Parkinson's disease (PD) is generally considered a sporadic disorder, but a strong genetic background is often found. The aim of this study was to identify the underlying genetic cause of PD in two affected siblings and to subsequently assess the role of mutations in Cathepsin B (CTSB) in susceptibility to PD. A typical PD family was identified and whole-exome sequencing was performed in two affected siblings. Variants of interest were validated using Sanger sequencing. CTSB p.Gly284Val was genotyped in 2077 PD patients and 615 unrelated healthy controls from the Czech Republic, Ireland, Poland, Ukraine, and the USA. The gene burden analysis was conducted for the CTSB gene in an additional 769 PD probands from Mayo Clinic Florida familial PD cohort. CTSB expression and activity in patient-derived fibroblasts and controls were evaluated by qRT-PCR, western blot, immunocytochemistry, and enzymatic assay. The CTSB p.Gly284Val candidate variant was only identified in affected family members. Functional analysis of CTSB patient-derived fibroblasts under basal conditions did not reveal overt changes in endogenous expression, subcellular localization, or enzymatic activity in the heterozygous carrier of the CTSB variant. The identification of the CTSB p.Gly284Val may support the hypothesis that the CTSB locus harbors variants with differing penetrance that can determine the disease risk.

Applied Neuroscience Research Group Central European Institute of Technology CEITEC MU Masaryk University 601 77 Brno Czech Republic

Bielanski Hospital 01 809 Warsaw Poland

Clinical Department of Neurology Extrapyramidal Disorders and Alzheimer's Outpatient Clinic Central Clinical Hospital of the Ministry of the Interior and Administration in Warsaw 02 507 Warsaw Poland

Department of Clinical Genomics Mayo Clinic Florida Jacksonville FL 32224 USA

Department of Medical Genetics Institute of Mother and Child 01 211 Warsaw Poland

Department of Neurology and Restorative Medicine Health Institute dr Boczarska Jedynak 32 600 Oswiecim Poland

Department of Neurology Faculty of Health Science Medical University of Warsaw 02 091 Warsaw Poland

Department of Neurology Faculty of Medical Sciences in Katowice Medical University of Silesia 40 055 Katowice Poland

Department of Neurology Jagiellonian University Medical College 31 008 Krakow Poland

Department of Neurology Mayo Clinic Florida Jacksonville FL 32224 USA

Department of Neurology St Adalbert Hospital Copernicus PL Ltd 80 462 Gdansk Poland

Department of Neuroscience Mayo Clinic Florida Jacksonville FL 32224 USA

Division of Neurological and Psychiatric Nursing Faculty of Health Sciences Medical University of Gdansk 80 210 Gdansk Poland

Edmond J Safra Program in Parkinson's Disease and the Morton and Gloria Shulman Movement Disorders Clinic Toronto Western Hospital Toronto ON M5T 2S8 Canada

Faculty of Medicine and Health Sciences Andrzej Frycz Modrzewski Krakow University 30 705 Cracow Poland

Institute of Genetics and Biotechnology Faculty of Biology University of Warsaw 00 927 Warsaw Poland

Lviv Regional Clinical Hospital 79010 Lviv Ukraine

Neuroscience PhD Program Mayo Graduate School Mayo Clinic Florida Jacksonville FL 32224 USA

School of Medicine and Medical Science University College Dublin D04 V1W8 Dublin Ireland

St Anne's University Hospital and Faculty of Medicine Masaryk University 601 77 Brno Czech Republic

The Dublin Neurological Institute Mater Misericordiae University Hospital D07 W7XF Dublin Ireland

Uzhhorod National University 88 000 Uzhhorod Ukraine

Uzhhorod Regional Clinical Centre of Neurosurgery and Neurology 88018 Uzhhorod Ukraine

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