, J., Kwon, J. Y., . . . Jin, D. K. (2015). Decreased performance in IDUA knockout mouse mimic limitations of joint function and locomotion in patients with hurler syndrome. Orphanet J Rare Dis, 10, 121. https://doi.org/10.1186/s13023-015-0337-3 CrossRefGoogle Scholar
- Lum, S. H., Miller, W. P., Jones, S., Poulton, K., Ogden, W., Lee, H., . . . Wynn, R. F. (2017a). Changes in the incidence, patterns and outcomes of graft failure following hematopoietic stem cell transplantation for hurler syndrome. Bone Marrow Transplant, 52(6), 846–853. https://doi.org/10.1038/bmt.2017.5 CrossRefGoogle Scholar
- Lum, S. H., Stepien, K. M., Ghosh, A., Broomfield, A., Church, H., Mercer, J., . . . Wynn, R. (2017b). Long term survival and cardiopulmonary outcome in children with hurler syndrome after haematopoietic stem cell transplantation. J Inherit Metab Dis, 40(3), 455–460. https://doi.org/10.1007/s10545-017-0034-6 CrossRefGoogle Scholar
- Muenzer J (2011) Overview of the mucopolysaccharidoses. Rheumatology (Oxford) 50(Suppl 5):v4–v12. https://doi.org/10.1093/rheumatology/ker394
- Oussoren E, Keulemans J, van Diggelen OP, Oemardien LF, Timmermans RG, van der Ploeg AT, Ruijter GJ (2013) Residual alpha-L-iduronidase activity in fibroblasts of mild to severe Mucopolysaccharidosis type I patients. Mol Genet Metab 109(4):377–381. https://doi.org/10.1016/j.ymgme.2013.05.016 CrossRefGoogle Scholar
- Prommajan K, Ausavarat S, Srichomthong C, Puangsricharern V, Suphapeetiporn K, Shotelersuk V (2011) A novel p.E276K IDUA mutation decreasing alpha-L-iduronidase activity causes mucopolysaccharidosis type I. Mol Vis 17:456–460Google Scholar
- Rodgers NJ, Kaizer AM, Miller WP, Rudser KD, Orchard PJ, Braunlin EA (2017) Mortality after hematopoietic stem cell transplantation for severe mucopolysaccharidosis type I: the 30-year University of Minnesota experience. J Inherit Metab Dis 40(2):271–280. https://doi.org/10.1007/s10545-016-0006-2 CrossRefGoogle Scholar
- Shafaat, M., Alaee, M. R., Rahmanifar, A., Setoodeh, A., Razzaghy-Azar, M., Bagherian, H., . . . Zeinali, S. (2018). Autozygosity mapping of methylmalonic acidemia associated genes by short tandem repeat markers facilitates the identification of five novel mutations in an Iranian patient cohort. Metab Brain Dis, 33(5), 1689–1697. https://doi.org/10.1007/s11011-018-0277-4 CrossRefGoogle Scholar
- Tebani, A., Abily-Donval, L., Schmitz-Afonso, I., Heron, B., Piraud, M., Ausseil, J., . . . Bekri, S. (2018). Unveiling metabolic remodeling in mucopolysaccharidosis type III through integrative metabolomics and pathway analysis. J Transl Med, 16(1), 248. https://doi.org/10.1186/s12967-018-1625-1 CrossRefGoogle Scholar
- Terlato NJ, Cox GF (2003) Can mucopolysaccharidosis type I disease severity be predicted based on a patient's genotype? A comprehensive review of the literature. Genet Med 5(4):286–294. https://doi.org/10.1097/01.gim.0000078027.83236.49 CrossRefGoogle Scholar
- Valenzano KJ, Khanna R, Powe AC, Boyd R, Lee G, Flanagan JJ, Benjamin ER (2011) Identification and characterization of pharmacological chaperones to correct enzyme deficiencies in lysosomal storage disorders. Assay Drug Dev Technol 9(3):213–235. https://doi.org/10.1089/adt.2011.0370 CrossRefGoogle Scholar
- Valstar, M. J., Bruggenwirth, H. T., Olmer, R., Wevers, R. A., Verheijen, F. W., Poorthuis, B. J., . . . Wijburg, F. A. (2010). Mucopolysaccharidosis type IIIB may predominantly present with an attenuated clinical phenotype. J Inherit Metab Dis, 33(6), 759–767. https://doi.org/10.1007/s10545-010-9199-y CrossRefGoogle Scholar
- Vazna, A., Beesley, C., Berna, L., Stolnaja, L., Myskova, H., Bouckova, M., . . . Dvorakova, L. (2009). Mucopolysaccharidosis type I in 21 Czech and Slovak patients: mutation analysis suggests a functional importance of C-terminus of the IDUA protein. Am J Med Genet A, 149A(5), 965–974. https://doi.org/10.1002/ajmg.a.32812 CrossRefGoogle Scholar