Molecular characterization of a novel splicing mutation underlying mucopolysaccharidosis (MPS) type VI-Indirect proof of principle on its pathogenicity
Autor(a) principal: | |
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Data de Publicação: | 2020 |
Outros Autores: | , , , , , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
Texto Completo: | http://hdl.handle.net/10400.18/6457 |
Resumo: | Here, we present the molecular diagnosis of a patient with a general clinical suspicion of Mucopolysaccharidosis, highlighting the different tools used to perform its molecular characterization. In order to decrease the turnaround time for the final report and contribute to reduce the "diagnostic odyssey", which frequently afflicts affected families, the proband's sample was simultaneously screened for mutations in a number of lysosomal function-related genes with targeted next-generation sequencing (NGS) protocol. After variant calling, the most probable cause for disease was a novel ARSB intronic variant, c.1213+5G>T [IVS6+5G>T], detected in homozygosity. In general, homozygous or compound heterozygous mutations in the ARSB gene, underlie MPS type VI or Maroteaux-Lamy syndrome. Still, even though the novel c.1213+5G>T variant was easy to detect by both NGS and Sanger sequencing, only through indirect studies and functional analyses could we present proof of principle on its pathogenicity. Globally, this case reminds us that whenever a novel variant is detected, its pathogenicity must be carefully assessed before a definitive diagnosis is established, while highlighting alternative approaches that may be used to assess its effect in the absence RNA/cDNA sample(s) from the proband. This is particularly relevant for intronic variants such as the one here reported. Special attention will be given to the use of reporter minigene systems, which may be constructed/designed to dissect the effect of this sort of alterations, providing an insight into their consequences over the normal pre-mRNA splicing process of the affected gene. |
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Molecular characterization of a novel splicing mutation underlying mucopolysaccharidosis (MPS) type VI-Indirect proof of principle on its pathogenicityLysosomal Storage DisordersFunctional StudiesMucopolysaccharidosis Type VINext-generation SequencingMPS VIGene ARSBDoenças GenéticasDoenças Lisossomais de SobrecargaMutações de SplicingMucopolissacaridosesHere, we present the molecular diagnosis of a patient with a general clinical suspicion of Mucopolysaccharidosis, highlighting the different tools used to perform its molecular characterization. In order to decrease the turnaround time for the final report and contribute to reduce the "diagnostic odyssey", which frequently afflicts affected families, the proband's sample was simultaneously screened for mutations in a number of lysosomal function-related genes with targeted next-generation sequencing (NGS) protocol. After variant calling, the most probable cause for disease was a novel ARSB intronic variant, c.1213+5G>T [IVS6+5G>T], detected in homozygosity. In general, homozygous or compound heterozygous mutations in the ARSB gene, underlie MPS type VI or Maroteaux-Lamy syndrome. Still, even though the novel c.1213+5G>T variant was easy to detect by both NGS and Sanger sequencing, only through indirect studies and functional analyses could we present proof of principle on its pathogenicity. Globally, this case reminds us that whenever a novel variant is detected, its pathogenicity must be carefully assessed before a definitive diagnosis is established, while highlighting alternative approaches that may be used to assess its effect in the absence RNA/cDNA sample(s) from the proband. This is particularly relevant for intronic variants such as the one here reported. Special attention will be given to the use of reporter minigene systems, which may be constructed/designed to dissect the effect of this sort of alterations, providing an insight into their consequences over the normal pre-mRNA splicing process of the affected gene.This work was partially supported by the Fundação Millennium bcp (bcp/LIM/DGH/2014), N2020 (NORTE‐01‐0246‐FEDER‐000014) and Fundação para a Ciência e Tecnologia (FCT) IP (project: PTDC/SAU‐ GMG/117046/2010). M.F.C. and J.I.S. were grantees from the FCT (SFRH/BPD/101965/2014; SFRH/BD/124372/2016).MDPIRepositório Científico do Instituto Nacional de SaúdeCoutinho, Maria FranciscaEncarnação, MarisaMatos, LilianaSilva, LisbethRibeiro, DiogoSantos, Juliana InêsPrata, Maria JoãoVilarinho, LauraAlves, Sandra2020-04-20T10:43:29Z2020-01-212020-01-21T00:00:00Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleapplication/pdfhttp://hdl.handle.net/10400.18/6457engDiagnostics (Basel). 2020 Jan 21;10(2):E58. doi: 10.3390/diagnostics100200582075-441810.3390/diagnostics10020058info:eu-repo/semantics/openAccessreponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãoinstacron:RCAAP2023-07-20T15:41:44Zoai:repositorio.insa.pt:10400.18/6457Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-19T18:41:36.408413Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãofalse |
dc.title.none.fl_str_mv |
Molecular characterization of a novel splicing mutation underlying mucopolysaccharidosis (MPS) type VI-Indirect proof of principle on its pathogenicity |
title |
Molecular characterization of a novel splicing mutation underlying mucopolysaccharidosis (MPS) type VI-Indirect proof of principle on its pathogenicity |
spellingShingle |
Molecular characterization of a novel splicing mutation underlying mucopolysaccharidosis (MPS) type VI-Indirect proof of principle on its pathogenicity Coutinho, Maria Francisca Lysosomal Storage Disorders Functional Studies Mucopolysaccharidosis Type VI Next-generation Sequencing MPS VI Gene ARSB Doenças Genéticas Doenças Lisossomais de Sobrecarga Mutações de Splicing Mucopolissacaridoses |
title_short |
Molecular characterization of a novel splicing mutation underlying mucopolysaccharidosis (MPS) type VI-Indirect proof of principle on its pathogenicity |
title_full |
Molecular characterization of a novel splicing mutation underlying mucopolysaccharidosis (MPS) type VI-Indirect proof of principle on its pathogenicity |
title_fullStr |
Molecular characterization of a novel splicing mutation underlying mucopolysaccharidosis (MPS) type VI-Indirect proof of principle on its pathogenicity |
title_full_unstemmed |
Molecular characterization of a novel splicing mutation underlying mucopolysaccharidosis (MPS) type VI-Indirect proof of principle on its pathogenicity |
title_sort |
Molecular characterization of a novel splicing mutation underlying mucopolysaccharidosis (MPS) type VI-Indirect proof of principle on its pathogenicity |
author |
Coutinho, Maria Francisca |
author_facet |
Coutinho, Maria Francisca Encarnação, Marisa Matos, Liliana Silva, Lisbeth Ribeiro, Diogo Santos, Juliana Inês Prata, Maria João Vilarinho, Laura Alves, Sandra |
author_role |
author |
author2 |
Encarnação, Marisa Matos, Liliana Silva, Lisbeth Ribeiro, Diogo Santos, Juliana Inês Prata, Maria João Vilarinho, Laura Alves, Sandra |
author2_role |
author author author author author author author author |
dc.contributor.none.fl_str_mv |
Repositório Científico do Instituto Nacional de Saúde |
dc.contributor.author.fl_str_mv |
Coutinho, Maria Francisca Encarnação, Marisa Matos, Liliana Silva, Lisbeth Ribeiro, Diogo Santos, Juliana Inês Prata, Maria João Vilarinho, Laura Alves, Sandra |
dc.subject.por.fl_str_mv |
Lysosomal Storage Disorders Functional Studies Mucopolysaccharidosis Type VI Next-generation Sequencing MPS VI Gene ARSB Doenças Genéticas Doenças Lisossomais de Sobrecarga Mutações de Splicing Mucopolissacaridoses |
topic |
Lysosomal Storage Disorders Functional Studies Mucopolysaccharidosis Type VI Next-generation Sequencing MPS VI Gene ARSB Doenças Genéticas Doenças Lisossomais de Sobrecarga Mutações de Splicing Mucopolissacaridoses |
description |
Here, we present the molecular diagnosis of a patient with a general clinical suspicion of Mucopolysaccharidosis, highlighting the different tools used to perform its molecular characterization. In order to decrease the turnaround time for the final report and contribute to reduce the "diagnostic odyssey", which frequently afflicts affected families, the proband's sample was simultaneously screened for mutations in a number of lysosomal function-related genes with targeted next-generation sequencing (NGS) protocol. After variant calling, the most probable cause for disease was a novel ARSB intronic variant, c.1213+5G>T [IVS6+5G>T], detected in homozygosity. In general, homozygous or compound heterozygous mutations in the ARSB gene, underlie MPS type VI or Maroteaux-Lamy syndrome. Still, even though the novel c.1213+5G>T variant was easy to detect by both NGS and Sanger sequencing, only through indirect studies and functional analyses could we present proof of principle on its pathogenicity. Globally, this case reminds us that whenever a novel variant is detected, its pathogenicity must be carefully assessed before a definitive diagnosis is established, while highlighting alternative approaches that may be used to assess its effect in the absence RNA/cDNA sample(s) from the proband. This is particularly relevant for intronic variants such as the one here reported. Special attention will be given to the use of reporter minigene systems, which may be constructed/designed to dissect the effect of this sort of alterations, providing an insight into their consequences over the normal pre-mRNA splicing process of the affected gene. |
publishDate |
2020 |
dc.date.none.fl_str_mv |
2020-04-20T10:43:29Z 2020-01-21 2020-01-21T00:00:00Z |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article |
format |
article |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
http://hdl.handle.net/10400.18/6457 |
url |
http://hdl.handle.net/10400.18/6457 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
Diagnostics (Basel). 2020 Jan 21;10(2):E58. doi: 10.3390/diagnostics10020058 2075-4418 10.3390/diagnostics10020058 |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
application/pdf |
dc.publisher.none.fl_str_mv |
MDPI |
publisher.none.fl_str_mv |
MDPI |
dc.source.none.fl_str_mv |
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Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação |
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RCAAP |
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Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
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Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
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Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação |
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