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Novel engineered nanobodies specific for N‐terminal region of alpha‐synuclein recognize Lewy‐body pathology and inhibit in‐vitro seeded aggregation and toxicity

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submitted on 2024-06-26, 08:26 and posted on 2024-06-26, 08:28 authored by Issam Hmila, Nishant N. Vaikath, Nour K. Majbour, Daniel Erskine, Indulekha P. Sudhakaran, Vijay Gupta, Simona S. Ghanem, Zeyaul Islam, Mohamed M. Emara, Houari B. Abdesselem, Prasanna R. Kolatkar, Devaya K. Achappa, Tatiana Vinardell, Omar M. A. El‐Agnaf

Nanobodies (Nbs), the single‐domain antigen‐binding fragments of dromedary heavy‐chain antibodies (HCAb), are excellent candidates as therapeutic and diagnostic tools in synucleinopathies because of their small size, solubility and stability. Here, we constructed an immune nanobody library specific to the monomeric form of alpha‐synuclein (α‐syn). Phage display screening of the library allowed the identification of a nanobody, Nbα‐syn01, specific for α‐syn. Unlike previously developed nanobodies, Nbα‐syn01 recognized the N‐terminal region which is critical for in vitro and in vivo aggregation and contains many point mutations involved in early PD cases. The affinity of the monovalent Nbα‐syn01 and the engineered bivalent format BivNbα‐syn01 measured by isothermal titration calorimetry revealed unexpected results where Nbα‐syn01 and its bivalent format recognized preferentially α‐syn fibrils compared to the monomeric form. Nbα‐syn01 and BivNbα‐syn01 were also able to inhibit α‐syn‐seeded aggregation in vitro and reduced α‐syn‐seeded aggregation and toxicity in cells showing their potential to reduce α‐syn pathology. Moreover, both nanobody formats were able to recognize Lewy‐body pathology in human post‐mortem brain tissue from PD and DLB cases. Additionally, we present evidence through structural docking that Nbα‐syn01 binds the N‐terminal region of the α‐syn aggregated form. Overall, these results highlight the potential of Nbα‐syn01 and BivNbα‐syn01 in developing into a diagnostic or a therapeutic tool for PD and related disorders.

Other Information

Published in: The FEBS Journal
License: http://creativecommons.org/licenses/by-nc-nd/4.0/
See article on publisher's website: https://dx.doi.org/10.1111/febs.16376

Additional institutions affiliated with: Equine Veterinary Medical Center - Al Shaqab

Funding

Hamad Bin Khalifa University, Qatar Biomedical Research Institute.

History

Language

  • English

Publisher

Wiley

Publication Year

  • 2022

License statement

This Item is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.

Institution affiliated with

  • Hamad Bin Khalifa University
  • College of Health and Life Sciences - HBKU
  • Qatar Biomedical Research Institute - HBKU
  • Neurological Disorders Research Center - QBRI
  • Diabetes Research Center - QBRI
  • Qatar University
  • Qatar University Health - QU
  • College of Medicine - QU HEALTH
  • Biomedical and Pharmaceutical Research Unit - QU HEALTH
  • Al Shaqab

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