Oxidative switch drives mitophagy defects in dopaminergic parkin mutant patient neurons

Abstract Mutations in PRKN are the most common cause of early onset Parkinson’s disease. Parkin is an E3 ubiquitin ligase, functioning in mitophagy. Mitochondrial abnormalities are present in PRKN mutant models. Patient derived neurons are a promising model in which to study pathogenic mechanisms an...

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Autores principales:	Aurelie Schwartzentruber, Camilla Boschian, Fernanda Martins Lopes, Monika A. Myszczynska, Elizabeth J. New, Julien Beyrath, Jan Smeitink, Laura Ferraiuolo, Heather Mortiboys
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2020
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/5d11f9239fc34740ace065b8c2911136
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spelling	oai:doaj.org-article:5d11f9239fc34740ace065b8c29111362021-12-02T18:48:00ZOxidative switch drives mitophagy defects in dopaminergic parkin mutant patient neurons10.1038/s41598-020-72345-42045-2322https://doaj.org/article/5d11f9239fc34740ace065b8c29111362020-09-01T00:00:00Zhttps://doi.org/10.1038/s41598-020-72345-4https://doaj.org/toc/2045-2322Abstract Mutations in PRKN are the most common cause of early onset Parkinson’s disease. Parkin is an E3 ubiquitin ligase, functioning in mitophagy. Mitochondrial abnormalities are present in PRKN mutant models. Patient derived neurons are a promising model in which to study pathogenic mechanisms and therapeutic targets. Here we generate induced neuronal progenitor cells from PRKN mutant patient fibroblasts with a high dopaminergic neuron yield. We reveal changing mitochondrial phenotypes as neurons undergo a metabolic switch during differentiation. Fibroblasts from 4 controls and 4 PRKN mutant patients were transformed into induced neuronal progenitor cells and subsequently differentiated into dopaminergic neurons. Mitochondrial morphology, function and mitophagy were evaluated using live cell fluorescent imaging, cellular ATP and reactive oxygen species production quantification. Direct conversion of control and PRKN mutant patient fibroblasts results in induced neuronal progenitor and their differentiation yields high percentage of dopaminergic neurons. We were able to observe changing mitochondrial phenotypes as neurons undergo a metabolic switch during differentiation. Our results show that when pre-neurons are glycolytic early in differentiation mitophagy is unimpaired by PRKN deficiency. However as neurons become oxidative phosphorylation dependent, mitophagy is severely impaired in the PRKN mutant patient neurons. These changes correlate with changes in mitochondrial function and morphology; resulting in lower neuron yield and altered neuronal morphology. Induced neuronal progenitor cell conversion can produce a high yield of dopaminergic neurons. The mitochondrial phenotype, including mitophagy status, is highly dependent on the metabolic status of the cell. Only when neurons are oxidative phosphorylation reliant the extent of mitochondrial abnormalities are identified. These data provide insight into cell specific effects of PRKN mutations, in particular in relation to mitophagy dependent disease phenotypes and provide avenues for alternative therapeutic approaches.Aurelie SchwartzentruberCamilla BoschianFernanda Martins LopesMonika A. MyszczynskaElizabeth J. NewJulien BeyrathJan SmeitinkLaura FerraiuoloHeather MortiboysNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 10, Iss 1, Pp 1-14 (2020)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Aurelie Schwartzentruber Camilla Boschian Fernanda Martins Lopes Monika A. Myszczynska Elizabeth J. New Julien Beyrath Jan Smeitink Laura Ferraiuolo Heather Mortiboys Oxidative switch drives mitophagy defects in dopaminergic parkin mutant patient neurons
description	Abstract Mutations in PRKN are the most common cause of early onset Parkinson’s disease. Parkin is an E3 ubiquitin ligase, functioning in mitophagy. Mitochondrial abnormalities are present in PRKN mutant models. Patient derived neurons are a promising model in which to study pathogenic mechanisms and therapeutic targets. Here we generate induced neuronal progenitor cells from PRKN mutant patient fibroblasts with a high dopaminergic neuron yield. We reveal changing mitochondrial phenotypes as neurons undergo a metabolic switch during differentiation. Fibroblasts from 4 controls and 4 PRKN mutant patients were transformed into induced neuronal progenitor cells and subsequently differentiated into dopaminergic neurons. Mitochondrial morphology, function and mitophagy were evaluated using live cell fluorescent imaging, cellular ATP and reactive oxygen species production quantification. Direct conversion of control and PRKN mutant patient fibroblasts results in induced neuronal progenitor and their differentiation yields high percentage of dopaminergic neurons. We were able to observe changing mitochondrial phenotypes as neurons undergo a metabolic switch during differentiation. Our results show that when pre-neurons are glycolytic early in differentiation mitophagy is unimpaired by PRKN deficiency. However as neurons become oxidative phosphorylation dependent, mitophagy is severely impaired in the PRKN mutant patient neurons. These changes correlate with changes in mitochondrial function and morphology; resulting in lower neuron yield and altered neuronal morphology. Induced neuronal progenitor cell conversion can produce a high yield of dopaminergic neurons. The mitochondrial phenotype, including mitophagy status, is highly dependent on the metabolic status of the cell. Only when neurons are oxidative phosphorylation reliant the extent of mitochondrial abnormalities are identified. These data provide insight into cell specific effects of PRKN mutations, in particular in relation to mitophagy dependent disease phenotypes and provide avenues for alternative therapeutic approaches.
format	article
author	Aurelie Schwartzentruber Camilla Boschian Fernanda Martins Lopes Monika A. Myszczynska Elizabeth J. New Julien Beyrath Jan Smeitink Laura Ferraiuolo Heather Mortiboys
author_facet	Aurelie Schwartzentruber Camilla Boschian Fernanda Martins Lopes Monika A. Myszczynska Elizabeth J. New Julien Beyrath Jan Smeitink Laura Ferraiuolo Heather Mortiboys
author_sort	Aurelie Schwartzentruber
title	Oxidative switch drives mitophagy defects in dopaminergic parkin mutant patient neurons
title_short	Oxidative switch drives mitophagy defects in dopaminergic parkin mutant patient neurons
title_full	Oxidative switch drives mitophagy defects in dopaminergic parkin mutant patient neurons
title_fullStr	Oxidative switch drives mitophagy defects in dopaminergic parkin mutant patient neurons
title_full_unstemmed	Oxidative switch drives mitophagy defects in dopaminergic parkin mutant patient neurons
title_sort	oxidative switch drives mitophagy defects in dopaminergic parkin mutant patient neurons
publisher	Nature Portfolio
publishDate	2020
url	https://doaj.org/article/5d11f9239fc34740ace065b8c2911136
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Oxidative switch drives mitophagy defects in dopaminergic parkin mutant patient neurons

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