Screening of genes related to inorganic phosphate in families with primary brain calcifications (PBC)
Ano de defesa: | 2016 |
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Autor(a) principal: | |
Orientador(a): | |
Banca de defesa: | |
Tipo de documento: | Tese |
Tipo de acesso: | Acesso aberto |
Idioma: | eng |
Instituição de defesa: |
Universidade Federal de Pernambuco
|
Programa de Pós-Graduação: |
Programa de Pos Graduacao em Ciencias Biologicas
|
Departamento: |
Não Informado pela instituição
|
País: |
Brasil
|
Palavras-chave em Português: | |
Link de acesso: | https://repositorio.ufpe.br/handle/123456789/26882 |
Resumo: | Primary brain calcification (PBC), also known as idiopathic brain calcification or Fahr's disease, is a rare neurological condition that is characterized by calcium phosphate deposits in the basal ganglia and adjacent areas, movement disorders, headache and neuropsychiatric symptoms. It presents autosomic dominant inheritance and it is associated with two inorganic phosphate transporter coding genes: SLC20A2 and XPR1. Two other genes related to the blood-brain barrier maintenance and integrity are also linked to PBC, the platelet-derived growth factor-β and its receptor (PDGFB and PDGFRB), although their roles in the formation mechanism of the calcifications is not clear yet. For this study, besides the four genes above mentioned, other members of the platelet-derived grown factor family (PDGFA, PDGFRA, PDGFC and PDGFD) have also been selected as candidate genes, for which new primer pairs were designed. All genes above were screened for new variants by Sanger sequencing in fifteen Brazilian unrelated patients with brain calcifications. Sequence in silico analysis was performed using CLC Main Workbench 6.9 software and online tools available in NCBI and GOLDENPATH platforms, resulting in the identification of the first de novo SLC20A2 mutation in a patient diagnosed with PBC (NM_006749.4:c.1158C>G; NP_006740.1:p.Y386*). SLC20A2 is to-date the main gene associated with PBC, with affecting-variants observed in ~50% cases. In order to find SLC20A2 deletions and/or duplications not detected by sequencing, all Brazilian probands were screened by QMPSF (Quantitative Multiplex PCR of Short fluorescent Fragments) and a duplication of the terminal exon was found in a patient with brain calcifications and hyperparatiroidism. Simultaneously, twenty-four French unrelated patients with PBC were also analyzed by QMPSF and partial SLC20A2 deletions were detected in four patients: two with deletion of the exon 2, where the start codon is located; one with deletion of the exon 4; and one with deletion of exons 4 and 5. These results reinforce SLC20A2 role as the main gene associated to PBC, as well as demonstrate that copy number variation analyses, even when revealing only partial deletions or duplications of a gene, are complementary to sequencing and work side by side in the search of genetic variations involved in this disease. |
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FERREIRA, Joana Braga de Moraes Marqueshttp://lattes.cnpq.br/3595342387709218http://lattes.cnpq.br/6651024132073862OLIVEIRA, João Ricardo Mendes de2018-09-24T17:34:42Z2018-09-24T17:34:42Z2016-03-10https://repositorio.ufpe.br/handle/123456789/26882Primary brain calcification (PBC), also known as idiopathic brain calcification or Fahr's disease, is a rare neurological condition that is characterized by calcium phosphate deposits in the basal ganglia and adjacent areas, movement disorders, headache and neuropsychiatric symptoms. It presents autosomic dominant inheritance and it is associated with two inorganic phosphate transporter coding genes: SLC20A2 and XPR1. Two other genes related to the blood-brain barrier maintenance and integrity are also linked to PBC, the platelet-derived growth factor-β and its receptor (PDGFB and PDGFRB), although their roles in the formation mechanism of the calcifications is not clear yet. For this study, besides the four genes above mentioned, other members of the platelet-derived grown factor family (PDGFA, PDGFRA, PDGFC and PDGFD) have also been selected as candidate genes, for which new primer pairs were designed. All genes above were screened for new variants by Sanger sequencing in fifteen Brazilian unrelated patients with brain calcifications. Sequence in silico analysis was performed using CLC Main Workbench 6.9 software and online tools available in NCBI and GOLDENPATH platforms, resulting in the identification of the first de novo SLC20A2 mutation in a patient diagnosed with PBC (NM_006749.4:c.1158C>G; NP_006740.1:p.Y386*). SLC20A2 is to-date the main gene associated with PBC, with affecting-variants observed in ~50% cases. In order to find SLC20A2 deletions and/or duplications not detected by sequencing, all Brazilian probands were screened by QMPSF (Quantitative Multiplex PCR of Short fluorescent Fragments) and a duplication of the terminal exon was found in a patient with brain calcifications and hyperparatiroidism. Simultaneously, twenty-four French unrelated patients with PBC were also analyzed by QMPSF and partial SLC20A2 deletions were detected in four patients: two with deletion of the exon 2, where the start codon is located; one with deletion of the exon 4; and one with deletion of exons 4 and 5. These results reinforce SLC20A2 role as the main gene associated to PBC, as well as demonstrate that copy number variation analyses, even when revealing only partial deletions or duplications of a gene, are complementary to sequencing and work side by side in the search of genetic variations involved in this disease.FACEPEIntrodução: A calcificação cerebral primária (CCP), também conhecida como calcificação idiopática dos núcleos da base ou doença de Fahr, é uma condição neurológica caracterizada por depósitos de fosfato de cálcio dos núcleos da base e região de entorno, parkinsonismo e sintomas neuropsiquiátricos. Apresenta herança autossômica dominante e é associada a dois genes codificantes de transportadores de fosfato inorgânico: o SLC20A2 e o XPR1. Dois outros genes relacionados à manutenção e à integridade da barreira hemato-encefálica, o fator de crescimento plaquetário B e seu receptor (PDGFB e PDGFRB), também foram associados à CCP, embora seus papeis no mecanismo de formação das calcificações ainda não estejam claros. Materiais e Métodos: Além dos quatro genes acima, foram selecionados como candidatos outros genes da família dos fatores de crescimento plaquetário (PDGFA, PDGFRA, PDGFC e PDGFD) e das protocaderinas (PCDH12), para os quais foram confeccionados pares de primers utilizados no seu sequenciamento e para análise de variação de número de cópia. Resultados e Discussão: Quinze famílias brasileiras com CCP foram triadas para novas variantes nos genes candidatos por sequenciamento. A análise in silico do sequenciamento foi feita através do software CLC Combined Workbench versão 6.9 e das ferramentas disponíveis nas plataformas online do NCBI e do GOLDENPATH. A partir dessa análise, foi identificada em um probando a primeira mutação de novo do SLC20A2, o principal gene associado a CCP (NM_006749.4:c.1158C>G; NP_006740.1:p.Y386*). A fim de encontrar deleções e/ou duplicações do SLC20A2 não detectadas por sequenciamento, todos os probandos brasileiros com calcificações cerebrais foram triados através da técnica de QMPSF (do inglês, Quantitative Multiplex PCR of Short fluorescent Fragments). Foi encontrada uma duplicação do exon terminal do mesmo gene em um paciente brasileiro com calcificações cerebrais e hiperparatireoidismo. Simultaneamente, foram identificadas deleções parciais no mesmo gene em quatro famílias francesas com CCP. Conclusões: Esses resultados reafirmam o SLC20A2 como o principal gene associado a CCP, bem como demonstram que análises de variação de número de cópia (CNV), ainda que parciais, são complementares ao sequenciamento na busca por variantes genéticas relacionadas a esta doença.engUniversidade Federal de PernambucoPrograma de Pos Graduacao em Ciencias BiologicasUFPEBrasilAttribution-NonCommercial-NoDerivs 3.0 Brazilhttp://creativecommons.org/licenses/by-nc-nd/3.0/br/info:eu-repo/semantics/openAccessCérebro- doençasGenesMetabolismoScreening of genes related to inorganic phosphate in families with primary brain calcifications (PBC)Triagem de genes relacionados ao metabolismo do fosfato inorgânico em famílias com calcificação idiopática dos núcleos da base do cérebro (IBGC)info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/doctoralThesisdoutoradoreponame:Repositório Institucional da UFPEinstname:Universidade Federal de Pernambuco (UFPE)instacron:UFPETHUMBNAILTESE Joana Braga de Moraes Marques Ferreira.pdf.jpgTESE Joana Braga de Moraes Marques Ferreira.pdf.jpgGenerated Thumbnailimage/jpeg1357https://repositorio.ufpe.br/bitstream/123456789/26882/5/TESE%20Joana%20Braga%20de%20Moraes%20Marques%20Ferreira.pdf.jpg5681b22e94d96bdcb8f5d0d768086a6fMD55ORIGINALTESE Joana Braga de Moraes Marques Ferreira.pdfTESE Joana Braga de Moraes Marques Ferreira.pdfapplication/pdf5193351https://repositorio.ufpe.br/bitstream/123456789/26882/1/TESE%20Joana%20Braga%20de%20Moraes%20Marques%20Ferreira.pdf0e24ea46d41e7220625b8f65daf73891MD51CC-LICENSElicense_rdflicense_rdfapplication/rdf+xml; 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dc.title.pt_BR.fl_str_mv |
Screening of genes related to inorganic phosphate in families with primary brain calcifications (PBC) |
dc.title.alternative.pt_BR.fl_str_mv |
Triagem de genes relacionados ao metabolismo do fosfato inorgânico em famílias com calcificação idiopática dos núcleos da base do cérebro (IBGC) |
title |
Screening of genes related to inorganic phosphate in families with primary brain calcifications (PBC) |
spellingShingle |
Screening of genes related to inorganic phosphate in families with primary brain calcifications (PBC) FERREIRA, Joana Braga de Moraes Marques Cérebro- doenças Genes Metabolismo |
title_short |
Screening of genes related to inorganic phosphate in families with primary brain calcifications (PBC) |
title_full |
Screening of genes related to inorganic phosphate in families with primary brain calcifications (PBC) |
title_fullStr |
Screening of genes related to inorganic phosphate in families with primary brain calcifications (PBC) |
title_full_unstemmed |
Screening of genes related to inorganic phosphate in families with primary brain calcifications (PBC) |
title_sort |
Screening of genes related to inorganic phosphate in families with primary brain calcifications (PBC) |
author |
FERREIRA, Joana Braga de Moraes Marques |
author_facet |
FERREIRA, Joana Braga de Moraes Marques |
author_role |
author |
dc.contributor.authorLattes.pt_BR.fl_str_mv |
http://lattes.cnpq.br/3595342387709218 |
dc.contributor.advisorLattes.pt_BR.fl_str_mv |
http://lattes.cnpq.br/6651024132073862 |
dc.contributor.author.fl_str_mv |
FERREIRA, Joana Braga de Moraes Marques |
dc.contributor.advisor1.fl_str_mv |
OLIVEIRA, João Ricardo Mendes de |
contributor_str_mv |
OLIVEIRA, João Ricardo Mendes de |
dc.subject.por.fl_str_mv |
Cérebro- doenças Genes Metabolismo |
topic |
Cérebro- doenças Genes Metabolismo |
description |
Primary brain calcification (PBC), also known as idiopathic brain calcification or Fahr's disease, is a rare neurological condition that is characterized by calcium phosphate deposits in the basal ganglia and adjacent areas, movement disorders, headache and neuropsychiatric symptoms. It presents autosomic dominant inheritance and it is associated with two inorganic phosphate transporter coding genes: SLC20A2 and XPR1. Two other genes related to the blood-brain barrier maintenance and integrity are also linked to PBC, the platelet-derived growth factor-β and its receptor (PDGFB and PDGFRB), although their roles in the formation mechanism of the calcifications is not clear yet. For this study, besides the four genes above mentioned, other members of the platelet-derived grown factor family (PDGFA, PDGFRA, PDGFC and PDGFD) have also been selected as candidate genes, for which new primer pairs were designed. All genes above were screened for new variants by Sanger sequencing in fifteen Brazilian unrelated patients with brain calcifications. Sequence in silico analysis was performed using CLC Main Workbench 6.9 software and online tools available in NCBI and GOLDENPATH platforms, resulting in the identification of the first de novo SLC20A2 mutation in a patient diagnosed with PBC (NM_006749.4:c.1158C>G; NP_006740.1:p.Y386*). SLC20A2 is to-date the main gene associated with PBC, with affecting-variants observed in ~50% cases. In order to find SLC20A2 deletions and/or duplications not detected by sequencing, all Brazilian probands were screened by QMPSF (Quantitative Multiplex PCR of Short fluorescent Fragments) and a duplication of the terminal exon was found in a patient with brain calcifications and hyperparatiroidism. Simultaneously, twenty-four French unrelated patients with PBC were also analyzed by QMPSF and partial SLC20A2 deletions were detected in four patients: two with deletion of the exon 2, where the start codon is located; one with deletion of the exon 4; and one with deletion of exons 4 and 5. These results reinforce SLC20A2 role as the main gene associated to PBC, as well as demonstrate that copy number variation analyses, even when revealing only partial deletions or duplications of a gene, are complementary to sequencing and work side by side in the search of genetic variations involved in this disease. |
publishDate |
2016 |
dc.date.issued.fl_str_mv |
2016-03-10 |
dc.date.accessioned.fl_str_mv |
2018-09-24T17:34:42Z |
dc.date.available.fl_str_mv |
2018-09-24T17:34:42Z |
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.uri.fl_str_mv |
https://repositorio.ufpe.br/handle/123456789/26882 |
url |
https://repositorio.ufpe.br/handle/123456789/26882 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.rights.driver.fl_str_mv |
Attribution-NonCommercial-NoDerivs 3.0 Brazil http://creativecommons.org/licenses/by-nc-nd/3.0/br/ info:eu-repo/semantics/openAccess |
rights_invalid_str_mv |
Attribution-NonCommercial-NoDerivs 3.0 Brazil http://creativecommons.org/licenses/by-nc-nd/3.0/br/ |
eu_rights_str_mv |
openAccess |
dc.publisher.none.fl_str_mv |
Universidade Federal de Pernambuco |
dc.publisher.program.fl_str_mv |
Programa de Pos Graduacao em Ciencias Biologicas |
dc.publisher.initials.fl_str_mv |
UFPE |
dc.publisher.country.fl_str_mv |
Brasil |
publisher.none.fl_str_mv |
Universidade Federal de Pernambuco |
dc.source.none.fl_str_mv |
reponame:Repositório Institucional da UFPE instname:Universidade Federal de Pernambuco (UFPE) instacron:UFPE |
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UFPE |
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Repositório Institucional da UFPE |
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