Cargando…

Assessment of Aggregated and Exosome-Associated α-Synuclein in Brain Tissue and Cerebrospinal Fluid Using Specific Immunoassays

Even though it is currently well-established that α-synuclein aggregation is closely associated with the pathological events in Parkinson’s disease (PD) and several other neurodegenerative disorders, collectively called synucleinopathies, the mechanistic link between α-synuclein aggregates and the o...

Descripción completa

Detalles Bibliográficos
Autores principales: Anagnostou, Dimitrios, Sfakianaki, Garifalia, Melachroinou, Katerina, Soutos, Miltiadis, Constantinides, Vassilios, Vaikath, Nishant, Tsantzali, Ioanna, Paraskevas, George P., Agnaf, Omar El, Vekrellis, Kostas, Emmanouilidou, Evangelia
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10340737/
https://www.ncbi.nlm.nih.gov/pubmed/37443586
http://dx.doi.org/10.3390/diagnostics13132192
_version_ 1785072150981378048
author Anagnostou, Dimitrios
Sfakianaki, Garifalia
Melachroinou, Katerina
Soutos, Miltiadis
Constantinides, Vassilios
Vaikath, Nishant
Tsantzali, Ioanna
Paraskevas, George P.
Agnaf, Omar El
Vekrellis, Kostas
Emmanouilidou, Evangelia
author_facet Anagnostou, Dimitrios
Sfakianaki, Garifalia
Melachroinou, Katerina
Soutos, Miltiadis
Constantinides, Vassilios
Vaikath, Nishant
Tsantzali, Ioanna
Paraskevas, George P.
Agnaf, Omar El
Vekrellis, Kostas
Emmanouilidou, Evangelia
author_sort Anagnostou, Dimitrios
collection PubMed
description Even though it is currently well-established that α-synuclein aggregation is closely associated with the pathological events in Parkinson’s disease (PD) and several other neurodegenerative disorders, collectively called synucleinopathies, the mechanistic link between α-synuclein aggregates and the onset and progression of neurodegeneration in these diseases remain unclear. The process of aggregation initiates from a structurally distorted monomer that gradually oligomerizes to generate a repertoire of fibrillar and oligomeric multimers that deposit within diseased cells in the brain. Total α-synuclein has been proposed as a potential biomarker in PD, but most of the studies do not discriminate between distinct α-synuclein conformers. To correlate protein measurements to disease pathology, we have developed a conformation-specific ELISA method that selectively detects fibrillar and oligomeric forms of α-synuclein without cross-reacting with monomers. We have used this assay to determine the levels of aggregated α-synuclein in human and mouse brain tissue as well as in CSF and CSF-derived exosomes from patients with synucleinopathy and control subjects. Our results verify the ability of the new assay to detect aggregated α-synuclein in complex matrices and support the idea that the levels of these conformers are related to the age of onset in PD patients, while CSF analysis showed that these species exist in low abundance in CSF and CSF-derived exosomes. Future studies will be required to fully assess the diagnostic usefulness of this ELISA in synucleinopathies.
format Online
Article
Text
id pubmed-10340737
institution National Center for Biotechnology Information
language English
publishDate 2023
publisher MDPI
record_format MEDLINE/PubMed
spelling pubmed-103407372023-07-14 Assessment of Aggregated and Exosome-Associated α-Synuclein in Brain Tissue and Cerebrospinal Fluid Using Specific Immunoassays Anagnostou, Dimitrios Sfakianaki, Garifalia Melachroinou, Katerina Soutos, Miltiadis Constantinides, Vassilios Vaikath, Nishant Tsantzali, Ioanna Paraskevas, George P. Agnaf, Omar El Vekrellis, Kostas Emmanouilidou, Evangelia Diagnostics (Basel) Article Even though it is currently well-established that α-synuclein aggregation is closely associated with the pathological events in Parkinson’s disease (PD) and several other neurodegenerative disorders, collectively called synucleinopathies, the mechanistic link between α-synuclein aggregates and the onset and progression of neurodegeneration in these diseases remain unclear. The process of aggregation initiates from a structurally distorted monomer that gradually oligomerizes to generate a repertoire of fibrillar and oligomeric multimers that deposit within diseased cells in the brain. Total α-synuclein has been proposed as a potential biomarker in PD, but most of the studies do not discriminate between distinct α-synuclein conformers. To correlate protein measurements to disease pathology, we have developed a conformation-specific ELISA method that selectively detects fibrillar and oligomeric forms of α-synuclein without cross-reacting with monomers. We have used this assay to determine the levels of aggregated α-synuclein in human and mouse brain tissue as well as in CSF and CSF-derived exosomes from patients with synucleinopathy and control subjects. Our results verify the ability of the new assay to detect aggregated α-synuclein in complex matrices and support the idea that the levels of these conformers are related to the age of onset in PD patients, while CSF analysis showed that these species exist in low abundance in CSF and CSF-derived exosomes. Future studies will be required to fully assess the diagnostic usefulness of this ELISA in synucleinopathies. MDPI 2023-06-27 /pmc/articles/PMC10340737/ /pubmed/37443586 http://dx.doi.org/10.3390/diagnostics13132192 Text en © 2023 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Anagnostou, Dimitrios
Sfakianaki, Garifalia
Melachroinou, Katerina
Soutos, Miltiadis
Constantinides, Vassilios
Vaikath, Nishant
Tsantzali, Ioanna
Paraskevas, George P.
Agnaf, Omar El
Vekrellis, Kostas
Emmanouilidou, Evangelia
Assessment of Aggregated and Exosome-Associated α-Synuclein in Brain Tissue and Cerebrospinal Fluid Using Specific Immunoassays
title Assessment of Aggregated and Exosome-Associated α-Synuclein in Brain Tissue and Cerebrospinal Fluid Using Specific Immunoassays
title_full Assessment of Aggregated and Exosome-Associated α-Synuclein in Brain Tissue and Cerebrospinal Fluid Using Specific Immunoassays
title_fullStr Assessment of Aggregated and Exosome-Associated α-Synuclein in Brain Tissue and Cerebrospinal Fluid Using Specific Immunoassays
title_full_unstemmed Assessment of Aggregated and Exosome-Associated α-Synuclein in Brain Tissue and Cerebrospinal Fluid Using Specific Immunoassays
title_short Assessment of Aggregated and Exosome-Associated α-Synuclein in Brain Tissue and Cerebrospinal Fluid Using Specific Immunoassays
title_sort assessment of aggregated and exosome-associated α-synuclein in brain tissue and cerebrospinal fluid using specific immunoassays
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10340737/
https://www.ncbi.nlm.nih.gov/pubmed/37443586
http://dx.doi.org/10.3390/diagnostics13132192
work_keys_str_mv AT anagnostoudimitrios assessmentofaggregatedandexosomeassociatedasynucleininbraintissueandcerebrospinalfluidusingspecificimmunoassays
AT sfakianakigarifalia assessmentofaggregatedandexosomeassociatedasynucleininbraintissueandcerebrospinalfluidusingspecificimmunoassays
AT melachroinoukaterina assessmentofaggregatedandexosomeassociatedasynucleininbraintissueandcerebrospinalfluidusingspecificimmunoassays
AT soutosmiltiadis assessmentofaggregatedandexosomeassociatedasynucleininbraintissueandcerebrospinalfluidusingspecificimmunoassays
AT constantinidesvassilios assessmentofaggregatedandexosomeassociatedasynucleininbraintissueandcerebrospinalfluidusingspecificimmunoassays
AT vaikathnishant assessmentofaggregatedandexosomeassociatedasynucleininbraintissueandcerebrospinalfluidusingspecificimmunoassays
AT tsantzaliioanna assessmentofaggregatedandexosomeassociatedasynucleininbraintissueandcerebrospinalfluidusingspecificimmunoassays
AT paraskevasgeorgep assessmentofaggregatedandexosomeassociatedasynucleininbraintissueandcerebrospinalfluidusingspecificimmunoassays
AT agnafomarel assessmentofaggregatedandexosomeassociatedasynucleininbraintissueandcerebrospinalfluidusingspecificimmunoassays
AT vekrelliskostas assessmentofaggregatedandexosomeassociatedasynucleininbraintissueandcerebrospinalfluidusingspecificimmunoassays
AT emmanouilidouevangelia assessmentofaggregatedandexosomeassociatedasynucleininbraintissueandcerebrospinalfluidusingspecificimmunoassays