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Elamipretide (SS-31) treatment attenuates age-associated post-translational modifications of heart proteins

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Abstract

It has been demonstrated that elamipretide (SS-31) rescues age-related functional deficits in the heart but the full set of mechanisms behind this have yet to be determined. We investigated the hypothesis that elamipretide influences post-translational modifications to heart proteins. The S-glutathionylation and phosphorylation proteomes of mouse hearts were analyzed using shotgun proteomics to assess the effects of aging on these post-translational modifications and the ability of the mitochondria-targeted drug elamipretide to reverse age-related changes. Aging led to an increase in oxidation of protein thiols demonstrated by increased S-glutathionylation of cysteine residues on proteins from Old (24 months old at the start of the study) mouse hearts compared to Young (5–6 months old). This shift in the oxidation state of the proteome was almost completely reversed by 8 weeks of treatment with elamipretide. Many of the significant changes that occurred were in proteins involved in mitochondrial or cardiac function. We also found changes in the mouse heart phosphoproteome that were associated with age, some of which were partially restored with elamipretide treatment. Parallel reaction monitoring of a subset of phosphorylation sites revealed that the unmodified peptide reporting for Myot S231 increased with age, but not its phosphorylated form and that both phosphorylated and unphosphorylated forms of the peptide covering cMyBP-C S307 increased, but that elamipretide treatment did not affect these changes. These results suggest that changes to thiol redox state and phosphorylation status are two ways in which age may affect mouse heart function, which can be restored by treatment with elamipretide.

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Acknowledgements

The thiol redox proteomics experiments described herein were performed in the Environmental Molecular Sciences Laboratory, Pacific Northwest National Laboratory, a national scientific user facility sponsored by the Department of Energy under Contract DE-AC05-76RL0 1830. Elamipretide was provided by Stealth Therapeutics (Newton, MA) free of charge. Stealth Therapeutics did not play any role in the experimental design, data collection, or authorship of this research.

Funding

Funding for this research was provided by the UW Genetic Approaches to Aging Training Grant (T32AG000057-40), NIH/NIA grant P01AG001751, NIH/NIEHS grant P30ES007033, and the UW Nathan Shock Center.

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Author notes

  1. Jeremy A. Whitson

    Present address: Department of Biology, Davidson College, 405 N Main St, Davidson, NC, 28035, USA

Authors and Affiliations

  1. Department of Pathology, University of Washington, 1959 NE Pacific St, Seattle, WA, 98195, USA

    Jeremy A. Whitson & Peter S. Rabinovitch

  2. Department of Genome Sciences, University of Washington, 3720 15th Street NE, Seattle, WA, 98195, USA

    Miguel Martín-Pérez, Gennifer E. Merrihew, Eric Huang, Michael J. MacCoss & Judit Villén

  3. Integrative Omics, Pacific Northwest National Laboratory, 902 Battelle Boulevard, Richland, WA, 99352, USA

    Tong Zhang, Matthew J. Gaffrey & Wei-Jun Qian

  4. Department of Environmental & Occupational Health Sciences, University of Washington, 4225 Roosevelt Way NE, Seattle, WA, 98105, USA

    Collin C. White & Terrance J. Kavanagh

  5. Department of Radiology, University of Washington, 1959 NE Pacific St, Seattle, WA, 98195, USA

    Matthew D. Campbell & David J. Marcinek

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  1. Jeremy A. Whitson
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Correspondence to Peter S. Rabinovitch.

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Whitson, J., Martín-Pérez, M., Zhang, T. et al. Elamipretide (SS-31) treatment attenuates age-associated post-translational modifications of heart proteins. GeroScience 43, 2395–2412 (2021). https://doi.org/10.1007/s11357-021-00447-6

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