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Stem cell modeling of mitochondrial parkinsonism reveals key functions of OPA1.

Published version
Peer-reviewed

Type

Article

Change log

Authors

Jonikas, Mindaugas 
Madill, Martin 
Mathy, Alexandre 
Zekoll, Theresa 
Zois, Christos E 

Abstract

OBJECTIVE: Defective mitochondrial function attributed to optic atrophy 1 (OPA1) mutations causes primarily optic atrophy and, less commonly, neurodegenerative syndromes. The pathomechanism by which OPA1 mutations trigger diffuse loss of neurons in some, but not all, patients is unknown. Here, we used a tractable induced pluripotent stem cell (iPSC)-based model to capture the biology of OPA1 haploinsufficiency in cases presenting with classic eye disease versus syndromic parkinsonism. METHODS: iPSCs were generated from 2 patients with OPA1 haploinsufficiency and 2 controls and differentiated into dopaminergic neurons. Metabolic profile was determined by extracellular flux analysis, respiratory complex levels using immunoblotting, and complex I activity by a colorimetric assay. Mitochondria were examined by transmission electron microscopy. Mitochondrial DNA copy number and deletions were assayed using long-range PCR. Mitochondrial membrane potential was measured by tetramethylrhodamine methyl ester uptake, and mitochondrial fragmentation was assessed by confocal microscopy. Exome sequencing was used to screen for pathogenic variants. RESULTS: OPA1 haploinsufficient iPSCs differentiated into dopaminergic neurons and exhibited marked reduction in OPA1 protein levels. Loss of OPA1 caused a late defect in oxidative phosphorylation, reduced complex I levels, and activity without a significant change in the ultrastructure of mitochondria. Loss of neurons in culture recapitulated dopaminergic degeneration in syndromic disease and correlated with mitochondrial fragmentation. INTERPRETATION: OPA1 levels maintain oxidative phosphorylation in iPSC-derived neurons, at least in part, by regulating the stability of complex I. Severity of OPA1 disease associates primarily with the extent of OPA1-mediated fusion, suggesting that activation of this mechanism or identification of its genetic modifiers may have therapeutic or prognostic value. Ann Neurol 2018;83:915-925.

Description

Keywords

DNA, Mitochondrial, GTP Phosphohydrolases, Humans, Induced Pluripotent Stem Cells, Membrane Potential, Mitochondrial, Mitochondria, Optic Atrophy, Oxidative Phosphorylation, Parkinsonian Disorders

Journal Title

Ann Neurol

Conference Name

Journal ISSN

0364-5134
1531-8249

Volume Title

83

Publisher

Wiley
Sponsorship
Department of Health (via National Institute for Health Research (NIHR)) (unknown)
Wellcome Trust (101876/Z/13/Z)
Wellcome Trust (101876/B/13/A)
Department of Health (via Newcastle University) (BH091682-RG81903)
Medical Research Council (MR/M024962/1)
Medical Research Council (MR/L023784/1)