High-throughput discovery of fluoroprobes that recognize amyloid fibril polymorphs
Aggregation of microtubule-associated protein tau into conformationally distinct fibrils underpins neurodegenerative tauopathies. Fluorescent probes (fluoroprobes) such as thioflavin T have been essential tools for studying tau aggregation; however, most of them do not discriminate between amyloid fibril conformations (polymorphs). This gap is due, in part, to a lack of high-throughput methods for screening large, diverse chemical collections. Here we leverage advances in protein-adaptive differential scanning fluorimetry to screen the Aurora collection of 300+ fluoroprobes against multiple synthetic fibril polymorphs, including those formed from tau, α-synuclein and islet amyloid polypeptide. This screen-coupled with excitation-multiplexed bright-emission recording (EMBER) imaging and orthogonal secondary assays-revealed pan-fibril-binding chemotypes, as well as fluoroprobes selective for fibril subsets. One fluoroprobe recognized tau pathology in ex vivo brain slices from Alzheimer's disease and rodent models. We propose that these scaffolds represent entry points for developing fibril-selective ligands.
Authors
- Kelly M. Montgomery
- Hyunjun Yang (ORCID: https://orcid.org/0000-0003-2524-2201)
- William F. DeGrado (ORCID: https://orcid.org/0000-0003-4745-263X)
- Neil A. Ranson (ORCID: https://orcid.org/0000-0002-3640-5275)
- Martin Wilkinson (ORCID: https://orcid.org/0000-0001-5490-613X)
- Zoe Millbern (ORCID: https://orcid.org/0000-0002-0727-4261)
- Julia P. G. Jones (ORCID: https://orcid.org/0000-0002-5199-3335)
- Nelson R. Vinueza (ORCID: https://orcid.org/0000-0002-2608-5431)
- Jason E. Gestwicki (ORCID: https://orcid.org/0000-0002-6125-3154)
- Abby Oehler
- Emma C. Carroll (ORCID: https://orcid.org/0000-0002-6766-4245)
- Sheena E. Radford (ORCID: https://orcid.org/0000-0002-3079-8039)
- Wyatt C. Powell (ORCID: https://orcid.org/0000-0002-1281-4605)
- Daniel A. Mordes
- Alexander I. P. Taylor (ORCID: https://orcid.org/0000-0003-3131-3619)
- Carlo Condello (ORCID: https://orcid.org/0000-0002-8454-5360)
- Annemarie F. Charvat (ORCID: https://orcid.org/0009-0002-1164-4816)
- Anthony Yung
Institutions
- University of Leeds (GB)
- North Carolina State University (US)
- University of California, San Francisco (US)
- Institute for Neurodegenerative Disorders (US)
- Institute of Structural and Molecular Biology (GB)
- San Jose State University (US)
- Brandeis University (US)
Publication Details
- Journal
- Nature Chemistry
- Published
- 2025-08-14
- DOI
- https://doi.org/10.1038/s41557-025-01889-7
- Citations
- 3
- Primary Topic
- Alzheimer's disease research and treatments
- Type
- article
- Field-Weighted Citation Impact
- 3.49
Funders
- Royal Society
- National Institutes of Health
- University of California, San Francisco
- Medical Research Council