Comprehensive molecular-genetic analysis of a hearing loss cohort from Argentina: Functional validation of novel variants identified

Autores
Buonfiglio, Paula Inés; Lotersztein, Vanesa; Menazzi, Sabastián; Plazas, Paola Viviana; Elgoyhen, Ana Belen; Dalamon, Viviana Karina
Año de publicación
2022
Idioma
inglés
Tipo de recurso
documento de conferencia
Estado
versión publicada
Descripción
Hearing loss (HL) is the most prevalent sensorineural deficit, affecting about 20% of the global Population, with nearly half due to genetic causes. About 1 in 500 newborns present congenital HL and more than 100 genes are involved. Most of the patients are non-syndromic with an autosomal recessive mode of inheritance (80%), caused most frequently by GJB2 and GJB6 genes. Inthis work, we aimed to identify the genetic cause of HL designing a multistep approach to analyze target genes.Besides, we performed in silico and in vivo analyses in order to further study some of the identified variants in the zebrafish model. A total of 650 patients were studied by Sanger Sequencing and Gap-PCR in GJB2 and GJB6 genes respectively, diagnosing 15.5% of sporadic cases and 36% of familial ones. Overall, 50 different sequence variants were detected. Next, 50 patients with moderate HL were tested for deletions in STRC gene by MLPA technique. After initial screening, 38 families were selected to be analyzed by Whole Exome Sequencing, achieving Diagnosis in 40%. Half of the identified variants were novel. One of them was a missense variant detected in a familial case in MYO6 gene. To further analyze the functional implication of this variant, a protein modeling with I-Tasser software was performed revealing its pathogenic effect. In order to functional validate this candidate variant a knockdown phenotype rescue assay using anantisense oligonucleotide (morpholino) in zebrafish was carried out demonstrating the deleterious effect on the auditory system. In the present study, we showcased that our algorithm is suitable for the sequential multigenic approach to HL in our cohort. This involves both everyday routine molecular biology and next generation sequencing techniques to identify the genetic etiology ofHL. Furthermore, in silico and in vivo analyses provide not only evidence to validate novel variants but also new knowledge to better understand the genetic basis of HL.
Fil: Buonfiglio, Paula Inés. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; Argentina
Fil: Lotersztein, Vanesa. Ministerio de Defensa. Ejército Argentino. Hospital Militar Central Cirujano Mayor "Dr. Cosme Argerich"; Argentina
Fil: Menazzi, Sabastián. Dirección Nacional de Instituto de Investigación. Administración Nacional de Laboratorios e Institutos de Salud "Dr. Carlos G. Malbrán". Centro Nacional de Genética Médica; Argentina
Fil: Plazas, Paola Viviana. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; Argentina. Universidad de Buenos Aires. Facultad de Medicina. Cátedra de Farmacología. 3º Cátedra de Farmacología; Argentina
Fil: Elgoyhen, Ana Belen. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; Argentina. Universidad de Buenos Aires. Facultad de Medicina. Cátedra de Farmacología. 3º Cátedra de Farmacología; Argentina
Fil: Dalamon, Viviana Karina. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; Argentina
LXVII Reunión anual de la Sociedad Argentina de Investigación Clínica; LXX Reunión anual de la Sociedad Argentina de Inmunología; 3er Congreso franco-argentino de inmunología; Reunión anual 2022 de la Sociedad Argentina de Fisiología
Mar del Plata
Argentina
Sociedad Argentina de Investigacion Clinica
Sociedad Argentina de Inmunología
Sociedad Argentina de Fisiología
Materia
HEARING LOSS
NOVEL VARIANTS
ZEBRAFISH
IN-VIVO
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/243014
Acceder
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repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Comprehensive molecular-genetic analysis of a hearing loss cohort from Argentina: Functional validation of novel variants identifiedBuonfiglio, Paula InésLotersztein, VanesaMenazzi, SabastiánPlazas, Paola VivianaElgoyhen, Ana BelenDalamon, Viviana KarinaHEARING LOSSNOVEL VARIANTSZEBRAFISHIN-VIVOhttps://purl.org/becyt/ford/3.1https://purl.org/becyt/ford/3Hearing loss (HL) is the most prevalent sensorineural deficit, affecting about 20% of the global Population, with nearly half due to genetic causes. About 1 in 500 newborns present congenital HL and more than 100 genes are involved. Most of the patients are non-syndromic with an autosomal recessive mode of inheritance (80%), caused most frequently by GJB2 and GJB6 genes. Inthis work, we aimed to identify the genetic cause of HL designing a multistep approach to analyze target genes.Besides, we performed in silico and in vivo analyses in order to further study some of the identified variants in the zebrafish model. A total of 650 patients were studied by Sanger Sequencing and Gap-PCR in GJB2 and GJB6 genes respectively, diagnosing 15.5% of sporadic cases and 36% of familial ones. Overall, 50 different sequence variants were detected. Next, 50 patients with moderate HL were tested for deletions in STRC gene by MLPA technique. After initial screening, 38 families were selected to be analyzed by Whole Exome Sequencing, achieving Diagnosis in 40%. Half of the identified variants were novel. One of them was a missense variant detected in a familial case in MYO6 gene. To further analyze the functional implication of this variant, a protein modeling with I-Tasser software was performed revealing its pathogenic effect. In order to functional validate this candidate variant a knockdown phenotype rescue assay using anantisense oligonucleotide (morpholino) in zebrafish was carried out demonstrating the deleterious effect on the auditory system. In the present study, we showcased that our algorithm is suitable for the sequential multigenic approach to HL in our cohort. This involves both everyday routine molecular biology and next generation sequencing techniques to identify the genetic etiology ofHL. Furthermore, in silico and in vivo analyses provide not only evidence to validate novel variants but also new knowledge to better understand the genetic basis of HL.Fil: Buonfiglio, Paula Inés. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; ArgentinaFil: Lotersztein, Vanesa. Ministerio de Defensa. Ejército Argentino. Hospital Militar Central Cirujano Mayor "Dr. Cosme Argerich"; ArgentinaFil: Menazzi, Sabastián. Dirección Nacional de Instituto de Investigación. Administración Nacional de Laboratorios e Institutos de Salud "Dr. Carlos G. Malbrán". Centro Nacional de Genética Médica; ArgentinaFil: Plazas, Paola Viviana. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; Argentina. Universidad de Buenos Aires. Facultad de Medicina. Cátedra de Farmacología. 3º Cátedra de Farmacología; ArgentinaFil: Elgoyhen, Ana Belen. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; Argentina. Universidad de Buenos Aires. Facultad de Medicina. Cátedra de Farmacología. 3º Cátedra de Farmacología; ArgentinaFil: Dalamon, Viviana Karina. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; ArgentinaLXVII Reunión anual de la Sociedad Argentina de Investigación Clínica; LXX Reunión anual de la Sociedad Argentina de Inmunología; 3er Congreso franco-argentino de inmunología; Reunión anual 2022 de la Sociedad Argentina de FisiologíaMar del PlataArgentinaSociedad Argentina de Investigacion ClinicaSociedad Argentina de InmunologíaSociedad Argentina de FisiologíaFundación Revista Medicina2022info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/conferenceObjectReuniónJournalhttp://purl.org/coar/resource_type/c_5794info:ar-repo/semantics/documentoDeConferenciaapplication/pdfapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/243014Comprehensive molecular-genetic analysis of a hearing loss cohort from Argentina: Functional validation of novel variants identified; LXVII Reunión anual de la Sociedad Argentina de Investigación Clínica; LXX Reunión anual de la Sociedad Argentina de Inmunología; 3er Congreso franco-argentino de inmunología; Reunión anual 2022 de la Sociedad Argentina de Fisiología; Mar del Plata; Argentina; 2022; 68-691669-9106CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/https://www.medicinabuenosaires.com/PMID/36368022.pdfNacionalinfo:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-03T09:56:17Zoai:ri.conicet.gov.ar:11336/243014instacron:CONICETInstitucionalhttp://ri.conicet.gov.ar/Organismo científico-tecnológicoNo correspondehttp://ri.conicet.gov.ar/oai/requestdasensio@conicet.gov.ar; lcarlino@conicet.gov.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:34982025-09-03 09:56:17.352CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Comprehensive molecular-genetic analysis of a hearing loss cohort from Argentina: Functional validation of novel variants identified
title Comprehensive molecular-genetic analysis of a hearing loss cohort from Argentina: Functional validation of novel variants identified
spellingShingle Comprehensive molecular-genetic analysis of a hearing loss cohort from Argentina: Functional validation of novel variants identified
Buonfiglio, Paula Inés
HEARING LOSS
NOVEL VARIANTS
ZEBRAFISH
IN-VIVO
title_short Comprehensive molecular-genetic analysis of a hearing loss cohort from Argentina: Functional validation of novel variants identified
title_full Comprehensive molecular-genetic analysis of a hearing loss cohort from Argentina: Functional validation of novel variants identified
title_fullStr Comprehensive molecular-genetic analysis of a hearing loss cohort from Argentina: Functional validation of novel variants identified
title_full_unstemmed Comprehensive molecular-genetic analysis of a hearing loss cohort from Argentina: Functional validation of novel variants identified
title_sort Comprehensive molecular-genetic analysis of a hearing loss cohort from Argentina: Functional validation of novel variants identified
dc.creator.none.fl_str_mv Buonfiglio, Paula Inés
Lotersztein, Vanesa
Menazzi, Sabastián
Plazas, Paola Viviana
Elgoyhen, Ana Belen
Dalamon, Viviana Karina
author Buonfiglio, Paula Inés
author_facet Buonfiglio, Paula Inés
Lotersztein, Vanesa
Menazzi, Sabastián
Plazas, Paola Viviana
Elgoyhen, Ana Belen
Dalamon, Viviana Karina
author_role author
author2 Lotersztein, Vanesa
Menazzi, Sabastián
Plazas, Paola Viviana
Elgoyhen, Ana Belen
Dalamon, Viviana Karina
author2_role author
author
author
author
author
dc.subject.none.fl_str_mv HEARING LOSS
NOVEL VARIANTS
ZEBRAFISH
IN-VIVO
topic HEARING LOSS
NOVEL VARIANTS
ZEBRAFISH
IN-VIVO
purl_subject.fl_str_mv https://purl.org/becyt/ford/3.1
https://purl.org/becyt/ford/3
dc.description.none.fl_txt_mv Hearing loss (HL) is the most prevalent sensorineural deficit, affecting about 20% of the global Population, with nearly half due to genetic causes. About 1 in 500 newborns present congenital HL and more than 100 genes are involved. Most of the patients are non-syndromic with an autosomal recessive mode of inheritance (80%), caused most frequently by GJB2 and GJB6 genes. Inthis work, we aimed to identify the genetic cause of HL designing a multistep approach to analyze target genes.Besides, we performed in silico and in vivo analyses in order to further study some of the identified variants in the zebrafish model. A total of 650 patients were studied by Sanger Sequencing and Gap-PCR in GJB2 and GJB6 genes respectively, diagnosing 15.5% of sporadic cases and 36% of familial ones. Overall, 50 different sequence variants were detected. Next, 50 patients with moderate HL were tested for deletions in STRC gene by MLPA technique. After initial screening, 38 families were selected to be analyzed by Whole Exome Sequencing, achieving Diagnosis in 40%. Half of the identified variants were novel. One of them was a missense variant detected in a familial case in MYO6 gene. To further analyze the functional implication of this variant, a protein modeling with I-Tasser software was performed revealing its pathogenic effect. In order to functional validate this candidate variant a knockdown phenotype rescue assay using anantisense oligonucleotide (morpholino) in zebrafish was carried out demonstrating the deleterious effect on the auditory system. In the present study, we showcased that our algorithm is suitable for the sequential multigenic approach to HL in our cohort. This involves both everyday routine molecular biology and next generation sequencing techniques to identify the genetic etiology ofHL. Furthermore, in silico and in vivo analyses provide not only evidence to validate novel variants but also new knowledge to better understand the genetic basis of HL.
Fil: Buonfiglio, Paula Inés. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; Argentina
Fil: Lotersztein, Vanesa. Ministerio de Defensa. Ejército Argentino. Hospital Militar Central Cirujano Mayor "Dr. Cosme Argerich"; Argentina
Fil: Menazzi, Sabastián. Dirección Nacional de Instituto de Investigación. Administración Nacional de Laboratorios e Institutos de Salud "Dr. Carlos G. Malbrán". Centro Nacional de Genética Médica; Argentina
Fil: Plazas, Paola Viviana. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; Argentina. Universidad de Buenos Aires. Facultad de Medicina. Cátedra de Farmacología. 3º Cátedra de Farmacología; Argentina
Fil: Elgoyhen, Ana Belen. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; Argentina. Universidad de Buenos Aires. Facultad de Medicina. Cátedra de Farmacología. 3º Cátedra de Farmacología; Argentina
Fil: Dalamon, Viviana Karina. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; Argentina
LXVII Reunión anual de la Sociedad Argentina de Investigación Clínica; LXX Reunión anual de la Sociedad Argentina de Inmunología; 3er Congreso franco-argentino de inmunología; Reunión anual 2022 de la Sociedad Argentina de Fisiología
Mar del Plata
Argentina
Sociedad Argentina de Investigacion Clinica
Sociedad Argentina de Inmunología
Sociedad Argentina de Fisiología
description Hearing loss (HL) is the most prevalent sensorineural deficit, affecting about 20% of the global Population, with nearly half due to genetic causes. About 1 in 500 newborns present congenital HL and more than 100 genes are involved. Most of the patients are non-syndromic with an autosomal recessive mode of inheritance (80%), caused most frequently by GJB2 and GJB6 genes. Inthis work, we aimed to identify the genetic cause of HL designing a multistep approach to analyze target genes.Besides, we performed in silico and in vivo analyses in order to further study some of the identified variants in the zebrafish model. A total of 650 patients were studied by Sanger Sequencing and Gap-PCR in GJB2 and GJB6 genes respectively, diagnosing 15.5% of sporadic cases and 36% of familial ones. Overall, 50 different sequence variants were detected. Next, 50 patients with moderate HL were tested for deletions in STRC gene by MLPA technique. After initial screening, 38 families were selected to be analyzed by Whole Exome Sequencing, achieving Diagnosis in 40%. Half of the identified variants were novel. One of them was a missense variant detected in a familial case in MYO6 gene. To further analyze the functional implication of this variant, a protein modeling with I-Tasser software was performed revealing its pathogenic effect. In order to functional validate this candidate variant a knockdown phenotype rescue assay using anantisense oligonucleotide (morpholino) in zebrafish was carried out demonstrating the deleterious effect on the auditory system. In the present study, we showcased that our algorithm is suitable for the sequential multigenic approach to HL in our cohort. This involves both everyday routine molecular biology and next generation sequencing techniques to identify the genetic etiology ofHL. Furthermore, in silico and in vivo analyses provide not only evidence to validate novel variants but also new knowledge to better understand the genetic basis of HL.
publishDate 2022
dc.date.none.fl_str_mv 2022
dc.type.none.fl_str_mv info:eu-repo/semantics/publishedVersion
info:eu-repo/semantics/conferenceObject
Reunión
Journal
http://purl.org/coar/resource_type/c_5794
info:ar-repo/semantics/documentoDeConferencia
status_str publishedVersion
format conferenceObject
dc.identifier.none.fl_str_mv http://hdl.handle.net/11336/243014
Comprehensive molecular-genetic analysis of a hearing loss cohort from Argentina: Functional validation of novel variants identified; LXVII Reunión anual de la Sociedad Argentina de Investigación Clínica; LXX Reunión anual de la Sociedad Argentina de Inmunología; 3er Congreso franco-argentino de inmunología; Reunión anual 2022 de la Sociedad Argentina de Fisiología; Mar del Plata; Argentina; 2022; 68-69
1669-9106
CONICET Digital
CONICET
url http://hdl.handle.net/11336/243014
identifier_str_mv Comprehensive molecular-genetic analysis of a hearing loss cohort from Argentina: Functional validation of novel variants identified; LXVII Reunión anual de la Sociedad Argentina de Investigación Clínica; LXX Reunión anual de la Sociedad Argentina de Inmunología; 3er Congreso franco-argentino de inmunología; Reunión anual 2022 de la Sociedad Argentina de Fisiología; Mar del Plata; Argentina; 2022; 68-69
1669-9106
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/url/https://www.medicinabuenosaires.com/PMID/36368022.pdf
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
dc.format.none.fl_str_mv application/pdf
application/pdf
application/pdf
application/pdf
dc.coverage.none.fl_str_mv Nacional
dc.publisher.none.fl_str_mv Fundación Revista Medicina
publisher.none.fl_str_mv Fundación Revista Medicina
dc.source.none.fl_str_mv reponame:CONICET Digital (CONICET)
instname:Consejo Nacional de Investigaciones Científicas y Técnicas
reponame_str CONICET Digital (CONICET)
collection CONICET Digital (CONICET)
instname_str Consejo Nacional de Investigaciones Científicas y Técnicas
repository.name.fl_str_mv CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicas
repository.mail.fl_str_mv dasensio@conicet.gov.ar; lcarlino@conicet.gov.ar
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