Cargando…

Molecular and Functional Characterisation of a Novel Intragenic 12q24.21 Deletion Resulting in MED13L Haploinsufficiency Syndrome

Background and Objectives: Heterozygous pathogenic variants in the MED13L gene cause impaired intellectual development and distinctive facial features with or without cardiac defects (MIM #616789). This complex neurodevelopmental disorder is characterised by various phenotypic features, including pl...

Descripción completa

Detalles Bibliográficos
Autores principales: Siavrienė, Evelina, Petraitytė, Gunda, Mikštienė, Violeta, Maldžienė, Živilė, Sasnauskienė, Aušra, Žitkutė, Vilmantė, Ambrozaitytė, Laima, Rančelis, Tautvydas, Utkus, Algirdas, Kučinskas, Vaidutis, Preikšaitienė, Eglė
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10385642/
https://www.ncbi.nlm.nih.gov/pubmed/37512036
http://dx.doi.org/10.3390/medicina59071225
_version_ 1785081458597036032
author Siavrienė, Evelina
Petraitytė, Gunda
Mikštienė, Violeta
Maldžienė, Živilė
Sasnauskienė, Aušra
Žitkutė, Vilmantė
Ambrozaitytė, Laima
Rančelis, Tautvydas
Utkus, Algirdas
Kučinskas, Vaidutis
Preikšaitienė, Eglė
author_facet Siavrienė, Evelina
Petraitytė, Gunda
Mikštienė, Violeta
Maldžienė, Živilė
Sasnauskienė, Aušra
Žitkutė, Vilmantė
Ambrozaitytė, Laima
Rančelis, Tautvydas
Utkus, Algirdas
Kučinskas, Vaidutis
Preikšaitienė, Eglė
author_sort Siavrienė, Evelina
collection PubMed
description Background and Objectives: Heterozygous pathogenic variants in the MED13L gene cause impaired intellectual development and distinctive facial features with or without cardiac defects (MIM #616789). This complex neurodevelopmental disorder is characterised by various phenotypic features, including plagiocephaly, strabismus, clubfoot, poor speech, and developmental delay. The aim of this study was to evaluate the clinical significance and consequences of a novel heterozygous intragenic MED13L deletion in a proband with clinical features of a MED13L-related disorder through extensive clinical, molecular, and functional characterisation. Materials and Methods: Combined comparative genomic hybridisation and single-nucleotide polymorphism array (SNP-CGH) was used to identify the changes in the proband’s gDNA sequence (DECIPHER #430183). Intragenic MED13L deletion was specified via quantitative polymerase chain reaction (qPCR) and Sanger sequencing of the proband’s cDNA sample. Western blot and bioinformatics analyses were used to investigate the consequences of this copy number variant (CNV) at the protein level. CRISPR-Cas9 technology was used for a MED13L-gene-silencing experiment in a culture of the control individual’s skin fibroblasts. After the MED13L-gene-editing experiment, subsequent functional fibroblast culture analyses were performed. Results: The analysis of the proband’s cDNA sample allowed for specifying the regions of the breakpoints and identifying the heterozygous deletion that spanned exons 3 to 10 of MED13L, which has not been reported previously. In silico, the deletion was predicted to result in a truncated protein NP_056150.1:p.(Val104Glyfs*5), partly altering the Med13_N domain and losing the MedPIWI and Med13_C domains. After MED13L gene editing was performed, reduced cell viability; an accelerated aging process; and inhibition of the RB1, E2F1, and CCNC gene expression were found to exist. Conclusions: Based on these findings, heterozygous intragenic 12q24.21 deletion in the affected individual resulted in MED13L haploinsufficiency due to the premature termination of protein translation, therefore leading to MED13L haploinsufficiency syndrome.
format Online
Article
Text
id pubmed-10385642
institution National Center for Biotechnology Information
language English
publishDate 2023
publisher MDPI
record_format MEDLINE/PubMed
spelling pubmed-103856422023-07-30 Molecular and Functional Characterisation of a Novel Intragenic 12q24.21 Deletion Resulting in MED13L Haploinsufficiency Syndrome Siavrienė, Evelina Petraitytė, Gunda Mikštienė, Violeta Maldžienė, Živilė Sasnauskienė, Aušra Žitkutė, Vilmantė Ambrozaitytė, Laima Rančelis, Tautvydas Utkus, Algirdas Kučinskas, Vaidutis Preikšaitienė, Eglė Medicina (Kaunas) Article Background and Objectives: Heterozygous pathogenic variants in the MED13L gene cause impaired intellectual development and distinctive facial features with or without cardiac defects (MIM #616789). This complex neurodevelopmental disorder is characterised by various phenotypic features, including plagiocephaly, strabismus, clubfoot, poor speech, and developmental delay. The aim of this study was to evaluate the clinical significance and consequences of a novel heterozygous intragenic MED13L deletion in a proband with clinical features of a MED13L-related disorder through extensive clinical, molecular, and functional characterisation. Materials and Methods: Combined comparative genomic hybridisation and single-nucleotide polymorphism array (SNP-CGH) was used to identify the changes in the proband’s gDNA sequence (DECIPHER #430183). Intragenic MED13L deletion was specified via quantitative polymerase chain reaction (qPCR) and Sanger sequencing of the proband’s cDNA sample. Western blot and bioinformatics analyses were used to investigate the consequences of this copy number variant (CNV) at the protein level. CRISPR-Cas9 technology was used for a MED13L-gene-silencing experiment in a culture of the control individual’s skin fibroblasts. After the MED13L-gene-editing experiment, subsequent functional fibroblast culture analyses were performed. Results: The analysis of the proband’s cDNA sample allowed for specifying the regions of the breakpoints and identifying the heterozygous deletion that spanned exons 3 to 10 of MED13L, which has not been reported previously. In silico, the deletion was predicted to result in a truncated protein NP_056150.1:p.(Val104Glyfs*5), partly altering the Med13_N domain and losing the MedPIWI and Med13_C domains. After MED13L gene editing was performed, reduced cell viability; an accelerated aging process; and inhibition of the RB1, E2F1, and CCNC gene expression were found to exist. Conclusions: Based on these findings, heterozygous intragenic 12q24.21 deletion in the affected individual resulted in MED13L haploinsufficiency due to the premature termination of protein translation, therefore leading to MED13L haploinsufficiency syndrome. MDPI 2023-06-29 /pmc/articles/PMC10385642/ /pubmed/37512036 http://dx.doi.org/10.3390/medicina59071225 Text en © 2023 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Siavrienė, Evelina
Petraitytė, Gunda
Mikštienė, Violeta
Maldžienė, Živilė
Sasnauskienė, Aušra
Žitkutė, Vilmantė
Ambrozaitytė, Laima
Rančelis, Tautvydas
Utkus, Algirdas
Kučinskas, Vaidutis
Preikšaitienė, Eglė
Molecular and Functional Characterisation of a Novel Intragenic 12q24.21 Deletion Resulting in MED13L Haploinsufficiency Syndrome
title Molecular and Functional Characterisation of a Novel Intragenic 12q24.21 Deletion Resulting in MED13L Haploinsufficiency Syndrome
title_full Molecular and Functional Characterisation of a Novel Intragenic 12q24.21 Deletion Resulting in MED13L Haploinsufficiency Syndrome
title_fullStr Molecular and Functional Characterisation of a Novel Intragenic 12q24.21 Deletion Resulting in MED13L Haploinsufficiency Syndrome
title_full_unstemmed Molecular and Functional Characterisation of a Novel Intragenic 12q24.21 Deletion Resulting in MED13L Haploinsufficiency Syndrome
title_short Molecular and Functional Characterisation of a Novel Intragenic 12q24.21 Deletion Resulting in MED13L Haploinsufficiency Syndrome
title_sort molecular and functional characterisation of a novel intragenic 12q24.21 deletion resulting in med13l haploinsufficiency syndrome
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10385642/
https://www.ncbi.nlm.nih.gov/pubmed/37512036
http://dx.doi.org/10.3390/medicina59071225
work_keys_str_mv AT siavrieneevelina molecularandfunctionalcharacterisationofanovelintragenic12q2421deletionresultinginmed13lhaploinsufficiencysyndrome
AT petraitytegunda molecularandfunctionalcharacterisationofanovelintragenic12q2421deletionresultinginmed13lhaploinsufficiencysyndrome
AT mikstienevioleta molecularandfunctionalcharacterisationofanovelintragenic12q2421deletionresultinginmed13lhaploinsufficiencysyndrome
AT maldzienezivile molecularandfunctionalcharacterisationofanovelintragenic12q2421deletionresultinginmed13lhaploinsufficiencysyndrome
AT sasnauskieneausra molecularandfunctionalcharacterisationofanovelintragenic12q2421deletionresultinginmed13lhaploinsufficiencysyndrome
AT zitkutevilmante molecularandfunctionalcharacterisationofanovelintragenic12q2421deletionresultinginmed13lhaploinsufficiencysyndrome
AT ambrozaitytelaima molecularandfunctionalcharacterisationofanovelintragenic12q2421deletionresultinginmed13lhaploinsufficiencysyndrome
AT rancelistautvydas molecularandfunctionalcharacterisationofanovelintragenic12q2421deletionresultinginmed13lhaploinsufficiencysyndrome
AT utkusalgirdas molecularandfunctionalcharacterisationofanovelintragenic12q2421deletionresultinginmed13lhaploinsufficiencysyndrome
AT kucinskasvaidutis molecularandfunctionalcharacterisationofanovelintragenic12q2421deletionresultinginmed13lhaploinsufficiencysyndrome
AT preiksaitieneegle molecularandfunctionalcharacterisationofanovelintragenic12q2421deletionresultinginmed13lhaploinsufficiencysyndrome