Myofibroblast senescence promotes arrhythmogenic remodeling in the aged infarcted rabbit heart

  1. Brett C Baggett
  2. Kevin R Murphy
  3. Elif Sengun
  4. Eric Mi
  5. Yueming Cao
  6. Nilufer N Turan
  7. Yichun Lu
  8. Lorraine Scofield
  9. Tae Yun Kim
  10. Anatoli Y Kabakov
  11. Peter Bronk
  12. Zhilin Qu
  13. Patrizia Camelliti
  14. Patrycja Dubielecka
  15. Dmitry Terentyev
  16. Federica del Monte
  17. Bum-Rak Choi
  18. John Sedivy
  19. Gideon Koren  Is a corresponding author
  1. Brown University, United States
  2. Rhode Island Hospital, United States
  3. University of California, Los Angeles, United States
  4. University of Surrey, United Kingdom
  5. Medical University of South Carolina, United States

Abstract

Progressive tissue remodeling after myocardial infarction (MI) promotes cardiac arrhythmias. This process is well studied in young animals, but little is known about pro-arrhythmic changes in aged animals. Senescent cells accumulate with age and accelerate age-associated diseases. Senescent cells interfere with cardiac function and outcome post-MI with age, but studies have not been performed in larger animals, and the mechanisms are unknown. Specifically, age-associated changes in timecourse of senescence and related changes in inflammation and fibrosis are not well understood. Additionally, the cellular and systemic role of senescence and its inflammatory milieu in influencing arrhythmogenesis with age is not clear, particularly in large animal models with cardiac electrophysiology more similar to humans than previously studied animal models. Here, we investigated the role of senescence in regulating inflammation, fibrosis, and arrhythmogenesis in young and aged infarcted rabbits. Aged rabbits exhibited increased peri-procedural mortality and arrhythmogenic electrophysiological remodeling at the infarct border zone (IBZ) compared to young rabbits. Studies of the aged infarct zone revealed persistent myofibroblast senescence and increased inflammatory signaling over a twelve-week timecourse. Senescent IBZ myofibroblasts in aged rabbits appear to be coupled to myocytes, and our computational modeling showed that senescent myofibroblast-cardiomyocyte coupling prolongs action potential duration (APD) and facilitates conduction block permissive of arrhythmias. Aged infarcted human ventricles show levels of senescence consistent with aged rabbits, and senescent myofibroblasts also couple to IBZ myocytes. Our findings suggest that therapeutic interventions targeting senescent cells may mitigate arrhythmias post-MI with age.

Data availability

All data generated or analyzed during this study are included in the provided Source Data file.

Article and author information

Author details

  1. Brett C Baggett

    Brown University, Providence, United States
    Competing interests
    The authors declare that no competing interests exist.
  2. Kevin R Murphy

    Brown University, Providence, United States
    Competing interests
    The authors declare that no competing interests exist.
  3. Elif Sengun

    Brown University, Providence, United States
    Competing interests
    The authors declare that no competing interests exist.
  4. Eric Mi

    Brown University, Providence, United States
    Competing interests
    The authors declare that no competing interests exist.
  5. Yueming Cao

    Brown University, Providence, United States
    Competing interests
    The authors declare that no competing interests exist.
  6. Nilufer N Turan

    Cardiovascular Research Center, Rhode Island Hospital, Providence, United States
    Competing interests
    The authors declare that no competing interests exist.
  7. Yichun Lu

    Cardiovascular Research Center, Rhode Island Hospital, Providence, United States
    Competing interests
    The authors declare that no competing interests exist.
  8. Lorraine Scofield

    Cardiovascular Research Center, Rhode Island Hospital, Providence, United States
    Competing interests
    The authors declare that no competing interests exist.
  9. Tae Yun Kim

    Cardiovascular Research Center, Rhode Island Hospital, Providence, United States
    Competing interests
    The authors declare that no competing interests exist.
  10. Anatoli Y Kabakov

    Brown University, Providence, United States
    Competing interests
    The authors declare that no competing interests exist.
  11. Peter Bronk

    Cardiovascular Research Center, Rhode Island Hospital, Providence, United States
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-9067-2016
  12. Zhilin Qu

    School of Medicine, University of California, Los Angeles, Los Angeles, United States
    Competing interests
    The authors declare that no competing interests exist.
  13. Patrizia Camelliti

    School of Biosciences and Medicine, University of Surrey, Surrey, United Kingdom
    Competing interests
    The authors declare that no competing interests exist.
  14. Patrycja Dubielecka

    Brown University, Providence, United States
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0003-3987-0647
  15. Dmitry Terentyev

    Cardiovascular Research Center, Rhode Island Hospital, Providence, United States
    Competing interests
    The authors declare that no competing interests exist.
  16. Federica del Monte

    Medical University of South Carolina, Charleston, United States
    Competing interests
    The authors declare that no competing interests exist.
  17. Bum-Rak Choi

    Cardiovascular Research Center, Rhode Island Hospital, Providence, United States
    Competing interests
    The authors declare that no competing interests exist.
  18. John Sedivy

    Brown University, Providence, United States
    Competing interests
    The authors declare that no competing interests exist.
  19. Gideon Koren

    Brown University, Providence, United States
    For correspondence
    gideon_koren@brown.edu
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-6211-5837

Funding

NHLBI Division of Intramural Research (R01HL139467)

  • John Sedivy
  • Gideon Koren

NHLBI Division of Intramural Research (1R1AG049608-01)

  • John Sedivy
  • Gideon Koren

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

Ethics

Animal experimentation: This investigation conformed with the current Guide for Care and Use of Laboratory Animals published by the National Institutes of Health (NIH Publication, Revised 2011) as well as the standards recently delineated by the American Physiological Society ("Guiding Principles for Research Involving Animals and Human Beings") and was approved by the Institutional Animal Care and Use Committee of Rhode Island Hospital (Permits numbers 5001-21 and 5040-22).

Reviewing Editor

  1. Christopher L-H Huang, University of Cambridge, United Kingdom

Publication history

  1. Received: October 10, 2022
  2. Accepted: May 18, 2023
  3. Accepted Manuscript published: May 19, 2023 (version 1)

Copyright

© 2023, Baggett 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

  • 122
    Page views
  • 28
    Downloads
  • 0
    Citations

Article citation count generated by polling the highest count across the following sources: Crossref, PubMed Central, Scopus.

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. Brett C Baggett
  2. Kevin R Murphy
  3. Elif Sengun
  4. Eric Mi
  5. Yueming Cao
  6. Nilufer N Turan
  7. Yichun Lu
  8. Lorraine Scofield
  9. Tae Yun Kim
  10. Anatoli Y Kabakov
  11. Peter Bronk
  12. Zhilin Qu
  13. Patrizia Camelliti
  14. Patrycja Dubielecka
  15. Dmitry Terentyev
  16. Federica del Monte
  17. Bum-Rak Choi
  18. John Sedivy
  19. Gideon Koren
(2023)
Myofibroblast senescence promotes arrhythmogenic remodeling in the aged infarcted rabbit heart
eLife 12:e84088.
https://doi.org/10.7554/eLife.84088

Further reading

    1. Cell Biology
    Ignacio Bravo-Plaza, Victor G Tagua ... Miguel A Peñalva
    Research Article

    Uso1/p115 and RAB1 tether ER-derived vesicles to the Golgi. Uso1/p115 contains a globular-head-domain (GHD), a coiled-coil (CC) mediating dimerization/tethering and a C-terminal region (CTR) interacting with golgins. Uso1/p115 is recruited to vesicles by RAB1. Genetic studies placed Uso1 paradoxically acting upstream of, or in conjunction with RAB1 (Sapperstein et al., 1996). We selected two missense mutations in uso1 resulting in E6K and G540S in the GHD that rescued lethality of rab1-deficient Aspergillus nidulans. The mutations are phenotypically additive, their combination suppressing the complete absence of RAB1, which emphasizes the key physiological role of the GHD. In living hyphae Uso1 recurs on puncta (60 sec half-life) colocalizing partially with the Golgi markers RAB1, Sed5 and GeaA/Gea1/Gea2, and totally with the retrograde cargo receptor Rer1, consistent with Uso1 dwelling in a very early Golgi compartment from which ER residents reaching the Golgi recycled back to the ER. Localization of Uso1, but not of Uso1E6K/G540S, to puncta is abolished by compromising RAB1 function, indicating that E6K/G540S creates interactions bypassing RAB1. That Uso1 delocalization correlates with a decrease in the number of Gea1 cisternae supports that Uso1-and-Rer1-containing puncta are where the protein exerts its physiological role. In S-tag-coprecipitation experiments Uso1 is an associate of the Sed5/Bos1/Bet1/Sec22 SNARE complex zippering vesicles with the Golgi, with Uso1E6K/G540S showing stronger association. Using purified proteins, we show that Bos1 and Bet1 bind the Uso1 GHD directly. However, Bet1 is a strong E6K/G540S-independent binder, whereas Bos1 is weaker but becomes as strong as Bet1 when the GHD carries E6K/G540S. G540S alone markedly increases GHD binding to Bos1, whereas E6K causes a weaker effect, correlating with their phenotypic contributions. AlphaFold2 predicts that G540S increases binding of the GHD to the Bos1 Habc domain. In contrast, E6K lies in an N-terminal, potentially alpha-helical, region that sensitive genetic tests indicate as required for full Uso1 function. Remarkably, this region is at the end of the GHD basket opposite to the end predicted to interact with Bos1. We show that unlike dimeric full-length and CTR∆ Uso1 proteins, the GHD lacking the CC/CTR dimerization domain, whether originating from bacteria or Aspergillus extracts and irrespective of whether it carries or not E6K/G540S, would appear to be monomeric. With the finding that overexpression of E6K/G540S and wild-type GHD complement uso1∆, our data indicate that the GHD monomer is capable of providing, at least partially, the essential Uso1 functions, and that long-range tethering activity is dispensable. Rather, these findings strongly suggest that the essential role of Uso1 involves the regulation of SNAREs.

    1. Cell Biology
    Sandipan Dasgupta, Daniella Y Dayagi ... Jeffrey E Gerst
    Research Article

    Full-length mRNAs transfer between adjacent mammalian cells via direct cell-to-cell connections called tunneling nanotubes (TNTs). However, the extent of mRNA transfer at the transcriptome-wide level (the 'transferome') is unknown. Here, we analyzed the transferome in an in vitro human-mouse cell co-culture model using RNA-sequencing. We found that mRNA transfer is non-selective, prevalent across the human transcriptome, and that the amount of transfer to mouse embryonic fibroblasts (MEFs) strongly correlates with the endogenous level of gene expression in donor human breast cancer cells. Typically, <1% of endogenous mRNAs undergo transfer. Non-selective, expression-dependent RNA transfer was further validated using synthetic reporters. RNA transfer appears contact-dependent via TNTs, as exemplified for several mRNAs. Notably, significant differential changes in the native MEF transcriptome were observed in response to co-culture, including the upregulation of multiple cancer and cancer-associated fibroblast-related genes and pathways. Together, these results lead us to suggest that TNT-mediated RNA transfer could be a phenomenon of physiological importance under both normal and pathogenic conditions.