We investigated the mutation spectrum of the TANK-Binding Kinase 1 (TBK1) gene and its associated phenotypic spectrum by exonic resequencing of TBK1 in a cohort of 2,538 patients with frontotemporal dementia (FTD), amyotrophic lateral sclerosis (ALS), or FTD plus ALS, ascertained within the European Early-Onset Dementia Consortium. We assessed pathogenicity of predicted protein-truncating mutations by measuring loss of RNA expression. Functional effect of in-frame amino acid deletions and missense mutations was further explored in vivo on protein level and in vitro by an NFκB-induced luciferase reporter assay and measuring phosphorylated TBK1. The protein-truncating mutations led to the loss of transcript through nonsense-mediated mRNA decay. For the in-frame amino acid deletions, we demonstrated loss of TBK1 or phosphorylated TBK1 protein. An important fraction of the missense mutations compromised NFκB activation indicating that at least some functions of TBK1 are lost. Although missense mutations were also present in controls, over three times more mutations affecting TBK1 functioning were found in the mutation fraction observed in patients only, suggesting high-risk alleles (P = 0.03). Total mutation frequency for confirmed TBK1 LoF mutations in the European cohort was 0.7%, with frequencies in the clinical subgroups of 0.4% in FTD, 1.3% in ALS, and 3.6% in FTD-ALS.

Antwerp University Hospital Edegem

Center for Molecular Neurology VIB Antwerp Belgium

Center for Networker Biomedical Research in Neurodegenerative Diseases Madrid Spain

CIBERNED Instituto de Salud Carlos 3 Madrid Spain

Clinical Neuroscience Unit Department of Neurology University of Bonn Bonn Germany

Deparment of Neurology Complejo Asistencial Universitario de Palencia Palencia Spain

Department of Biochemistry Molecular Medicine Center Medical University Sofia Sofia Bulgaria

Department of Cognitive Science and Psychology New Bulgarian University Sofia Bulgaria

Department of Geriatric Medicine Genetics unit Karolinska University Hospital Stockholm Sweden

Department of Neurobiology Care Sciences and Society KI Alzheimer Disease Research Center Karolinska Institutet Stockholm Sweden

Department of Neurodegeneration Hertie Institute for Clinical Brain Research and Centre of Neurology Tübingen Germany

Department of Neurology and Memory Clinic Hospital Network Antwerp Middelheim and Hoge Beuken Antwerp Belgium

Department of Neurology Antwerp University Hospital Edegem Belgium

Department of Neurology Fundación Jiménez Díaz Madrid Spain

Department of Neurology IIB Sant Pau Hospital de la Santa Creu i Sant Pau Universitat Autònoma de Barcelona Barcelona Spain

Department of Neurology Ludwig Maximilians Universität München Munich Germany

Department of Neurology Medical University Sofia Sofia Bulgaria

Department of Neurology University Hospital Ghent and University of Ghent Ghent Belgium

Department of Neurology University Hospitals Leuven Leuven Belgium

Department of Neurosciences Faculty of Medicine KU Leuven Leuven Belgium

Department of Neurosciences Psychology Drug Research and Child Health University of Florence Florence Italy

Department of Psychiatry and Psychotherapy Technische Universität München München Germany

Department of Psychiatry and Psychotherapy University of Bonn Bonn Germany

Department of Psychiatry and Psychotherapy University of Cologne Cologne Germany

Faculty of Medicine and Institute of Molecular Medicine University of Lisbon Lisbon Portugal

Fundació ACE Institut Català de Neurociències Aplicades Barcelona Spain

General Hospital Glorieux Ronse

General Hospital Sint Jan Brugge Bruges

General Hospital Sint Maria Halle

German Center for Neurodegenerative Diseases Bonn Germany

German Center for Neurodegenerative Diseases Munich Germany

German Research Center for Neurodegenerative Diseases Tübingen Germany

Hertie Institute for Clinical Brain Research Tübingen Germany

Hôpitaux Universitaires de Genève et Université de Genève Genève Switzerland and IRCCS Fatebenefratelli Brescia Italy

Hospital Clínic IDIBAPS Barcelona Spain

Hospital Network Antwerp Antwerp

Hospital Santa Maria Lisbon Portugal

Institute Born Bunge University of Antwerp Antwerp Belgium

Institute of Human Genetics University of Bonn Bonn Germany

IRCCS Don Carlo Gnocchi Scandicci Florence Italy

Jessa Hospital Hasselt

MAC Memory Center and Molecular Markers Laboratory IRCCS Istituto Centro San Giovanni di Dio Fatebenefratelli Brescia Italy

Medical University of Vienna Vienna Austria

Molecular Markers Laboratory IRCCS Istituto Centro San Giovanni di Dio Fatebenefratelli Brescia Italy

Neuroepidemiology and Ageing Research Unit School of Public Health The Imperial College of Science Technology and Medicine London UK

Neurological Tissue Bank of the Biobanc Hospital Clinic Institut d'Investigacions Biomediques August Pi i Sunyer Barcelona Spain

Neurology Unit University of Brescia Brescia Italy

NeuroRARE Centre for Rare and Genetic Neurological and Neuromuscular Diseases and Neurogenetics Athens Greece

Saint Luc University Hospital Brussels

Thomayer Hospital Prague and Charles University Prague Czech Republic

University Hospital Brussels Brussels

University Hospital Ghent Ghent

University Hospital Mútua de Terrassa and Fundació Docència i Recerca Mútua Terrassa University of Barcelona School of Medicine Terrassa Barcelona Spain

University Hospitals Leuven Gasthuisberg Leuven

University of Coimbra Coimbra Portugal

University of Liège and Memory Clinic CHU Liège Liège

University of Verona Verona Italy

West London Cognitive Disorders Treatment and Research Unit West London Mental Health Trust London TW8 8DS UK

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