Selection and structural characterization of anti-TREM2 scFvs that reduce levels of shed ectodomain

Aleksandra Szykowska; Yu Chen; Thomas B Smith; Charlotta Preger; Jingjing Yang; Dongming Qian; Shubhashish M Mukhopadhyay; Edvard Wigren; Stephen J Neame; Susanne Gräslund; Helena Persson; Peter J Atkinson; Elena Di Daniel; Emma Mead; John Wang; John B Davis; Nicola A Burgess-Brown; Alex N Bullock

doi:10.1016/j.str.2021.06.010

Selection and structural characterization of anti-TREM2 scFvs that reduce levels of shed ectodomain

Structure. 2021 Nov 4;29(11):1241-1252.e5. doi: 10.1016/j.str.2021.06.010. Epub 2021 Jul 6.

Authors

Aleksandra Szykowska¹, Yu Chen², Thomas B Smith³, Charlotta Preger⁴, Jingjing Yang⁵, Dongming Qian⁵, Shubhashish M Mukhopadhyay¹, Edvard Wigren⁴, Stephen J Neame⁶, Susanne Gräslund⁴, Helena Persson⁷, Peter J Atkinson⁶, Elena Di Daniel³, Emma Mead³, John Wang², John B Davis⁸, Nicola A Burgess-Brown⁹, Alex N Bullock¹⁰

Affiliations

¹ Centre for Medicines Discovery, Nuffield Department of Medicine, University of Oxford, Old Road Campus Research Building, Roosevelt Drive, Oxford OX3 7DQ, UK.
² Eisai Inc., 35 Cambridgepark Drive, Cambridge, MA 02140, USA.
³ Alzheimer's Research UK Oxford Drug Discovery Institute, NDM Research Building, University of Oxford, Old Road Campus, Roosevelt Drive, Oxford OX3 7FZ, UK; Centre for Medicines Discovery, Nuffield Department of Medicine, University of Oxford, Old Road Campus Research Building, Roosevelt Drive, Oxford OX3 7DQ, UK.
⁴ Structural Genomics Consortium (SGC), Karolinska Institutet, Karolinska University Hospital, Division of Rheumatology, Department of Medicine Solna, 171 76 Stockholm, Sweden.
⁵ Viva Biotech Ltd., 334 Aidisheng Road, Zhangjiang High-Tech Park, Shanghai 201203, China.
⁶ Eisai Ltd., Mosquito Way, Hatfield AL10 9SN, UK.
⁷ Science for Life Laboratory, Drug Discovery and Development & School of Engineering Sciences in Chemistry, Biotechnology and Health, Royal Institute of Technology (KTH), Stockholm, Sweden.
⁸ Alzheimer's Research UK Oxford Drug Discovery Institute, NDM Research Building, University of Oxford, Old Road Campus, Roosevelt Drive, Oxford OX3 7FZ, UK; Centre for Medicines Discovery, Nuffield Department of Medicine, University of Oxford, Old Road Campus Research Building, Roosevelt Drive, Oxford OX3 7DQ, UK. Electronic address: john.davis@cmd.ox.ac.uk.
⁹ Centre for Medicines Discovery, Nuffield Department of Medicine, University of Oxford, Old Road Campus Research Building, Roosevelt Drive, Oxford OX3 7DQ, UK. Electronic address: nicola.burgess-brown@cmd.ox.ac.uk.
¹⁰ Centre for Medicines Discovery, Nuffield Department of Medicine, University of Oxford, Old Road Campus Research Building, Roosevelt Drive, Oxford OX3 7DQ, UK. Electronic address: alex.bullock@cmd.ox.ac.uk.

Abstract

Mutations in TREM2, a receptor expressed by microglia in the brain, are associated with an increased risk of neurodegeneration, including Alzheimer's disease. Numerous studies support a role for TREM2 in sensing damaging stimuli and triggering signaling cascades necessary for neuroprotection. Despite its significant role, ligands and regulators of TREM2 activation, and the mechanisms governing TREM2-dependent responses and its cleavage from the membrane, remain poorly characterized. Here, we present phage display generated antibody single-chain variable fragments (scFvs) to human TREM2 immunoglobulin-like domain. Co-crystal structures revealed the binding of two scFvs to an epitope on the TREM2 domain distal to the putative ligand-binding site. Enhanced functional activity was observed for oligomeric scFv species, which inhibited the production of soluble TREM2 in a HEK293 cell model. We hope that detailed characterization of their epitopes and properties will facilitate the use of these renewable binders as structural and functional biology tools for TREM2 research.

Keywords: Alzheimer's disease; TREM2; X-ray crystallography; microglia; single-chain variable fragments.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Epitopes / metabolism*
HEK293 Cells
Humans
Membrane Glycoproteins / metabolism*
Phagocytosis / physiology
Receptors, Immunologic / metabolism*
Single-Chain Antibodies

Substances

Epitopes
Membrane Glycoproteins
Receptors, Immunologic
Single-Chain Antibodies
TREM2 protein, human

Abstract

Publication types

MeSH terms

Substances

Grants and funding