Quantitative Profiling of Nanoscopic Protein Aggregates Reveals Specific Fingerprint of TDP-43-Positive Assemblies in Motor Neuron Disease

Dezerae Cox; Melanie Burke; Sara Milani; Matthew A. White; Fergal M. Waldron; Dorothea Böken; Evgeniia Lobanova; Jemeen Sreedharan; Jenna M. Gregory; David Klenerman

doi:10.1002/advs.202505484

Quantitative Profiling of Nanoscopic Protein Aggregates Reveals Specific Fingerprint of TDP-43-Positive Assemblies in Motor Neuron Disease

Dezerae Cox^* (Corresponding Author)
, Melanie Burke
, Sara Milani
, Matthew A. White
, Fergal M. Waldron
, Dorothea Böken
, Evgeniia Lobanova
, Jemeen Sreedharan
, Jenna M. Gregory
, David Klenerman^* (Corresponding Author)

^*Corresponding author for this work

Research output: Contribution to journal › Article › peer-review

Abstract

Abnormal aggregation of TAR DNA-binding protein 43 (TDP-43) is a pathological hallmark of motor neuron disease (MND), yet current methods for quantifying these aggregates in biological samples remain limited in sensitivity and resolution. Here, single-molecule fluorescence microscopy is applied to post-mortem brain extracts to quantitatively characterize aggregates containing TDP-43 at the individual particle level. The resulting aggregate fingerprints, consisting of morphological and compositional profiles, are sufficient to distinguish MND donors from neurologically normal controls and further discriminate between clinically distinct MND subgroups. Comparative proteomic analysis confirms and extends these findings, revealing convergent and complementary molecular signatures. These results demonstrate, for the first time, that single-molecule aggregate profiling can stratify MND cases using patient-derived tissues, paving the way for the development of sensitive minimally invasive diagnostics and mechanistically informed disease monitoring tools.

Original language	English
Article number	e05484
Number of pages	13
Journal	Advanced Science
Volume	12
Issue number	44
Early online date	23 Sept 2025
DOIs	https://doi.org/10.1002/advs.202505484
Publication status	Published - 27 Nov 2025

Bibliographical note

The authors were grateful to all donors, their family members and caregivers, for making this study possible. We acknowledge the MRC London Neurodegenerative Diseases Brain Bank and the Brains for Dementia Research project specifically Dr Claire Troakes, for facilitating the collection, storage and provisioning of donated tissues.

Data Availability Statement

The data that support the findings of this study are openly available in PRIDE at https://www.ebi.ac.uk/pride/, reference number 61429.

Funding

This work was supported by the UK Dementia Research Institute, which receives its funding from DRI Ltd., funded by the UK Medical Research Council, Alzheimer's Society, and Alzheimer's Research. We would also like to acknowledge the support of the UK Dementia Research Institute Proteomics Platform, led by Dr Bethany Geary, and technical support in the form of reagents provided by Dr Yu P. Zhang (University of Cambridge). D.C. was supported by the Lady Edith Wolfson Junior Non-Clinical Research Fellowship awarded by the MND Association UK (Cox 971-799). JS was supported by a Wellcome Trust Senior Research Fellowship (221824/Z/20/Z). JMG was supported by a Motor Neurone Disease Association grant (Gregory/APR24/2374-791), and both JMG and FMW were supported by a Target ALS grant (BB-2022-C4-L2). MAW was supported by an Alzheimer's Research UK Research Fellowship (ARUK-RF2020A-008), an award from The Sean M. Healey & AMG Center for ALS at Mass General and ALS FindingACure, and the Rosetrees Trust (CF2/100004). EL was supported by a grant from Parkinson's UK (G-1901).

Funders	Funder number
Alzheimer's Research UK	ARUK‐RF2020A‐008
Wellcome Trust	221824/Z/20/Z
Motor Neurone Disease Association	Gregory/APR24/2374‐791, BB‐2022‐C4‐L2, 971‐799
CF2/100004
Parkinson's UK	G‐1901

Keywords

motor neuron disease
protein aggregates
proteomics
single-molecule microscopy

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Cite this

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title = "Quantitative Profiling of Nanoscopic Protein Aggregates Reveals Specific Fingerprint of TDP-43-Positive Assemblies in Motor Neuron Disease",

abstract = "Abnormal aggregation of TAR DNA-binding protein 43 (TDP-43) is a pathological hallmark of motor neuron disease (MND), yet current methods for quantifying these aggregates in biological samples remain limited in sensitivity and resolution. Here, single-molecule fluorescence microscopy is applied to post-mortem brain extracts to quantitatively characterize aggregates containing TDP-43 at the individual particle level. The resulting aggregate fingerprints, consisting of morphological and compositional profiles, are sufficient to distinguish MND donors from neurologically normal controls and further discriminate between clinically distinct MND subgroups. Comparative proteomic analysis confirms and extends these findings, revealing convergent and complementary molecular signatures. These results demonstrate, for the first time, that single-molecule aggregate profiling can stratify MND cases using patient-derived tissues, paving the way for the development of sensitive minimally invasive diagnostics and mechanistically informed disease monitoring tools.",

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AU - Burke, Melanie

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AU - White, Matthew A.

AU - Waldron, Fergal M.

AU - Böken, Dorothea

AU - Lobanova, Evgeniia

AU - Sreedharan, Jemeen

AU - Gregory, Jenna M.

AU - Klenerman, David

N1 - The authors were grateful to all donors, their family members and caregivers, for making this study possible. We acknowledge the MRC London Neurodegenerative Diseases Brain Bank and the Brains for Dementia Research project specifically Dr Claire Troakes, for facilitating the collection, storage and provisioning of donated tissues.

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Quantitative Profiling of Nanoscopic Protein Aggregates Reveals Specific Fingerprint of TDP-43-Positive Assemblies in Motor Neuron Disease

Abstract

Bibliographical note

Data Availability Statement

Funding

Keywords

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