Bioenergetic and proteolytic defects in fibroblasts from patients with sporadic Parkinson's disease.
BACKGROUND
Parkinson's disease (PD) is a complex disease and the current interest and focus of scientific research is both investigating the variety of causes that underlie PD pathogenesis, and identifying reliable biomarkers to diagnose and monitor the progression of pathology. Investigation on pathogenic mechanisms in peripheral cells, such as fibroblasts derived from patients with sporadic PD and age/gender matched controls, might generate deeper understanding of the deficits affecting dopaminergic neurons and, possibly, new tools applicable to clinical practice.


METHODS
Primary fibroblast cultures were established from skin biopsies. Increased susceptibility to the PD-related toxin rotenone was determined with apoptosis- and necrosis-specific cell death assays. Protein quality control was evaluated assessing the efficiency of the Ubiquitin Proteasome System (UPS) and protein levels of autophagic markers. Changes in cellular bioenergetics were monitored by measuring oxygen consumption and glycolysis-dependent medium acidification. The oxido-reductive status was determined by detecting mitochondrial superoxide production and oxidation levels in proteins and lipids.


RESULTS
PD fibroblasts showed higher vulnerability to necrotic cell death induced by complex I inhibitor rotenone, reduced UPS function and decreased maximal and rotenone-sensitive mitochondrial respiration. No changes in autophagy and redox markers were detected.


CONCLUSIONS
Our study shows that increased susceptibility to rotenone and the presence of proteolytic and bioenergetic deficits that typically sustain the neurodegenerative process of PD can be detected in fibroblasts from idiopathic PD patients. Fibroblasts might therefore represent a powerful and minimally invasive tool to investigate PD pathogenic mechanisms, which might translate into considerable advances in clinical management of the disease.
Laboratory of Functional Neurochemistry, Center for Research in Neurodegenerative Diseases, National Neurological Institute C. Mondino, Via Mondino 2, 27100 Pavia, Italy; Department of Brain and Behavioral Sciences, University of Pavia, University of Pavia, Via Ferrata 9, 27100 Pavia, Italy.
Giulia
G
Ambrosi
Laboratory of Functional Neurochemistry, Center for Research in Neurodegenerative Diseases, National Neurological Institute C. Mondino, Via Mondino 2, 27100 Pavia, Italy.
Cristina
C
Ghezzi
Department of Genetics, Erasmus Medical Centre, Dr. Molewaterplein 50, 3015GE Rotterdam, The Netherlands.
Sara
S
Sepe
Department of Genetics, Erasmus Medical Centre, Dr. Molewaterplein 50, 3015GE Rotterdam, The Netherlands; RiMED Foundation, Palermo, Italy.
Chiara
C
Milanese
Department of Genetics, Erasmus Medical Centre, Dr. Molewaterplein 50, 3015GE Rotterdam, The Netherlands; Facultad de Ciencias Naturales y Matemáticas, Universidad del Rosario, Bogotá, Colombia.
Cesar
C
Payan-Gomez
Department of Genetics, Erasmus Medical Centre, Dr. Molewaterplein 50, 3015GE Rotterdam, The Netherlands.
Cintia R
CR
Bombardieri
Laboratory of Functional Neurochemistry, Center for Research in Neurodegenerative Diseases, National Neurological Institute C. Mondino, Via Mondino 2, 27100 Pavia, Italy.
Marie-Therese
MT
Armentero
Laboratory of Functional Neurochemistry, Center for Research in Neurodegenerative Diseases, National Neurological Institute C. Mondino, Via Mondino 2, 27100 Pavia, Italy.
Roberta
R
Zangaglia
Laboratory of Functional Neurochemistry, Center for Research in Neurodegenerative Diseases, National Neurological Institute C. Mondino, Via Mondino 2, 27100 Pavia, Italy.
Claudio
C
Pacchetti
Department of Genetics, Erasmus Medical Centre, Dr. Molewaterplein 50, 3015GE Rotterdam, The Netherlands. Electronic address: p.g.mastroberardino@erasmusmc.nl.
Pier Giorgio
PG
Mastroberardino
Laboratory of Functional Neurochemistry, Center for Research in Neurodegenerative Diseases, National Neurological Institute C. Mondino, Via Mondino 2, 27100 Pavia, Italy. Electronic address: fabio.blandini@mondino.it.
Fabio
F
Blandini
D000255:Adenosine Triphosphate / Q000378:metabolism; D017209:Apoptosis; D001343:Autophagy; D016022:Case-Control Studies; D004734:Energy Metabolism; D005260:Female; D005347:Fibroblasts / Q000187:drug effects / Q000378:metabolism / Q000473:pathology*; D006706:Homeostasis; D006801:Humans; D008297:Male; D008875:Middle Aged; D008928:Mitochondria / Q000378:metabolism*; D010101:Oxygen Consumption / Q000187:drug effects; D010300:Parkinson Disease / Q000378:metabolism / Q000473:pathology*; D046988:Proteasome Endopeptidase Complex / Q000378:metabolism*; D012402:Rotenone / Q000494:pharmacology; D013481:Superoxides / Q000378:metabolism; D025801:Ubiquitin / Q000378:metabolism*; D014475:Uncoupling Agents / Q000494:pharmacology
D025801:Ubiquitin; D014475:Uncoupling Agents; D012402:Rotenone; D013481:Superoxides; D000255:Adenosine Triphosphate; D046988:Proteasome Endopeptidase Complex
Extracellular Flux Analyzer; Fibroblasts; Parkinson's disease; Rotenone; Ubiquitin Proteasome System