Avanços tecnológicos e variabilidade genética da expansão CGG da região promotora do gene FMR1
Autor(a) principal: | |
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Data de Publicação: | 2016 |
Tipo de documento: | Tese |
Idioma: | por |
Título da fonte: | Repositório Institucional da UFG |
Texto Completo: | http://repositorio.bc.ufg.br/tede/handle/tede/6737 |
Resumo: | X-Fragile Syndrome (FXS) is the leading cause of inherited intellectual disability in the world and the second of genetic etiology, with an estimated prevalence of 1/4000 men and 1/8000 women. The most common molecular mechanism in SXF is due to changes in the expression of the FMR1 gene, located in Xq27.3, due to CGG trinucleotide expansions in the promoter region and subsequent methylation of the gene. In spite of presenting consistent clinical findings, they are not exclusive, and the existence of carriers of alteration in the FMR1 gene without apparent clinical manifestations makes it impossible to diagnose SXF based only on the evaluation. In the present study, a methodological proposal for the molecular diagnosis of X-Fragile Syndrome was developed from the methylation-specific triple amplification of the promoter region of the FMR1 gene combined with capillary electrophoresis. Thirty-four patients with clinical indication of SXF were referred to a laboratory of the public health network. After extraction and quantification of the DNA, the samples were amplified in an optimized protocol and the products submitted to 36cm capillary electrophoresis to verify the amount of CGG repeats and the degree of DNA methylation of each sample. Pre-mutation (3%) and six complete mutations (18%) were detected, all of which revealed a high degree of methylation. Considering the clinical signs commonly presented, the patients were also analyzed for the occurrence of Autism Spectrum Disorder (ASD), which shadowing and overlapping the SXF, verifying that 100% of the individuals with complete mutation presented the phenotype. Thus, it was possible to observe small behavioral differences in the patients analyzed, indicating a lighter clinical picture regarding aspects of social interaction and stereotypies. Thus, the new methodological proposal allows to effectively determine the CGG trinucleotide expansions in FMR1 allowing an assertive diagnosis of SXF for the families of patients attended in the public health network in Goiás. |
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Cruz, Aparecido Divino dahttp://lattes.cnpq.br/7868817504129985Pereira, Rinaldo Wellersonhttp://lattes.cnpq.br/9065501029560884Cruz , Aparecido Divino daAyres, Flavio MonteiroOliveira , Kátia Karina Verolli deSilva, Claudio Carlos daReis, Angela Adamski da Silvahttp://lattes.cnpq.br/5243589379211142Gigonzac, Marc Alexandre Duarte2017-01-16T10:58:58Z2016-02-02GIGONZAC, Marc Alexandre Duarte. Avanços tecnológicos e variabilidade genética da expansão CGG da região promotora do gene FMR1. 2016. 70 f . Tese (Doutorado em Biotecnologia e Biodiversidade em Rede Pró-Centro-Oeste) - Universidade Federal de Goiás, Goiânia, 2016.http://repositorio.bc.ufg.br/tede/handle/tede/6737ark:/38995/001300000f22mX-Fragile Syndrome (FXS) is the leading cause of inherited intellectual disability in the world and the second of genetic etiology, with an estimated prevalence of 1/4000 men and 1/8000 women. The most common molecular mechanism in SXF is due to changes in the expression of the FMR1 gene, located in Xq27.3, due to CGG trinucleotide expansions in the promoter region and subsequent methylation of the gene. In spite of presenting consistent clinical findings, they are not exclusive, and the existence of carriers of alteration in the FMR1 gene without apparent clinical manifestations makes it impossible to diagnose SXF based only on the evaluation. In the present study, a methodological proposal for the molecular diagnosis of X-Fragile Syndrome was developed from the methylation-specific triple amplification of the promoter region of the FMR1 gene combined with capillary electrophoresis. Thirty-four patients with clinical indication of SXF were referred to a laboratory of the public health network. After extraction and quantification of the DNA, the samples were amplified in an optimized protocol and the products submitted to 36cm capillary electrophoresis to verify the amount of CGG repeats and the degree of DNA methylation of each sample. Pre-mutation (3%) and six complete mutations (18%) were detected, all of which revealed a high degree of methylation. Considering the clinical signs commonly presented, the patients were also analyzed for the occurrence of Autism Spectrum Disorder (ASD), which shadowing and overlapping the SXF, verifying that 100% of the individuals with complete mutation presented the phenotype. Thus, it was possible to observe small behavioral differences in the patients analyzed, indicating a lighter clinical picture regarding aspects of social interaction and stereotypies. Thus, the new methodological proposal allows to effectively determine the CGG trinucleotide expansions in FMR1 allowing an assertive diagnosis of SXF for the families of patients attended in the public health network in Goiás.A Síndrome do X-Frágil (SXF) é a principal causa de deficiência intelectual herdável no mundo e a segunda de etiologia genética, com uma prevalência estimada de 1/4000 homens e 1/8000 mulheres. O mecanismo molecular mais comum na SXF é decorrente de alterações na expressão do gene FMR1, localizado em Xq27.3, devido a expansões trinucleotídicas CGG na região promotora e subsequente metilação do gene. Apesar de apresentar achados clínicos consistentes, os mesmos não são exclusivos, e a existência de portadores de alteração no gene FMR1 sem manifestações clínicas aparentes impossibilitam o diagnóstico da SXF baseado apenas no exame físico. No presente estudo foi desenvolvido uma proposta metodológica para o diagnóstico molecular da Síndrome do X-Frágil a partir da amplificação tripla metilação-específica da região promotora do gene FMR1 combinada a eletroforese em capilar. Foram utilizados 34 pacientes com indicação clínica de SXF encaminhados para um laboratório da rede pública de saúde. Após extração e quantificação do DNA, as amostras foram amplificadas em protocolo otimizado e os produtos submetidos a eletroforese em capilar de 36cm para verificar a quantidade de repetições CGG e o grau de metilação do DNA de cada amostra. Foram detectadas uma pré-mutação (3%) e seis mutações completas (18%), sendo que todas estas últimas revelaram um alto grau de metilação. Considerando os sinais clínicos comumente apresentados, os pacientes foram também analisados para a ocorrência do Transtorno do Espectro do Autismo (TEA), que sombreia e se sobrepõe à SXF, verificando que 100% dos indivíduos com mutação completa apresentaram o fenótipo. Foi possível observar pequenas diferenças comportamentais nos pacientes analisados, indicando um quadro clínico mais leve quanto aos aspectos da interação social e das estereotipias. Sendo assim, a nova proposta metodológica permite determinar de forma eficaz as expansões trinucleotídicas CGG no FMR1 permitindo um diagnóstico assertivo da SXF para as famílias de pacientes atendidos na rede pública de saúde em Goiás.Submitted by Cássia Santos (cassia.bcufg@gmail.com) on 2017-01-13T10:53:51Z No. of bitstreams: 2 Tese - Marc Alexandre Duarte Gigonzac - 2016.pdf: 12763622 bytes, checksum: 3479eadda35402525c2387337a3a0d69 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5)Approved for entry into archive by Luciana Ferreira (lucgeral@gmail.com) on 2017-01-16T10:58:58Z (GMT) No. of bitstreams: 2 Tese - Marc Alexandre Duarte Gigonzac - 2016.pdf: 12763622 bytes, checksum: 3479eadda35402525c2387337a3a0d69 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5)Made available in DSpace on 2017-01-16T10:58:58Z (GMT). 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dc.title.por.fl_str_mv |
Avanços tecnológicos e variabilidade genética da expansão CGG da região promotora do gene FMR1 |
dc.title.alternative.eng.fl_str_mv |
Technological advances and genetic variability of the CGG expansion of the promoter region of the FMR1 gene |
title |
Avanços tecnológicos e variabilidade genética da expansão CGG da região promotora do gene FMR1 |
spellingShingle |
Avanços tecnológicos e variabilidade genética da expansão CGG da região promotora do gene FMR1 Gigonzac, Marc Alexandre Duarte FMR1 Expansões trinucleotídicas Metilação Diagnóstico molecular FMR1 Trinucleotide expansions Methylation Molecular diagnostics CIENCIAS BIOLOGICAS::GENETICA |
title_short |
Avanços tecnológicos e variabilidade genética da expansão CGG da região promotora do gene FMR1 |
title_full |
Avanços tecnológicos e variabilidade genética da expansão CGG da região promotora do gene FMR1 |
title_fullStr |
Avanços tecnológicos e variabilidade genética da expansão CGG da região promotora do gene FMR1 |
title_full_unstemmed |
Avanços tecnológicos e variabilidade genética da expansão CGG da região promotora do gene FMR1 |
title_sort |
Avanços tecnológicos e variabilidade genética da expansão CGG da região promotora do gene FMR1 |
author |
Gigonzac, Marc Alexandre Duarte |
author_facet |
Gigonzac, Marc Alexandre Duarte |
author_role |
author |
dc.contributor.advisor1.fl_str_mv |
Cruz, Aparecido Divino da |
dc.contributor.advisor1Lattes.fl_str_mv |
http://lattes.cnpq.br/7868817504129985 |
dc.contributor.advisor-co1.fl_str_mv |
Pereira, Rinaldo Wellerson |
dc.contributor.advisor-co1Lattes.fl_str_mv |
http://lattes.cnpq.br/9065501029560884 |
dc.contributor.referee1.fl_str_mv |
Cruz , Aparecido Divino da |
dc.contributor.referee2.fl_str_mv |
Ayres, Flavio Monteiro |
dc.contributor.referee3.fl_str_mv |
Oliveira , Kátia Karina Verolli de |
dc.contributor.referee4.fl_str_mv |
Silva, Claudio Carlos da |
dc.contributor.referee5.fl_str_mv |
Reis, Angela Adamski da Silva |
dc.contributor.authorLattes.fl_str_mv |
http://lattes.cnpq.br/5243589379211142 |
dc.contributor.author.fl_str_mv |
Gigonzac, Marc Alexandre Duarte |
contributor_str_mv |
Cruz, Aparecido Divino da Pereira, Rinaldo Wellerson Cruz , Aparecido Divino da Ayres, Flavio Monteiro Oliveira , Kátia Karina Verolli de Silva, Claudio Carlos da Reis, Angela Adamski da Silva |
dc.subject.por.fl_str_mv |
FMR1 Expansões trinucleotídicas Metilação Diagnóstico molecular |
topic |
FMR1 Expansões trinucleotídicas Metilação Diagnóstico molecular FMR1 Trinucleotide expansions Methylation Molecular diagnostics CIENCIAS BIOLOGICAS::GENETICA |
dc.subject.eng.fl_str_mv |
FMR1 Trinucleotide expansions Methylation Molecular diagnostics |
dc.subject.cnpq.fl_str_mv |
CIENCIAS BIOLOGICAS::GENETICA |
description |
X-Fragile Syndrome (FXS) is the leading cause of inherited intellectual disability in the world and the second of genetic etiology, with an estimated prevalence of 1/4000 men and 1/8000 women. The most common molecular mechanism in SXF is due to changes in the expression of the FMR1 gene, located in Xq27.3, due to CGG trinucleotide expansions in the promoter region and subsequent methylation of the gene. In spite of presenting consistent clinical findings, they are not exclusive, and the existence of carriers of alteration in the FMR1 gene without apparent clinical manifestations makes it impossible to diagnose SXF based only on the evaluation. In the present study, a methodological proposal for the molecular diagnosis of X-Fragile Syndrome was developed from the methylation-specific triple amplification of the promoter region of the FMR1 gene combined with capillary electrophoresis. Thirty-four patients with clinical indication of SXF were referred to a laboratory of the public health network. After extraction and quantification of the DNA, the samples were amplified in an optimized protocol and the products submitted to 36cm capillary electrophoresis to verify the amount of CGG repeats and the degree of DNA methylation of each sample. Pre-mutation (3%) and six complete mutations (18%) were detected, all of which revealed a high degree of methylation. Considering the clinical signs commonly presented, the patients were also analyzed for the occurrence of Autism Spectrum Disorder (ASD), which shadowing and overlapping the SXF, verifying that 100% of the individuals with complete mutation presented the phenotype. Thus, it was possible to observe small behavioral differences in the patients analyzed, indicating a lighter clinical picture regarding aspects of social interaction and stereotypies. Thus, the new methodological proposal allows to effectively determine the CGG trinucleotide expansions in FMR1 allowing an assertive diagnosis of SXF for the families of patients attended in the public health network in Goiás. |
publishDate |
2016 |
dc.date.issued.fl_str_mv |
2016-02-02 |
dc.date.accessioned.fl_str_mv |
2017-01-16T10:58:58Z |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/doctoralThesis |
format |
doctoralThesis |
status_str |
publishedVersion |
dc.identifier.citation.fl_str_mv |
GIGONZAC, Marc Alexandre Duarte. Avanços tecnológicos e variabilidade genética da expansão CGG da região promotora do gene FMR1. 2016. 70 f . Tese (Doutorado em Biotecnologia e Biodiversidade em Rede Pró-Centro-Oeste) - Universidade Federal de Goiás, Goiânia, 2016. |
dc.identifier.uri.fl_str_mv |
http://repositorio.bc.ufg.br/tede/handle/tede/6737 |
dc.identifier.dark.fl_str_mv |
ark:/38995/001300000f22m |
identifier_str_mv |
GIGONZAC, Marc Alexandre Duarte. Avanços tecnológicos e variabilidade genética da expansão CGG da região promotora do gene FMR1. 2016. 70 f . Tese (Doutorado em Biotecnologia e Biodiversidade em Rede Pró-Centro-Oeste) - Universidade Federal de Goiás, Goiânia, 2016. ark:/38995/001300000f22m |
url |
http://repositorio.bc.ufg.br/tede/handle/tede/6737 |
dc.language.iso.fl_str_mv |
por |
language |
por |
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dc.relation.confidence.fl_str_mv |
600 600 600 600 |
dc.relation.department.fl_str_mv |
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dc.relation.cnpq.fl_str_mv |
-5518144268585252051 |
dc.relation.sponsorship.fl_str_mv |
-961409807440757778 |
dc.rights.driver.fl_str_mv |
http://creativecommons.org/licenses/by-nc-nd/4.0/ info:eu-repo/semantics/openAccess |
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http://creativecommons.org/licenses/by-nc-nd/4.0/ |
eu_rights_str_mv |
openAccess |
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Universidade Federal de Goiás |
dc.publisher.program.fl_str_mv |
Programa de Pós-graduação em Biotecnologia e Biodiversidade - Rede Pró-Centro-Oeste (PRPG/UnB) |
dc.publisher.initials.fl_str_mv |
UFG |
dc.publisher.country.fl_str_mv |
Brasil |
dc.publisher.department.fl_str_mv |
Pró-Reitoria de Pós-graduação (PRPG) |
publisher.none.fl_str_mv |
Universidade Federal de Goiás |
dc.source.none.fl_str_mv |
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Repositório Institucional da UFG |
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bd3efa91386c1718a7f26a329fdcb468 4afdbb8c545fd630ea7db775da747b2f d41d8cd98f00b204e9800998ecf8427e d41d8cd98f00b204e9800998ecf8427e 3479eadda35402525c2387337a3a0d69 |
bitstream.checksumAlgorithm.fl_str_mv |
MD5 MD5 MD5 MD5 MD5 |
repository.name.fl_str_mv |
Repositório Institucional da UFG - Universidade Federal de Goiás (UFG) |
repository.mail.fl_str_mv |
tasesdissertacoes.bc@ufg.br |
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1811721529608634368 |