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Truncation of the N-terminus of cardiac troponin I initiates adaptive remodeling of the myocardial proteosome via phosphorylation of mechano-sensitive signaling pathways

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Abstract

The cardiac isoform of troponin I has a unique N-terminal extension (~ 1–30 amino acids), which contributes to the modulation of cardiac contraction and relaxation. Hearts of various species including humans produce a truncated variant of cardiac troponin I (cTnI-ND) deleting the first ~ 30 amino acids as an adaption in pathophysiological conditions. In this study, we investigated the impact of cTnI-ND chronic expression in transgenic mouse hearts compared to wildtype (WT) controls (biological n = 8 in each group). We aimed to determine the global phosphorylation effects of cTnI-ND on the cardiac proteome, thereby determining the signaling pathways that have an impact on cardiac function. The samples were digested and isobarically labeled and equally mixed for relative quantification via nanoLC-MS/MS. The peptides were then enriched for phospho-peptides and bioinformatic analysis was done with Ingenuity Pathway Analysis (IPA). We found approximately 77% replacement of the endogenous intact cTnI with cTnI-ND in the transgenic mouse hearts with 1674 phospho-proteins and 2971 non-modified proteins. There were 73 significantly altered phospho-proteins; bioinformatic analysis identified the top canonical pathways as associated with integrin, protein kinase A, RhoA, and actin cytoskeleton signaling. Among the 73 phospho-proteins compared to controls cTnI-ND hearts demonstrated a significant decrease in paxillin and YAP1, which are known to play a role in cell mechano-sensing pathways. Our data indicate that cTnI-ND modifications in the sarcomere are sufficient to initiate changes in the phospho-signaling profile that may underly the chronic-adaptive response associated with cTnI cleavage in response to stressors by modifying mechano-sensitive signaling pathways.

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Data availability

All data are contained within the article, supplementary data, or available from the corresponding author R. John Solaro, email: solarorj@uic.edu. The raw mass spectrometry data have been deposited in the ProteomeXchange Consortium (http://proteomecentral.proteomexchange.org) via the MassIVE partner repository (https://massive.ucsd.edu/ProteoSAFe/static/massive.jsp) with the dataset identifier MSV000087049.

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Acknowledgements

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Funding

This work was supported by NIH grants RO1 HL128468 (to BMW and RJS), PO1 HL062426 Project 1, 3, and Core C (to RJS, PPdT, CMW), and RO1 HL127691 (to JPJ, RJS, and PPdT). The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health.

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Authors and Affiliations

  1. Department of Physiology & Biophysics, Center for Cardiovascular Research, University of Illinois at Chicago, Chicago, USA

    Chad M. Warren, Monika Halas, Paul H. Goldspink, Han-Zhong Feng, Beata M. Wolska, Pieter P. de Tombe & Jian-Ping Jin

  2. Department of Physiology, Wayne State University School of Medicine, Detroit, MI, USA

    Han-Zhong Feng & Jian-Ping Jin

  3. UC Davis Genome Center, Davis, CA, USA

    Anthony W. Herren

  4. Division of Cardiology, Department of Medicine, University of Illinois at Chicago, Chicago, USA

    Beata M. Wolska

  5. Phymedexp, Université de Montpellier, Inserm, CNRS, Montpellier, France

    Pieter P. de Tombe

  6. Department of Physiology & Biophysics, College of Medicine, Center for Cardiovascular Research, University of Illinois at Chicago, 835 South Wolcott Avenue RM. E-202 (M/C 901), Chicago, IL, 60612, USA

    R. John Solaro

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Contributions

CMW contributed to writing of the original draft, writing, reviewing, & editing of the manuscript, methodology, investigation, validation, and formal analysis; MH contributed to investigation, formal analysis, and writing, reviewing, & editing of the manuscript; PHG contributed to writing of the original draft and writing, reviewing, & editing of the manuscript; HZF contributed to resources and writing, reviewing, & editing of the manuscript; AWH contributed to resources, formal analysis, and writing, reviewing, & editing of the manuscript; BMW contributed to writing, reviewing, & editing of the manuscript; PPdT contributed to writing, reviewing, & editing of the manuscript and funding acquisition. JPJ conceptualized and contributed to funding acquisition and writing, reviewing, & editing of the manuscript; RJS contributed to writing of the original draft, writing, reviewing, & editing of the manuscript, conceptualization, and funding acquisition.

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Correspondence to R. John Solaro.

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Conflict of interest

R. John Solaro is a member of the Scientific Advisory Board of Cytokinetics, Inc., a consultant to Pfizer, Inc., and a member of the Heart Failure Advisory Board of Amgen. The authors declare no additional competing financial interests.

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This article contains studies with animals and all animal protocols were approved by the animal care and use committee of the University of Illinois at Chicago, which also conforms to the “Guide for the Care and Use of Laboratory Animals” published by the United States National Institutes of Health, 8th edition, revised 2011.

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Warren, C.M., Halas, M., Goldspink, P.H. et al. Truncation of the N-terminus of cardiac troponin I initiates adaptive remodeling of the myocardial proteosome via phosphorylation of mechano-sensitive signaling pathways. Mol Cell Biochem 477, 1803–1815 (2022). https://doi.org/10.1007/s11010-022-04414-3

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