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Biased M1-muscarinic-receptor-mutant mice inform the design of next-generation drugs

Nature Chemical Biology volume 16, pages 240–249 (2020)Cite this article

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Abstract

Cholinesterase inhibitors, the current frontline symptomatic treatment for Alzheimer’s disease (AD), are associated with low efficacy and adverse effects. M1 muscarinic acetylcholine receptors (M1 mAChRs) represent a potential alternate therapeutic target; however, drug discovery programs focused on this G protein-coupled receptor (GPCR) have failed, largely due to cholinergic adverse responses. Employing novel chemogenetic and phosphorylation-deficient, G protein-biased, mouse models, paired with a toolbox of probe molecules, we establish previously unappreciated pharmacologically targetable M1 mAChR neurological processes, including anxiety-like behaviors and hyper-locomotion. By mapping the upstream signaling pathways regulating these responses, we determine the importance of receptor phosphorylation-dependent signaling in driving clinically relevant outcomes and in controlling adverse effects including ‘epileptic-like’ seizures. We conclude that M1 mAChR ligands that promote receptor phosphorylation-dependent signaling would protect against cholinergic adverse effects in addition to driving beneficial responses such as learning and memory and anxiolytic behavior relevant for the treatment of AD.

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Fig. 1: Generation of the M1-DREADD knock-in mouse.
Fig. 2: Altered phenotypes of M1-DREADD mice are corrected by CNO.
Fig. 3: Generation of the G protein-biased M1-PD knock-in mouse.
Fig. 4: Mapping of bimodal signaling to M1 mAChR physiological responses.
Fig. 5: M1-DREADD PD mice show epileptic-like seizures.
Fig. 6: Pilocarpine shows G protein bias, whereas GSK1034702 is not biased.

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Data availability

All data are available from the authors or are available through the University of Glasgow online data repository.

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Acknowledgements

This work is funded by a University of Glasgow Lord Kelvin Adam Smith Fellowship (to S.J.B.), an MRC MICA agreement MR/P019366/1 (to A.B.T., S.J.B., M.R. and C.M.), a Wellcome Trust Collaborative Award 201529/Z/16/Z (to A.B.T., P.V., A.C., L.D., C.M. and P.M.S.), SULSA Dementia Seed Funding (to A.B.T., P.V. and S.S.), ARUK PPG2017B-005 (to S.S.) and an MRC iCASE studentship with Eli Lilly (M.S.). This work was also supported by an MRC group leader program awarded to A.B.T. at the MRC Toxicology Unit. We acknowledge support from the BSU facilities at the Cancer Research UK Beatson Institute (C596/A17196) and the Biological Services at the University of Glasgow. We also thank M. Gaellman for support to the Tobin laboratory.

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Authors and Affiliations

  1. The Centre for Translational Pharmacology, Institute of Molecular, Cell and Systems Biology, College of Medical, Veterinary and Life Sciences, University of Glasgow, Glasgow, UK

    Sophie J. Bradley, Colin Molloy, Paulina Valuskova, Louis Dwomoh, Miriam Scarpa, Mario Rossi, Lisa Finlayson & Andrew B. Tobin

  2. Eli Lilly & Co, Neuroscience Discovery, Lilly Corporate Center, Indianapolis, IN, USA

    Kjell A. Svensson, Eyassu Chernet, Vanessa N. Barth & Adrian J. Mogg

  3. School of Life Sciences, Queen’s Medical Centre, University of Nottingham, Nottingham, UK

    Karolina Gherbi, David A. Sykes & Elizabeth M. Rosethorne

  4. Excellerate Bioscience Ltd, BioCity, Nottingham, UK

    Karolina Gherbi

  5. Centre of Membrane and Protein and Receptors (COMPARE), University of Birmingham and University of Nottingham, Midlands, UK

    David A. Sykes & Elizabeth M. Rosethorne

  6. Strathclyde Institute of Pharmacy and Biomedical Sciences, University of Strathclyde, Glasgow, UK

    Caroline A. Wilson & Shuzo Sakata

  7. Department of Molecular and Cell Biology, Henry Wellcome Building, University of Leicester, Leicester, UK

    Rajendra Mistry & R. A. John Challiss

  8. Drug Discovery Biology, Monash Institute of Pharmaceutical Sciences and Department of Pharmacology, Monash University, Parkville, Victoria, Australia

    Patrick M. Sexton & Arthur Christopoulos

  9. Eli Lilly & Co, Neuroscience Discovery, Erl Wood Manor, Windlesham, Surrey, UK

    Lisa M. Broad

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Contributions

S.J.B. and A.B.T. devised the program of work and wrote the paper with assistance from all other authors. S.J.B., A.B.T., R.A.J.C., L.M.B., E.M.R., S.S., P.M.S., A.J.M. and A.C. designed and advised on experiments. L.D. conducted receptor phosphorylation studies. C.M. and L.F. managed the mouse colony and conducted mouse behavioral experiments with S.J.B. P.V., C.A.W. and L.F. conducted the EEG recording studies. M.S., M.R. and R.M. conducted signaling and internalization studies. K.A.S., E.C. and V.N.B. conducted in vivo receptor occupancy and in vivo ligand activity assays. K.G. and D.A.S. conducted arrestin assays.

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Correspondence to Sophie J. Bradley or Andrew B. Tobin.

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Bradley, S.J., Molloy, C., Valuskova, P. et al. Biased M1-muscarinic-receptor-mutant mice inform the design of next-generation drugs. Nat Chem Biol 16, 240–249 (2020). https://doi.org/10.1038/s41589-019-0453-9

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