Neurotoxin-mediated ­­potent activation of the axon degeneration regulator SARM1

  1. Andrea Loreto  Is a corresponding author
  2. Carlo Angeletti
  3. Weixi Gu
  4. Andrew Osborne
  5. Bart Nieuwenhuis
  6. Jonathan Gilley
  7. Peter Arthur-Farraj
  8. Elisa Merlini
  9. Adolfo Amici
  10. Zhenyao Luo
  11. Lauren Hartley-Tassell
  12. Thomas Ve
  13. Laura M Desrochers
  14. Qi Wang
  15. Bostjan Kobe
  16. Giuseppe Orsomando  Is a corresponding author
  17. Michael P Coleman  Is a corresponding author
  1. University of Cambridge, United Kingdom
  2. Polytechnic University of Marche, Italy
  3. University of Queensland, Australia
  4. Griffith University, Australia
  5. AstraZeneca, United States
  6. The University of Queensland, Australia

Abstract

Axon loss underlies symptom onset and progression in many neurodegenerative disorders. Axon degeneration in injury and disease is promoted by activation of the nicotinamide adenine dinucleotide (NAD)-consuming enzyme SARM1. Here, we report a novel activator of SARM1, a metabolite of the pesticide and neurotoxin vacor. Removal of SARM1 completely rescues mouse neurons from vacor-induced neuron and axon death in vitro and in vivo. We present the crystal structure the Drosophila SARM1 regulatory domain complexed with this activator, the vacor metabolite VMN, which as the most potent activator yet know is likely to support drug development for human SARM1 and NMNAT2 disorders. This study indicates the mechanism of neurotoxicity and pesticide action by vacor, raises important questions about other pyridines in wider use today, provides important new tools for drug discovery, and demonstrates that removing SARM1 can robustly block programmed axon death induced by toxicity as well as genetic mutation.

Data availability

All data generated or analysed during this study are included in the manuscript and supporting file; Source Data files have been provided for Figures 1-7 and figure supplements. VMN-bound dSARM1ARM crystal structure has been deposited in the Protein Data Bank (PDB: 7M6K).

The following data sets were generated

Article and author information

Author details

  1. Andrea Loreto

    Department of Clinical Neurosciences, University of Cambridge, Cambridge, United Kingdom
    For correspondence
    al850@cam.ac.uk
    Competing interests
    No competing interests declared.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-6535-6436
  2. Carlo Angeletti

    Department of Clinical Sciences (DISCO), Polytechnic University of Marche, Ancona, Italy
    Competing interests
    No competing interests declared.
  3. Weixi Gu

    School of Chemistry and Molecular Biosciences, University of Queensland, Brisbane, Australia
    Competing interests
    Weixi Gu, receives research funding from Disarm Therapeutics, a wholly-owned subsidiary of Eli Lilly and Co., Cambridge, MA, USA, but they had no role in the research presented here..
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-1185-3557
  4. Andrew Osborne

    Department of Clinical Neurosciences, University of Cambridge, Cambridge, United Kingdom
    Competing interests
    Andrew Osborne, is affiliated with Ikarovec Ltd. The author has no financial interests to declare..
  5. Bart Nieuwenhuis

    Department of Clinical Neurosciences, University of Cambridge, Cambridge, United Kingdom
    Competing interests
    No competing interests declared.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-2065-2271
  6. Jonathan Gilley

    Department of Clinical Neurosciences, University of Cambridge, Cambridge, United Kingdom
    Competing interests
    No competing interests declared.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-9510-7956
  7. Peter Arthur-Farraj

    Department of Clinical Neurosciences, University of Cambridge, Cambridge, United Kingdom
    Competing interests
    No competing interests declared.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-1239-9392
  8. Elisa Merlini

    Department of Clinical Neurosciences, University of Cambridge, Cambridge, United Kingdom
    Competing interests
    No competing interests declared.
  9. Adolfo Amici

    Department of Clinical Sciences (DISCO), Polytechnic University of Marche, Ancona, Italy
    Competing interests
    No competing interests declared.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-1081-7749
  10. Zhenyao Luo

    School of Chemistry and Molecular Biosciences, University of Queensland, Brisbane, Australia
    Competing interests
    Zhenyao Luo, receives research funding from Disarm Therapeutics, a wholly- owned subsidiary of Eli Lilly and Co., Cambridge, MA, USA, but they had no role in the research presented here..
  11. Lauren Hartley-Tassell

    Institute for Glycomics, Griffith University, Southport, Australia
    Competing interests
    No competing interests declared.
  12. Thomas Ve

    Institute for Glycomics, Griffith University, Brisbane, Australia
    Competing interests
    Thomas Ve, Thomas Ve receives research funding from Disarm Therapeutics, a wholly-owned subsidiary of Eli Lilly & Co., Cambridge, MA, USA, but they had no role in the research presented here..
  13. Laura M Desrochers

    Neuroscience, BioPharmaceuticals R and D, AstraZeneca, Waltham, United States
    Competing interests
    Laura M Desrochers, This work is in part funded by a BBSRC/AstraZeneca Industrial Partnership Award and Laura M Desrochers was an employee of AstraZeneca for part of the project. Laura M Desrochers is affiliated with Vertex Pharmaceuticals. The author has no financial interests to declare..
  14. Qi Wang

    Neuroscience, BioPharmaceuticals R and D, AstraZeneca, Waltham, United States
    Competing interests
    Qi Wang, This work is in part funded by a BBSRC/AstraZeneca Industrial Partnership Award and Qi Wang was an employee of AstraZeneca for part of the project. Qi Wang is affiliated with Kymera Therapeutics. The author has no financial interests to declare..
  15. Bostjan Kobe

    School of Chemistry and Molecular Biosciences, The University of Queensland, Brisbane, Australia
    Competing interests
    Bostjan Kobe, receives research funding from Disarm Therapeutics, a wholly-owned subsidiary of Eli Lilly and Co., Cambridge, MA, USA, but they had no role in the research presented here..
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-9413-9166
  16. Giuseppe Orsomando

    Department of Clinical Sciences (DISCO), Polytechnic University of Marche, Ancona, Italy
    For correspondence
    g.orsomando@staff.univpm.it
    Competing interests
    No competing interests declared.
  17. Michael P Coleman

    Department of Clinical Neurosciences, University of Cambridge, Cambridge, United Kingdom
    For correspondence
    mc469@cam.ac.uk
    Competing interests
    Michael P Coleman, holds funding jointly provided by AstraZeneca for academic research and consults for Nura Bio, but they had no role in the research presented here..

Funding

Wellcome Trust (210904/Z/18/Z)

  • Andrea Loreto

Wellcome Trust (206634)

  • Peter Arthur-Farraj

Biotechnology and Biological Sciences Research Council (BB/S009582/1)

  • Andrea Loreto
  • Jonathan Gilley
  • Michael P Coleman

UNIVPM (RSA 2016-18 and 2017-19)

  • Giuseppe Orsomando

Australian National Health and Medical Research Council (NHMRC 1160570)

  • Bostjan Kobe

Sight Research UK (SAC 041)

  • Andrew Osborne
  • Bart Nieuwenhuis

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.

Ethics

Animal experimentation: All studies conformed to the institution's ethical requirements in accordance with the 1986 Animals (Scientific Procedures) Act under Project Licences PPL P98A03BF9 and PP1824519, and in accordance with the Association for Research in Vision and Ophthalmology's Statement for the Use of Animals in Ophthalmic and Visual Research.

Copyright

© 2021, Loreto et al.

This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.

Metrics

  • 4,364
    views
  • 749
    downloads
  • 40
    citations

Views, downloads and citations are aggregated across all versions of this paper published by eLife.

Download links

A two-part list of links to download the article, or parts of the article, in various formats.

Downloads (link to download the article as PDF)

Open citations (links to open the citations from this article in various online reference manager services)

Cite this article (links to download the citations from this article in formats compatible with various reference manager tools)

  1. Andrea Loreto
  2. Carlo Angeletti
  3. Weixi Gu
  4. Andrew Osborne
  5. Bart Nieuwenhuis
  6. Jonathan Gilley
  7. Peter Arthur-Farraj
  8. Elisa Merlini
  9. Adolfo Amici
  10. Zhenyao Luo
  11. Lauren Hartley-Tassell
  12. Thomas Ve
  13. Laura M Desrochers
  14. Qi Wang
  15. Bostjan Kobe
  16. Giuseppe Orsomando
  17. Michael P Coleman
(2021)
Neurotoxin-mediated ­­potent activation of the axon degeneration regulator SARM1
eLife 10:e72823.
https://doi.org/10.7554/eLife.72823

Share this article

https://doi.org/10.7554/eLife.72823

Further reading

    1. Biochemistry and Chemical Biology
    2. Microbiology and Infectious Disease
    Mai Nguyen, Elda Bauda ... Cecile Morlot
    Research Article

    Teichoic acids (TA) are linear phospho-saccharidic polymers and important constituents of the cell envelope of Gram-positive bacteria, either bound to the peptidoglycan as wall teichoic acids (WTA) or to the membrane as lipoteichoic acids (LTA). The composition of TA varies greatly but the presence of both WTA and LTA is highly conserved, hinting at an underlying fundamental function that is distinct from their specific roles in diverse organisms. We report the observation of a periplasmic space in Streptococcus pneumoniae by cryo-electron microscopy of vitreous sections. The thickness and appearance of this region change upon deletion of genes involved in the attachment of TA, supporting their role in the maintenance of a periplasmic space in Gram-positive bacteria as a possible universal function. Consequences of these mutations were further examined by super-resolved microscopy, following metabolic labeling and fluorophore coupling by click chemistry. This novel labeling method also enabled in-gel analysis of cell fractions. With this approach, we were able to titrate the actual amount of TA per cell and to determine the ratio of WTA to LTA. In addition, we followed the change of TA length during growth phases, and discovered that a mutant devoid of LTA accumulates the membrane-bound polymerized TA precursor.

    1. Biochemistry and Chemical Biology
    2. Computational and Systems Biology
    Shinichi Kawaguchi, Xin Xu ... Toshie Kai
    Research Article

    Protein–protein interactions are fundamental to understanding the molecular functions and regulation of proteins. Despite the availability of extensive databases, many interactions remain uncharacterized due to the labor-intensive nature of experimental validation. In this study, we utilized the AlphaFold2 program to predict interactions among proteins localized in the nuage, a germline-specific non-membrane organelle essential for piRNA biogenesis in Drosophila. We screened 20 nuage proteins for 1:1 interactions and predicted dimer structures. Among these, five represented novel interaction candidates. Three pairs, including Spn-E_Squ, were verified by co-immunoprecipitation. Disruption of the salt bridges at the Spn-E_Squ interface confirmed their functional importance, underscoring the predictive model’s accuracy. We extended our analysis to include interactions between three representative nuage components—Vas, Squ, and Tej—and approximately 430 oogenesis-related proteins. Co-immunoprecipitation verified interactions for three pairs: Mei-W68_Squ, CSN3_Squ, and Pka-C1_Tej. Furthermore, we screened the majority of Drosophila proteins (~12,000) for potential interaction with the Piwi protein, a central player in the piRNA pathway, identifying 164 pairs as potential binding partners. This in silico approach not only efficiently identifies potential interaction partners but also significantly bridges the gap by facilitating the integration of bioinformatics and experimental biology.