Lentiviral Vectors Expressing Chimeric NEDD4 Ubiquitin Ligases: An Innovative Approach for Interfering with Alpha-Synuclein Accumulation

One of the main pathological features of Parkinson’s disease (PD) is a diffuse accumulation of alpha-synuclein (aS) aggregates in neurons. The NEDD4 E3 Ub ligase promotes aS degradation by the endosomal–lysosomal route. Interestingly, NEDD4, as well as being a small molecule able to trigger its func...

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Autores principales:	Stefania Vogiatzis, Michele Celestino, Marta Trevisan, Gloria Magro, Claudia Del Vecchio, Deran Erdengiz, Giorgio Palù, Cristina Parolin, Kathleen Maguire-Zeiss, Arianna Calistri
Formato:	article
Lenguaje:	EN
Publicado:	MDPI AG 2021
Materias:	NEDD4 ubiquibodies alpha-synuclein Parkinson’s disease Biology (General) QH301-705.5
Acceso en línea:	https://doaj.org/article/09ea083f25574f9dbd8207066102ba19
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spelling	oai:doaj.org-article:09ea083f25574f9dbd8207066102ba192021-11-25T17:13:21ZLentiviral Vectors Expressing Chimeric NEDD4 Ubiquitin Ligases: An Innovative Approach for Interfering with Alpha-Synuclein Accumulation10.3390/cells101132562073-4409https://doaj.org/article/09ea083f25574f9dbd8207066102ba192021-11-01T00:00:00Zhttps://www.mdpi.com/2073-4409/10/11/3256https://doaj.org/toc/2073-4409One of the main pathological features of Parkinson’s disease (PD) is a diffuse accumulation of alpha-synuclein (aS) aggregates in neurons. The NEDD4 E3 Ub ligase promotes aS degradation by the endosomal–lysosomal route. Interestingly, NEDD4, as well as being a small molecule able to trigger its functions, is protective against human aS toxicity in evolutionary distant models. While pharmacological activation of E3 enzymes is not easy to achieve, their flexibility and the lack of “<i>consensus</i>” motifs for Ub-conjugation allow the development of engineered Ub-ligases, able to target proteins of interest. We developed lentiviral vectors, encoding well-characterized anti-human aS scFvs fused in frame to the NEDD4 catalytic domain (ubiquibodies), in order to target ubiquitinate aS. We demonstrate that, while all generated ubiquibodies bind to and ubiquitinate aS, the one directed against the non-amyloid component (NAC) of aS (Nac32HECT) affects aS’s intracellular levels. Furthermore, Nac32HECT expression partially rescues aS’s overexpression or mutation toxicity in neural stem cells. Overall, our data suggest that ubiquibodies, and Nac32HECT in particular, represent a valid platform for interfering with the effects of aS’s accumulation and aggregation in neurons.Stefania VogiatzisMichele CelestinoMarta TrevisanGloria MagroClaudia Del VecchioDeran ErdengizGiorgio PalùCristina ParolinKathleen Maguire-ZeissArianna CalistriMDPI AGarticleNEDD4ubiquibodiesalpha-synucleinParkinson’s diseaseBiology (General)QH301-705.5ENCells, Vol 10, Iss 3256, p 3256 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	NEDD4 ubiquibodies alpha-synuclein Parkinson’s disease Biology (General) QH301-705.5
spellingShingle	NEDD4 ubiquibodies alpha-synuclein Parkinson’s disease Biology (General) QH301-705.5 Stefania Vogiatzis Michele Celestino Marta Trevisan Gloria Magro Claudia Del Vecchio Deran Erdengiz Giorgio Palù Cristina Parolin Kathleen Maguire-Zeiss Arianna Calistri Lentiviral Vectors Expressing Chimeric NEDD4 Ubiquitin Ligases: An Innovative Approach for Interfering with Alpha-Synuclein Accumulation
description	One of the main pathological features of Parkinson’s disease (PD) is a diffuse accumulation of alpha-synuclein (aS) aggregates in neurons. The NEDD4 E3 Ub ligase promotes aS degradation by the endosomal–lysosomal route. Interestingly, NEDD4, as well as being a small molecule able to trigger its functions, is protective against human aS toxicity in evolutionary distant models. While pharmacological activation of E3 enzymes is not easy to achieve, their flexibility and the lack of “<i>consensus</i>” motifs for Ub-conjugation allow the development of engineered Ub-ligases, able to target proteins of interest. We developed lentiviral vectors, encoding well-characterized anti-human aS scFvs fused in frame to the NEDD4 catalytic domain (ubiquibodies), in order to target ubiquitinate aS. We demonstrate that, while all generated ubiquibodies bind to and ubiquitinate aS, the one directed against the non-amyloid component (NAC) of aS (Nac32HECT) affects aS’s intracellular levels. Furthermore, Nac32HECT expression partially rescues aS’s overexpression or mutation toxicity in neural stem cells. Overall, our data suggest that ubiquibodies, and Nac32HECT in particular, represent a valid platform for interfering with the effects of aS’s accumulation and aggregation in neurons.
format	article
author	Stefania Vogiatzis Michele Celestino Marta Trevisan Gloria Magro Claudia Del Vecchio Deran Erdengiz Giorgio Palù Cristina Parolin Kathleen Maguire-Zeiss Arianna Calistri
author_facet	Stefania Vogiatzis Michele Celestino Marta Trevisan Gloria Magro Claudia Del Vecchio Deran Erdengiz Giorgio Palù Cristina Parolin Kathleen Maguire-Zeiss Arianna Calistri
author_sort	Stefania Vogiatzis
title	Lentiviral Vectors Expressing Chimeric NEDD4 Ubiquitin Ligases: An Innovative Approach for Interfering with Alpha-Synuclein Accumulation
title_short	Lentiviral Vectors Expressing Chimeric NEDD4 Ubiquitin Ligases: An Innovative Approach for Interfering with Alpha-Synuclein Accumulation
title_full	Lentiviral Vectors Expressing Chimeric NEDD4 Ubiquitin Ligases: An Innovative Approach for Interfering with Alpha-Synuclein Accumulation
title_fullStr	Lentiviral Vectors Expressing Chimeric NEDD4 Ubiquitin Ligases: An Innovative Approach for Interfering with Alpha-Synuclein Accumulation
title_full_unstemmed	Lentiviral Vectors Expressing Chimeric NEDD4 Ubiquitin Ligases: An Innovative Approach for Interfering with Alpha-Synuclein Accumulation
title_sort	lentiviral vectors expressing chimeric nedd4 ubiquitin ligases: an innovative approach for interfering with alpha-synuclein accumulation
publisher	MDPI AG
publishDate	2021
url	https://doaj.org/article/09ea083f25574f9dbd8207066102ba19
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Lentiviral Vectors Expressing Chimeric NEDD4 Ubiquitin Ligases: An Innovative Approach for Interfering with Alpha-Synuclein Accumulation

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