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Amyloid fibril proteomics of AD brains reveals modifiers of aggregation and toxicity

BACKGROUND: The accumulation of amyloid beta (Aβ) peptides in fibrils is prerequisite for Alzheimer’s disease (AD). Our understanding of the proteins that promote Aβ fibril formation and mediate neurotoxicity has been limited due to technical challenges in isolating pure amyloid fibrils from brain e...

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Autores principales: Upadhyay, Arun, Chhangani, Deepak, Rao, Nalini R., Kofler, Julia, Vassar, Robert, Rincon-Limas, Diego E., Savas, Jeffrey N.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10503190/
https://www.ncbi.nlm.nih.gov/pubmed/37710351
http://dx.doi.org/10.1186/s13024-023-00654-z
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author Upadhyay, Arun
Chhangani, Deepak
Rao, Nalini R.
Kofler, Julia
Vassar, Robert
Rincon-Limas, Diego E.
Savas, Jeffrey N.
author_facet Upadhyay, Arun
Chhangani, Deepak
Rao, Nalini R.
Kofler, Julia
Vassar, Robert
Rincon-Limas, Diego E.
Savas, Jeffrey N.
author_sort Upadhyay, Arun
collection PubMed
description BACKGROUND: The accumulation of amyloid beta (Aβ) peptides in fibrils is prerequisite for Alzheimer’s disease (AD). Our understanding of the proteins that promote Aβ fibril formation and mediate neurotoxicity has been limited due to technical challenges in isolating pure amyloid fibrils from brain extracts. METHODS: To investigate how amyloid fibrils form and cause neurotoxicity in AD brain, we developed a robust biochemical strategy. We benchmarked the success of our purifications using electron microscopy, amyloid dyes, and a large panel of Aβ immunoassays. Tandem mass-spectrometry based proteomic analysis workflows provided quantitative measures of the amyloid fibril proteome. These methods allowed us to compare amyloid fibril composition from human AD brains, three amyloid mouse models, transgenic Aβ42 flies, and Aβ42 seeded cultured neurons. RESULTS: Amyloid fibrils are primarily composed by Aβ42 and unexpectedly harbor Aβ38 but generally lack Aβ40 peptides. Multidimensional quantitative proteomics allowed us to redefine the fibril proteome by identifying 20 new amyloid-associated proteins. Notably, we confirmed 57 previously reported plaque-associated proteins. We validated a panel of these proteins as bona fide amyloid-interacting proteins using antibodies and orthogonal proteomic analysis. One metal-binding chaperone metallothionein-3 is tightly associated with amyloid fibrils and modulates fibril formation in vitro. Lastly, we used a transgenic Aβ42 fly model to test if knock down or over-expression of fibril-interacting gene homologues modifies neurotoxicity. Here, we could functionally validate 20 genes as modifiers of Aβ42 toxicity in vivo. CONCLUSIONS: These discoveries and subsequent confirmation indicate that fibril-associated proteins play a key role in amyloid formation and AD pathology. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s13024-023-00654-z.
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spelling pubmed-105031902023-09-16 Amyloid fibril proteomics of AD brains reveals modifiers of aggregation and toxicity Upadhyay, Arun Chhangani, Deepak Rao, Nalini R. Kofler, Julia Vassar, Robert Rincon-Limas, Diego E. Savas, Jeffrey N. Mol Neurodegener Research Article BACKGROUND: The accumulation of amyloid beta (Aβ) peptides in fibrils is prerequisite for Alzheimer’s disease (AD). Our understanding of the proteins that promote Aβ fibril formation and mediate neurotoxicity has been limited due to technical challenges in isolating pure amyloid fibrils from brain extracts. METHODS: To investigate how amyloid fibrils form and cause neurotoxicity in AD brain, we developed a robust biochemical strategy. We benchmarked the success of our purifications using electron microscopy, amyloid dyes, and a large panel of Aβ immunoassays. Tandem mass-spectrometry based proteomic analysis workflows provided quantitative measures of the amyloid fibril proteome. These methods allowed us to compare amyloid fibril composition from human AD brains, three amyloid mouse models, transgenic Aβ42 flies, and Aβ42 seeded cultured neurons. RESULTS: Amyloid fibrils are primarily composed by Aβ42 and unexpectedly harbor Aβ38 but generally lack Aβ40 peptides. Multidimensional quantitative proteomics allowed us to redefine the fibril proteome by identifying 20 new amyloid-associated proteins. Notably, we confirmed 57 previously reported plaque-associated proteins. We validated a panel of these proteins as bona fide amyloid-interacting proteins using antibodies and orthogonal proteomic analysis. One metal-binding chaperone metallothionein-3 is tightly associated with amyloid fibrils and modulates fibril formation in vitro. Lastly, we used a transgenic Aβ42 fly model to test if knock down or over-expression of fibril-interacting gene homologues modifies neurotoxicity. Here, we could functionally validate 20 genes as modifiers of Aβ42 toxicity in vivo. CONCLUSIONS: These discoveries and subsequent confirmation indicate that fibril-associated proteins play a key role in amyloid formation and AD pathology. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s13024-023-00654-z. BioMed Central 2023-09-14 /pmc/articles/PMC10503190/ /pubmed/37710351 http://dx.doi.org/10.1186/s13024-023-00654-z Text en © The Author(s) 2023 https://creativecommons.org/licenses/by/4.0/Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) . The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/ (https://creativecommons.org/publicdomain/zero/1.0/) ) applies to the data made available in this article, unless otherwise stated in a credit line to the data.
spellingShingle Research Article
Upadhyay, Arun
Chhangani, Deepak
Rao, Nalini R.
Kofler, Julia
Vassar, Robert
Rincon-Limas, Diego E.
Savas, Jeffrey N.
Amyloid fibril proteomics of AD brains reveals modifiers of aggregation and toxicity
title Amyloid fibril proteomics of AD brains reveals modifiers of aggregation and toxicity
title_full Amyloid fibril proteomics of AD brains reveals modifiers of aggregation and toxicity
title_fullStr Amyloid fibril proteomics of AD brains reveals modifiers of aggregation and toxicity
title_full_unstemmed Amyloid fibril proteomics of AD brains reveals modifiers of aggregation and toxicity
title_short Amyloid fibril proteomics of AD brains reveals modifiers of aggregation and toxicity
title_sort amyloid fibril proteomics of ad brains reveals modifiers of aggregation and toxicity
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10503190/
https://www.ncbi.nlm.nih.gov/pubmed/37710351
http://dx.doi.org/10.1186/s13024-023-00654-z
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