Ablation of palladin in adult heart causes dilated cardiomyopathy associated with intercalated disc abnormalities

  1. Giuseppina Mastrototaro
  2. Pierluigi Carullo
  3. Jianlin Zhang
  4. Beatrice Scellini
  5. Nicoletta Piroddi
  6. Simona Nemska
  7. Maria Carmela Filomena
  8. Simone Serio
  9. Carol A Otey
  10. Chiara Tesi
  11. Fabian Emrich
  12. Wolfgang A Linke
  13. Corrado Poggesi
  14. Simona Boncompagni
  15. Marie-Louise Bang  Is a corresponding author
  1. IRCCS Humanitas Research Hospital, Italy
  2. Institute of Genetic and Biomedical Research, Italy
  3. University of California, San Diego, United States
  4. University of Florence, Italy
  5. University of North Carolina at Chapel Hill, United States
  6. Goethe University Hospital, Germany
  7. University of Münster, Germany
  8. University G d' Annunzio of Chieti, Italy

Abstract

Palladin (PALLD) belongs to the PALLD/myopalladin (MYPN)/myotilin family of actin-associated immunoglobulin-containing proteins in the sarcomeric Z-line. PALLD is ubiquitously expressed in several isoforms and its longest 200 kDa isoform, predominantly expressed in striated muscle, shows high structural homology to MYPN. MYPN gene mutations are associated with human cardiomyopathies, whereas the role of PALLD in the heart has remained unknown, partly due to embryonic lethality of PALLD knockout mice. In a yeast two-hybrid screening, CARP/Ankrd1 and FHOD1 were identified as novel interaction partners of PALLD's N-terminal region. To study the role of PALLD in the heart, we generated conditional (cPKO) and inducible (cPKOi) cardiomyocyte-specific PALLD knockout mice. While cPKO mice exhibited no pathological phenotype, ablation of PALLD in adult cPKOi mice caused progressive cardiac dilation and systolic dysfunction, associated with reduced cardiomyocyte contractility, intercalated disc abnormalities, and fibrosis, demonstrating that PALLD is essential for normal cardiac function. Double cPKO and MYPN knockout (MKO) mice exhibited a similar phenotype as MKO mice, suggesting that MYPN does not compensate for the loss of PALLD in cPKO mice. Altered transcript levels of MYPN and PALLD isoforms were found in myocardial tissue from human dilated and ischemic cardiomyopathy patients, whereas their protein expression levels were unaltered.

Data availability

All data generated and analysed during this study are included in the manuscript and figure supplements. Source Data files have been provided for all figures.

Article and author information

Author details

  1. Giuseppina Mastrototaro

    IRCCS Humanitas Research Hospital, Milan, Italy
    Competing interests
    The authors declare that no competing interests exist.
  2. Pierluigi Carullo

    Institute of Genetic and Biomedical Research, Milan, Italy
    Competing interests
    The authors declare that no competing interests exist.
  3. Jianlin Zhang

    School of Medicine, University of California, San Diego, San Diego, United States
    Competing interests
    The authors declare that no competing interests exist.
  4. Beatrice Scellini

    Department of Experimental and Clinical Medicine, University of Florence, Florence, Italy
    Competing interests
    The authors declare that no competing interests exist.
  5. Nicoletta Piroddi

    Department of Experimental and Clinical Medicine, University of Florence, Florence, Italy
    Competing interests
    The authors declare that no competing interests exist.
  6. Simona Nemska

    IRCCS Humanitas Research Hospital, Milan, Italy
    Competing interests
    The authors declare that no competing interests exist.
  7. Maria Carmela Filomena

    Institute of Genetic and Biomedical Research, Milan, Italy
    Competing interests
    The authors declare that no competing interests exist.
  8. Simone Serio

    IRCCS Humanitas Research Hospital, Milan, Italy
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-7294-2094
  9. Carol A Otey

    Department of Cell Biology and Physiology, University of North Carolina at Chapel Hill, Chapel Hill, United States
    Competing interests
    The authors declare that no competing interests exist.
  10. Chiara Tesi

    Department of Experimental and Clinical Medicine, University of Florence, Florence, Italy
    Competing interests
    The authors declare that no competing interests exist.
  11. Fabian Emrich

    Department of Cardiac Surgery, Goethe University Hospital, Frankfurt, Germany
    Competing interests
    The authors declare that no competing interests exist.
  12. Wolfgang A Linke

    Institute of Physiology II, University of Münster, Münster, Germany
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0003-0801-3773
  13. Corrado Poggesi

    Department of Experimental and Clinical Medicine, University of Florence, Florence, Italy
    Competing interests
    The authors declare that no competing interests exist.
  14. Simona Boncompagni

    Department of Neuroscience, Imaging and Clinical Sciences, University G d' Annunzio of Chieti, Chieti, Italy
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-5308-5069
  15. Marie-Louise Bang

    Institute of Genetic and Biomedical Research, Milan, Italy
    For correspondence
    marie-louise.bang@cnr.it
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-8859-5034

Funding

Fondazione Telethon (GGP12282)

  • Marie-Louise Bang

Ministero dell'Università e della Ricerca (2010R8JK2X_006)

  • Marie-Louise Bang

Horizon 2020 Framework Programme (777204)

  • Corrado Poggesi

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 animal studies were approved by the Italian Ministry of Health and performed in full compliance with the rules and regulations of the European Union (Directive 2010/63/EU of the European Parlia- ment) and Italy (Council of 22 September 2010; directive from the Italian Ministry of Health) on the protection of animals used for scientific purposes.

Human subjects: Human myocardial biopsies from cardiomyopathy patients were obtained from Leipzig Heart Center, Germany following approval by the institutional review board (protocol #240/16-ek) and signed informed consent from the patients according to the principles of the Declaration of Helsinki. Myocardial biopsies from healthy donors rejected for transplantation were obtained from Careggi University Hospital, Florence, Italy (protocol #2006/0024713; renewed May 2009).

Reviewing Editor

  1. Nuno Guimarães-Camboa, Goethe University Frankfurt, Germany

Publication history

  1. Received: March 14, 2022
  2. Accepted: March 15, 2023
  3. Accepted Manuscript published: March 16, 2023 (version 1)

Copyright

© 2023, Mastrototaro 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

  • 102
    Page views
  • 23
    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. Giuseppina Mastrototaro
  2. Pierluigi Carullo
  3. Jianlin Zhang
  4. Beatrice Scellini
  5. Nicoletta Piroddi
  6. Simona Nemska
  7. Maria Carmela Filomena
  8. Simone Serio
  9. Carol A Otey
  10. Chiara Tesi
  11. Fabian Emrich
  12. Wolfgang A Linke
  13. Corrado Poggesi
  14. Simona Boncompagni
  15. Marie-Louise Bang
(2023)
Ablation of palladin in adult heart causes dilated cardiomyopathy associated with intercalated disc abnormalities
eLife 12:e78629.
https://doi.org/10.7554/eLife.78629

Further reading

    1. Biochemistry and Chemical Biology
    2. Cancer Biology
    Mark Borris D Aldonza, Junghwa Cha ... Yoosik Kim
    Research Article

    Cancer secretome is a reservoir for aberrant glycosylation. How therapies alter this post- translational cancer hallmark and the consequences thereof remain elusive. Here we show that an elevated secretome fucosylation is a pan-cancer signature of both response and resistance to multiple targeted therapies. Large-scale pharmacogenomics revealed that fucosylation genes display widespread association with resistance to these therapies. In cancer cell cultures, xenograft mouse models, and patients, targeted kinase inhibitors distinctively induced core fucosylation of secreted proteins less than 60 kDa. Label-free proteomics of N-glycoproteomes identified fucosylation of the antioxidant PON1 as a critical component of the therapy-induced secretome (TIS). N-glycosylation of TIS and target core fucosylation of PON1 are mediated by the fucose salvage-FUT8-SLC35C1 axis with PON3 directly modulating GDP-Fuc transfer on PON1 scaffolds. Core fucosylation in the Golgi impacts PON1 stability and folding prior to secretion, promoting a more degradation-resistant PON1. Global and PON1-specific secretome de-N-glycosylation both limited the expansion of resistant clones in a tumor regression model. We defined the resistance-associated transcription factors (TFs) and genes modulated by the N-glycosylated TIS via a focused and transcriptome-wide analyses. These genes characterize the oxidative stress, inflammatory niche, and unfolded protein response as important factors for this modulation. Our findings demonstrate that core fucosylation is a common modification indirectly induced by targeted therapies that paradoxically promotes resistance.

    1. Biochemistry and Chemical Biology
    2. Developmental Biology
    Zengdi Zhang, Zan Huang ... Hai-Bin Ruan
    Research Article Updated

    In mammals, interactions between the bone marrow (BM) stroma and hematopoietic progenitors contribute to bone-BM homeostasis. Perinatal bone growth and ossification provide a microenvironment for the transition to definitive hematopoiesis; however, mechanisms and interactions orchestrating the development of skeletal and hematopoietic systems remain largely unknown. Here, we establish intracellular O-linked β-N-acetylglucosamine (O-GlcNAc) modification as a posttranslational switch that dictates the differentiation fate and niche function of early BM stromal cells (BMSCs). By modifying and activating RUNX2, O-GlcNAcylation promotes osteogenic differentiation of BMSCs and stromal IL-7 expression to support lymphopoiesis. In contrast, C/EBPβ-dependent marrow adipogenesis and expression of myelopoietic stem cell factor (SCF) is inhibited by O-GlcNAcylation. Ablating O-GlcNAc transferase (OGT) in BMSCs leads to impaired bone formation, increased marrow adiposity, as well as defective B-cell lymphopoiesis and myeloid overproduction in mice. Thus, the balance of osteogenic and adipogenic differentiation of BMSCs is determined by reciprocal O-GlcNAc regulation of transcription factors, which simultaneously shapes the hematopoietic niche.