A Generic Assay to Detect Aberrant ARSB Splicing and mRNA Degradation for the Molecular Diagnosis of MPS VI
Identification and characterization of disease-associated variants in monogenic disorders is an important aspect of diagnosis, genetic counseling, prediction of disease severity, and development of therapy. However, the effects of disease-associated variants on pre-mRNA splicing and mRNA degradation...
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Elsevier
2020-12-01
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Series: | Molecular Therapy: Methods & Clinical Development |
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Online Access: | http://www.sciencedirect.com/science/article/pii/S2329050120301893 |
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doaj-aa2096434c614e02b965124eabcdc167 |
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record_format |
Article |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Mike Broeders Kasper Smits Busra Goynuk Esmee Oussoren Hannerieke J.M.P. van den Hout Atze J. Bergsma Ans T. van der Ploeg W.W.M. Pim Pijnappel |
spellingShingle |
Mike Broeders Kasper Smits Busra Goynuk Esmee Oussoren Hannerieke J.M.P. van den Hout Atze J. Bergsma Ans T. van der Ploeg W.W.M. Pim Pijnappel A Generic Assay to Detect Aberrant ARSB Splicing and mRNA Degradation for the Molecular Diagnosis of MPS VI Molecular Therapy: Methods & Clinical Development lysosomal disease splicing antisense oligonucleotides pseudo exon diagnosis nonsense mediated decay |
author_facet |
Mike Broeders Kasper Smits Busra Goynuk Esmee Oussoren Hannerieke J.M.P. van den Hout Atze J. Bergsma Ans T. van der Ploeg W.W.M. Pim Pijnappel |
author_sort |
Mike Broeders |
title |
A Generic Assay to Detect Aberrant ARSB Splicing and mRNA Degradation for the Molecular Diagnosis of MPS VI |
title_short |
A Generic Assay to Detect Aberrant ARSB Splicing and mRNA Degradation for the Molecular Diagnosis of MPS VI |
title_full |
A Generic Assay to Detect Aberrant ARSB Splicing and mRNA Degradation for the Molecular Diagnosis of MPS VI |
title_fullStr |
A Generic Assay to Detect Aberrant ARSB Splicing and mRNA Degradation for the Molecular Diagnosis of MPS VI |
title_full_unstemmed |
A Generic Assay to Detect Aberrant ARSB Splicing and mRNA Degradation for the Molecular Diagnosis of MPS VI |
title_sort |
generic assay to detect aberrant arsb splicing and mrna degradation for the molecular diagnosis of mps vi |
publisher |
Elsevier |
series |
Molecular Therapy: Methods & Clinical Development |
issn |
2329-0501 |
publishDate |
2020-12-01 |
description |
Identification and characterization of disease-associated variants in monogenic disorders is an important aspect of diagnosis, genetic counseling, prediction of disease severity, and development of therapy. However, the effects of disease-associated variants on pre-mRNA splicing and mRNA degradation are difficult to predict and often missed. Here we present a generic assay for unbiased identification and quantification of arylsulfatase B (ARSB) mRNA for molecular diagnosis of patients with mucopolysaccharidosis VI (MPS VI). We found that healthy control individuals have inefficient ARSB splicing because of natural skipping of exon 5 and inclusion of two pseudoexons in introns 5 and 6. Analyses of 12 MPS VI patients with 10 different genotypes resulted in identification of a 151-bp intron inclusion caused by the c.1142+2T>C variant and detection of low ARSB expression from alleles with the c.629A>G variant. A special case showed skipping of exon 4 and low ARSB expression. Although no disease-associated DNA variant could be identified in this patient, the molecular diagnosis could be made based on RNA. These results highlight the relevance of RNA-based analyses to establish a molecular diagnosis of MPS VI. We speculate that inefficient natural splicing of ARSB may be a target for therapy based on promotion of canonical splicing. |
topic |
lysosomal disease splicing antisense oligonucleotides pseudo exon diagnosis nonsense mediated decay |
url |
http://www.sciencedirect.com/science/article/pii/S2329050120301893 |
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doaj-aa2096434c614e02b965124eabcdc1672020-12-11T04:21:53ZengElsevierMolecular Therapy: Methods & Clinical Development2329-05012020-12-0119174185A Generic Assay to Detect Aberrant ARSB Splicing and mRNA Degradation for the Molecular Diagnosis of MPS VIMike Broeders0Kasper Smits1Busra Goynuk2Esmee Oussoren3Hannerieke J.M.P. van den Hout4Atze J. Bergsma5Ans T. van der Ploeg6W.W.M. Pim Pijnappel7Department of Pediatrics, Erasmus MC University Medical Center, Rotterdam, the Netherlands; Department of Clinical Genetics, Erasmus MC University Medical Center, Rotterdam, the Netherlands; Center for Lysosomal and Metabolic Diseases, Erasmus MC University Medical Center, 3015 GE Rotterdam, the NetherlandsDepartment of Pediatrics, Erasmus MC University Medical Center, Rotterdam, the Netherlands; Department of Clinical Genetics, Erasmus MC University Medical Center, Rotterdam, the Netherlands; Center for Lysosomal and Metabolic Diseases, Erasmus MC University Medical Center, 3015 GE Rotterdam, the NetherlandsDepartment of Pediatrics, Erasmus MC University Medical Center, Rotterdam, the Netherlands; Department of Clinical Genetics, Erasmus MC University Medical Center, Rotterdam, the Netherlands; Center for Lysosomal and Metabolic Diseases, Erasmus MC University Medical Center, 3015 GE Rotterdam, the NetherlandsDepartment of Pediatrics, Erasmus MC University Medical Center, Rotterdam, the Netherlands; Center for Lysosomal and Metabolic Diseases, Erasmus MC University Medical Center, 3015 GE Rotterdam, the NetherlandsDepartment of Pediatrics, Erasmus MC University Medical Center, Rotterdam, the Netherlands; Center for Lysosomal and Metabolic Diseases, Erasmus MC University Medical Center, 3015 GE Rotterdam, the NetherlandsDepartment of Pediatrics, Erasmus MC University Medical Center, Rotterdam, the Netherlands; Department of Clinical Genetics, Erasmus MC University Medical Center, Rotterdam, the Netherlands; Center for Lysosomal and Metabolic Diseases, Erasmus MC University Medical Center, 3015 GE Rotterdam, the NetherlandsDepartment of Pediatrics, Erasmus MC University Medical Center, Rotterdam, the Netherlands; Center for Lysosomal and Metabolic Diseases, Erasmus MC University Medical Center, 3015 GE Rotterdam, the NetherlandsDepartment of Pediatrics, Erasmus MC University Medical Center, Rotterdam, the Netherlands; Department of Clinical Genetics, Erasmus MC University Medical Center, Rotterdam, the Netherlands; Center for Lysosomal and Metabolic Diseases, Erasmus MC University Medical Center, 3015 GE Rotterdam, the Netherlands; Corresponding author: W.W.M. Pim Pijnappel, Erasmus MC University Medical Center, Wytemaweg 80, 3015 CN Rotterdam, the Netherlands.Identification and characterization of disease-associated variants in monogenic disorders is an important aspect of diagnosis, genetic counseling, prediction of disease severity, and development of therapy. However, the effects of disease-associated variants on pre-mRNA splicing and mRNA degradation are difficult to predict and often missed. Here we present a generic assay for unbiased identification and quantification of arylsulfatase B (ARSB) mRNA for molecular diagnosis of patients with mucopolysaccharidosis VI (MPS VI). We found that healthy control individuals have inefficient ARSB splicing because of natural skipping of exon 5 and inclusion of two pseudoexons in introns 5 and 6. Analyses of 12 MPS VI patients with 10 different genotypes resulted in identification of a 151-bp intron inclusion caused by the c.1142+2T>C variant and detection of low ARSB expression from alleles with the c.629A>G variant. A special case showed skipping of exon 4 and low ARSB expression. Although no disease-associated DNA variant could be identified in this patient, the molecular diagnosis could be made based on RNA. These results highlight the relevance of RNA-based analyses to establish a molecular diagnosis of MPS VI. We speculate that inefficient natural splicing of ARSB may be a target for therapy based on promotion of canonical splicing.http://www.sciencedirect.com/science/article/pii/S2329050120301893lysosomal diseasesplicingantisense oligonucleotidespseudo exondiagnosisnonsense mediated decay |