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TBK1 mutations in Italian patients with amyotrophic lateral sclerosis: genetic and functional characterisation

BACKGROUND: TANK-binding kinase 1 (TBK1) gene has been recently identified as a causative gene of amyotrophic lateral sclerosis (ALS). METHODS: We sequenced the TBK1 gene in a cohort of 154 Italian patients with ALS with unclear genetic aetiology. We subsequently assessed the pathogenic potential of...

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Autores principales: Pozzi, Laura, Valenza, Fabiola, Mosca, Lorena, Dal Mas, Andrea, Domi, Teuta, Romano, Alessandro, Tarlarini, Claudia, Falzone, Yuri Matteo, Tremolizzo, Lucio, Sorarù, Gianni, Cerri, Federica, Ferraro, Pilar M, Basaia, Silvia, Agosta, Federica, Fazio, Raffaella, Comola, Mauro, Comi, Giancarlo, Ferrari, Maurizio, Quattrini, Angelo, Lunetta, Christian, Penco, Silvana, Bonanomi, Dario, Carrera, Paola, Riva, Nilo
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BMJ Publishing Group 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5629935/
https://www.ncbi.nlm.nih.gov/pubmed/28822984
http://dx.doi.org/10.1136/jnnp-2017-316174
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author Pozzi, Laura
Valenza, Fabiola
Mosca, Lorena
Dal Mas, Andrea
Domi, Teuta
Romano, Alessandro
Tarlarini, Claudia
Falzone, Yuri Matteo
Tremolizzo, Lucio
Sorarù, Gianni
Cerri, Federica
Ferraro, Pilar M
Basaia, Silvia
Agosta, Federica
Fazio, Raffaella
Comola, Mauro
Comi, Giancarlo
Ferrari, Maurizio
Quattrini, Angelo
Lunetta, Christian
Penco, Silvana
Bonanomi, Dario
Carrera, Paola
Riva, Nilo
author_facet Pozzi, Laura
Valenza, Fabiola
Mosca, Lorena
Dal Mas, Andrea
Domi, Teuta
Romano, Alessandro
Tarlarini, Claudia
Falzone, Yuri Matteo
Tremolizzo, Lucio
Sorarù, Gianni
Cerri, Federica
Ferraro, Pilar M
Basaia, Silvia
Agosta, Federica
Fazio, Raffaella
Comola, Mauro
Comi, Giancarlo
Ferrari, Maurizio
Quattrini, Angelo
Lunetta, Christian
Penco, Silvana
Bonanomi, Dario
Carrera, Paola
Riva, Nilo
author_sort Pozzi, Laura
collection PubMed
description BACKGROUND: TANK-binding kinase 1 (TBK1) gene has been recently identified as a causative gene of amyotrophic lateral sclerosis (ALS). METHODS: We sequenced the TBK1 gene in a cohort of 154 Italian patients with ALS with unclear genetic aetiology. We subsequently assessed the pathogenic potential of novel identified TBK1 variants using functional in vitro studies: expression, targeting and activity were evaluated in patient-derived fibroblasts and in cells transfected with mutated-TBK1 plasmids. RESULTS: We identified novel genomic TBK1 variants including two loss-of-function (LoF) (p.Leu59Phefs*16 and c.358+5G>A), two missense (p.Asp118Asn and p.Ile397Thr) and one intronic variant (c.1644–5_1644-2delAATA), in addition to two previously reported pathogenetic missense variants (p.Lys291Glu and p.Arg357Gln). Functional studies in patient-derived fibroblasts revealed that the c.358+5G>A causes aberrant pre-mRNA processing leading TBK1 haploinsufficiency. Biochemical studies in cellular models showed that the truncating variant p.Leu59Phefs*16 abolishes TBK1 protein expression, whereas the p.Asp118Asn variant severely impairs TBK1 phosphorylation activity. Conversely, the p.Ile397Thr variant displayed enhanced phosphorylation activity, whose biological relevance is not clear. CONCLUSION: The observed frequency of TBK1 LoF variants was 1.3% (2/154), increasing up to 3.2% (5/154) by taking into account also the functional missense variants that we were able to classify as potentially pathogenic, supporting the relevance of TBK1 in the Italian population with ALS.
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spelling pubmed-56299352017-10-11 TBK1 mutations in Italian patients with amyotrophic lateral sclerosis: genetic and functional characterisation Pozzi, Laura Valenza, Fabiola Mosca, Lorena Dal Mas, Andrea Domi, Teuta Romano, Alessandro Tarlarini, Claudia Falzone, Yuri Matteo Tremolizzo, Lucio Sorarù, Gianni Cerri, Federica Ferraro, Pilar M Basaia, Silvia Agosta, Federica Fazio, Raffaella Comola, Mauro Comi, Giancarlo Ferrari, Maurizio Quattrini, Angelo Lunetta, Christian Penco, Silvana Bonanomi, Dario Carrera, Paola Riva, Nilo J Neurol Neurosurg Psychiatry Neurogenetics BACKGROUND: TANK-binding kinase 1 (TBK1) gene has been recently identified as a causative gene of amyotrophic lateral sclerosis (ALS). METHODS: We sequenced the TBK1 gene in a cohort of 154 Italian patients with ALS with unclear genetic aetiology. We subsequently assessed the pathogenic potential of novel identified TBK1 variants using functional in vitro studies: expression, targeting and activity were evaluated in patient-derived fibroblasts and in cells transfected with mutated-TBK1 plasmids. RESULTS: We identified novel genomic TBK1 variants including two loss-of-function (LoF) (p.Leu59Phefs*16 and c.358+5G>A), two missense (p.Asp118Asn and p.Ile397Thr) and one intronic variant (c.1644–5_1644-2delAATA), in addition to two previously reported pathogenetic missense variants (p.Lys291Glu and p.Arg357Gln). Functional studies in patient-derived fibroblasts revealed that the c.358+5G>A causes aberrant pre-mRNA processing leading TBK1 haploinsufficiency. Biochemical studies in cellular models showed that the truncating variant p.Leu59Phefs*16 abolishes TBK1 protein expression, whereas the p.Asp118Asn variant severely impairs TBK1 phosphorylation activity. Conversely, the p.Ile397Thr variant displayed enhanced phosphorylation activity, whose biological relevance is not clear. CONCLUSION: The observed frequency of TBK1 LoF variants was 1.3% (2/154), increasing up to 3.2% (5/154) by taking into account also the functional missense variants that we were able to classify as potentially pathogenic, supporting the relevance of TBK1 in the Italian population with ALS. BMJ Publishing Group 2017-10 2017-08-19 /pmc/articles/PMC5629935/ /pubmed/28822984 http://dx.doi.org/10.1136/jnnp-2017-316174 Text en © Article author(s) (or their employer(s) unless otherwise stated in the text of the article) 2017. All rights reserved. No commercial use is permitted unless otherwise expressly granted. This is an Open Access article distributed in accordance with the Creative Commons Attribution Non Commercial (CC BY-NC 4.0) license, which permits others to distribute, remix, adapt, build upon this work non-commercially, and license their derivative works on different terms, provided the original work is properly cited and the use is non-commercial. See: http://creativecommons.org/licenses/by-nc/4.0/
spellingShingle Neurogenetics
Pozzi, Laura
Valenza, Fabiola
Mosca, Lorena
Dal Mas, Andrea
Domi, Teuta
Romano, Alessandro
Tarlarini, Claudia
Falzone, Yuri Matteo
Tremolizzo, Lucio
Sorarù, Gianni
Cerri, Federica
Ferraro, Pilar M
Basaia, Silvia
Agosta, Federica
Fazio, Raffaella
Comola, Mauro
Comi, Giancarlo
Ferrari, Maurizio
Quattrini, Angelo
Lunetta, Christian
Penco, Silvana
Bonanomi, Dario
Carrera, Paola
Riva, Nilo
TBK1 mutations in Italian patients with amyotrophic lateral sclerosis: genetic and functional characterisation
title TBK1 mutations in Italian patients with amyotrophic lateral sclerosis: genetic and functional characterisation
title_full TBK1 mutations in Italian patients with amyotrophic lateral sclerosis: genetic and functional characterisation
title_fullStr TBK1 mutations in Italian patients with amyotrophic lateral sclerosis: genetic and functional characterisation
title_full_unstemmed TBK1 mutations in Italian patients with amyotrophic lateral sclerosis: genetic and functional characterisation
title_short TBK1 mutations in Italian patients with amyotrophic lateral sclerosis: genetic and functional characterisation
title_sort tbk1 mutations in italian patients with amyotrophic lateral sclerosis: genetic and functional characterisation
topic Neurogenetics
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5629935/
https://www.ncbi.nlm.nih.gov/pubmed/28822984
http://dx.doi.org/10.1136/jnnp-2017-316174
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