USP14 inhibition enhances Parkin-independent mitophagy in iNeurons

Loss of proteostasis is well documented during physiological aging and depends on the progressive decline in the activity of two major degradative mechanisms: the ubiquitin-proteasome system (UPS) and the autophagy-lysosomal pathway. This decline in proteostasis is exacerbated in age-associated neur...

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Main Authors: Greta Bernardo, Miguel A. Prado, Anna Roshani Dashtmian, Mariavittoria Favaro, Sofia Mauri, Alice Borsetto, Elena Marchesan, Joao A. Paulo, Steve P. Gygi, Daniel J. Finley, Elena Ziviani
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Language:English
Published: Elsevier 2024-12-01
Series:Pharmacological Research
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Online Access:http://www.sciencedirect.com/science/article/pii/S1043661824004298
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author Greta Bernardo
Miguel A. Prado
Anna Roshani Dashtmian
Mariavittoria Favaro
Sofia Mauri
Alice Borsetto
Elena Marchesan
Joao A. Paulo
Steve P. Gygi
Daniel J. Finley
Elena Ziviani
author_facet Greta Bernardo
Miguel A. Prado
Anna Roshani Dashtmian
Mariavittoria Favaro
Sofia Mauri
Alice Borsetto
Elena Marchesan
Joao A. Paulo
Steve P. Gygi
Daniel J. Finley
Elena Ziviani
author_sort Greta Bernardo
collection DOAJ
description Loss of proteostasis is well documented during physiological aging and depends on the progressive decline in the activity of two major degradative mechanisms: the ubiquitin-proteasome system (UPS) and the autophagy-lysosomal pathway. This decline in proteostasis is exacerbated in age-associated neurodegenerative diseases, such as Parkinson’s Disease (PD). In PD, patients develop an accumulation of aggregated proteins and dysfunctional mitochondria, which leads to ROS production, neuroinflammation and neurodegeneration. We recently reported that inhibition of the deubiquitinating enzyme USP14, which is known to enhance both the UPS and autophagy, increases lifespan and rescues the pathological phenotype of two Drosophila models of PD. Studies on the effects of USP14 inhibition in mammalian neurons have not yet been conducted. To close this gap, we exploited iNeurons differentiated from human embryonic stem cells (hESCs), and investigated the effect of inhibiting USP14 in these cultured neurons. Quantitative global proteomics analysis performed following genetic ablation or pharmacological inhibition of USP14 demonstrated that USP14 loss of function specifically promotes mitochondrial autophagy in iNeurons. Biochemical and imaging data also showed that USP14 inhibition enhances mitophagy. The mitophagic effect of USP14 inhibition proved to be PINK1/Parkin- independent, instead relying on expression of the mitochondrial E3 Ubiquitin Ligase MITOL/MARCH5. Notably, USP14 inhibition normalized the mitochondrial defects of Parkin KO human neurons.
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spelling doaj-art-0e236267259b4ce9b24ea8286377cdf12025-08-20T01:57:51ZengElsevierPharmacological Research1096-11862024-12-0121010748410.1016/j.phrs.2024.107484USP14 inhibition enhances Parkin-independent mitophagy in iNeuronsGreta Bernardo0Miguel A. Prado1Anna Roshani Dashtmian2Mariavittoria Favaro3Sofia Mauri4Alice Borsetto5Elena Marchesan6Joao A. Paulo7Steve P. Gygi8Daniel J. Finley9Elena Ziviani10Department of Biology, University of Padova, Padova, ItalyDepartment of Cell Biology, Harvard Medical School, Boston, USA; Instituto de Investigación Sanitaria del Principado de Asturias (ISPA), Oviedo, SpainDepartment of Biology, University of Padova, Padova, ItalyDepartment of Biology, University of Padova, Padova, ItalyDepartment of Biology, University of Padova, Padova, ItalyDepartment of Biology, University of Padova, Padova, ItalyDepartment of Biology, University of Padova, Padova, ItalyDepartment of Cell Biology, Harvard Medical School, Boston, USADepartment of Cell Biology, Harvard Medical School, Boston, USADepartment of Cell Biology, Harvard Medical School, Boston, USADepartment of Biology, University of Padova, Padova, Italy; Corresponding author.Loss of proteostasis is well documented during physiological aging and depends on the progressive decline in the activity of two major degradative mechanisms: the ubiquitin-proteasome system (UPS) and the autophagy-lysosomal pathway. This decline in proteostasis is exacerbated in age-associated neurodegenerative diseases, such as Parkinson’s Disease (PD). In PD, patients develop an accumulation of aggregated proteins and dysfunctional mitochondria, which leads to ROS production, neuroinflammation and neurodegeneration. We recently reported that inhibition of the deubiquitinating enzyme USP14, which is known to enhance both the UPS and autophagy, increases lifespan and rescues the pathological phenotype of two Drosophila models of PD. Studies on the effects of USP14 inhibition in mammalian neurons have not yet been conducted. To close this gap, we exploited iNeurons differentiated from human embryonic stem cells (hESCs), and investigated the effect of inhibiting USP14 in these cultured neurons. Quantitative global proteomics analysis performed following genetic ablation or pharmacological inhibition of USP14 demonstrated that USP14 loss of function specifically promotes mitochondrial autophagy in iNeurons. Biochemical and imaging data also showed that USP14 inhibition enhances mitophagy. The mitophagic effect of USP14 inhibition proved to be PINK1/Parkin- independent, instead relying on expression of the mitochondrial E3 Ubiquitin Ligase MITOL/MARCH5. Notably, USP14 inhibition normalized the mitochondrial defects of Parkin KO human neurons.http://www.sciencedirect.com/science/article/pii/S1043661824004298USP14UPSAutophagyMitophagyPINK1Parkin
spellingShingle Greta Bernardo
Miguel A. Prado
Anna Roshani Dashtmian
Mariavittoria Favaro
Sofia Mauri
Alice Borsetto
Elena Marchesan
Joao A. Paulo
Steve P. Gygi
Daniel J. Finley
Elena Ziviani
USP14 inhibition enhances Parkin-independent mitophagy in iNeurons
Pharmacological Research
USP14
UPS
Autophagy
Mitophagy
PINK1
Parkin
title USP14 inhibition enhances Parkin-independent mitophagy in iNeurons
title_full USP14 inhibition enhances Parkin-independent mitophagy in iNeurons
title_fullStr USP14 inhibition enhances Parkin-independent mitophagy in iNeurons
title_full_unstemmed USP14 inhibition enhances Parkin-independent mitophagy in iNeurons
title_short USP14 inhibition enhances Parkin-independent mitophagy in iNeurons
title_sort usp14 inhibition enhances parkin independent mitophagy in ineurons
topic USP14
UPS
Autophagy
Mitophagy
PINK1
Parkin
url http://www.sciencedirect.com/science/article/pii/S1043661824004298
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