Fibrotic extracellular matrix impacts cardiomyocyte phenotype and function in an iPSC-derived isogenic model of cardiac fibrosis

Francesco Niro; Soraia Fernandes; Marco Cassani; Monica Apostolico; Jorge Oliver-De La Cruz; Daniel Pereira-Sousa; Stefania Pagliari; Vladimir Vinarsky; Zbyněk Zdráhal; David Potesil; Vaclav Pustka; Giulio Pompilio; Elena Sommariva; Davide Rovina; Angela Serena Maione; Luca Bersanini; Malin Becker; Marco Rasponi; Giancarlo Forte

doi:10.1016/j.trsl.2024.07.003

Fibrotic extracellular matrix impacts cardiomyocyte phenotype and function in an iPSC-derived isogenic model of cardiac fibrosis

Francesco Niro, Soraia Fernandes, Marco Cassani, Monica Apostolico, Jorge Oliver-De La Cruz, Daniel Pereira-Sousa, Stefania Pagliari, Vladimir Vinarsky, Zbyněk Zdráhal, David Potesil, Vaclav Pustka, Giulio Pompilio, Elena Sommariva, Davide Rovina, Angela Serena Maione, Luca Bersanini, Malin Becker, Marco Rasponi, Giancarlo Forte^*

^*Corresponding author for this work

SCMMS Denmark Hill

Research output: Contribution to journal › Article › peer-review

3 Citations (Scopus)

Abstract

Cardiac fibrosis occurs following insults to the myocardium and is characterized by the abnormal accumulation of non-compliant extracellular matrix (ECM), which compromises cardiomyocyte contractile activity and eventually leads to heart failure. This phenomenon is driven by the activation of cardiac fibroblasts (cFbs) to myofibroblasts and results in changes in ECM biochemical, structural and mechanical properties. The lack of predictive in vitro models of heart fibrosis has so far hampered the search for innovative treatments, as most of the cellular-based in vitro reductionist models do not take into account the leading role of ECM cues in driving the progression of the pathology. Here, we devised a single-step decellularization protocol to obtain and thoroughly characterize the biochemical and micro-mechanical properties of the ECM secreted by activated cFbs differentiated from human induced pluripotent stem cells (iPSCs). We activated iPSC-derived cFbs to the myofibroblast phenotype by tuning basic fibroblast growth factor (bFGF) and transforming growth factor beta 1 (TGF-β1) signalling and confirmed that activated cells acquired key features of myofibroblast phenotype, like SMAD2/3 nuclear shuttling, the formation of aligned alpha-smooth muscle actin (α−SMA)-rich stress fibres and increased focal adhesions (FAs) assembly. Next, we used Mass Spectrometry, nanoindentation, scanning electron and confocal microscopy to unveil the characteristic composition and the visco-elastic properties of the abundant, collagen-rich ECM deposited by cardiac myofibroblasts in vitro. Finally, we demonstrated that the fibrotic ECM activates mechanosensitive pathways in iPSC-derived cardiomyocytes, impacting on their shape, sarcomere assembly, phenotype, and calcium handling properties. We thus propose human bio-inspired decellularized matrices as animal-free, isogenic cardiomyocyte culture substrates recapitulating key pathophysiological changes occurring at the cellular level during cardiac fibrosis.

Original language	English
Pages (from-to)	58-77
Number of pages	20
Journal	Translational Research
Volume	273
DOIs	https://doi.org/10.1016/j.trsl.2024.07.003
Publication status	Published - Nov 2024

Keywords

Cardiac fibrosis modelling
Decellularized extracellular matrix
Induced pluripotent stem cells
iPSC-derived-cardiac fibroblasts
iPSC-derived-cardiomyocytes

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

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10.1016/j.trsl.2024.07.003

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Niro, F., Fernandes, S., Cassani, M., Apostolico, M., Oliver-De La Cruz, J., Pereira-Sousa, D., Pagliari, S., Vinarsky, V., Zdráhal, Z., Potesil, D., Pustka, V., Pompilio, G., Sommariva, E., Rovina, D., Maione, A. S., Bersanini, L., Becker, M., Rasponi, M., & Forte, G. (2024). Fibrotic extracellular matrix impacts cardiomyocyte phenotype and function in an iPSC-derived isogenic model of cardiac fibrosis. Translational Research, 273, 58-77. https://doi.org/10.1016/j.trsl.2024.07.003

@article{0b3e7082ae3b410688bfe6b0bd72678a,

title = "Fibrotic extracellular matrix impacts cardiomyocyte phenotype and function in an iPSC-derived isogenic model of cardiac fibrosis",

abstract = "Cardiac fibrosis occurs following insults to the myocardium and is characterized by the abnormal accumulation of non-compliant extracellular matrix (ECM), which compromises cardiomyocyte contractile activity and eventually leads to heart failure. This phenomenon is driven by the activation of cardiac fibroblasts (cFbs) to myofibroblasts and results in changes in ECM biochemical, structural and mechanical properties. The lack of predictive in vitro models of heart fibrosis has so far hampered the search for innovative treatments, as most of the cellular-based in vitro reductionist models do not take into account the leading role of ECM cues in driving the progression of the pathology. Here, we devised a single-step decellularization protocol to obtain and thoroughly characterize the biochemical and micro-mechanical properties of the ECM secreted by activated cFbs differentiated from human induced pluripotent stem cells (iPSCs). We activated iPSC-derived cFbs to the myofibroblast phenotype by tuning basic fibroblast growth factor (bFGF) and transforming growth factor beta 1 (TGF-β1) signalling and confirmed that activated cells acquired key features of myofibroblast phenotype, like SMAD2/3 nuclear shuttling, the formation of aligned alpha-smooth muscle actin (α−SMA)-rich stress fibres and increased focal adhesions (FAs) assembly. Next, we used Mass Spectrometry, nanoindentation, scanning electron and confocal microscopy to unveil the characteristic composition and the visco-elastic properties of the abundant, collagen-rich ECM deposited by cardiac myofibroblasts in vitro. Finally, we demonstrated that the fibrotic ECM activates mechanosensitive pathways in iPSC-derived cardiomyocytes, impacting on their shape, sarcomere assembly, phenotype, and calcium handling properties. We thus propose human bio-inspired decellularized matrices as animal-free, isogenic cardiomyocyte culture substrates recapitulating key pathophysiological changes occurring at the cellular level during cardiac fibrosis.",

keywords = "Cardiac fibrosis modelling, Decellularized extracellular matrix, Induced pluripotent stem cells, iPSC-derived-cardiac fibroblasts, iPSC-derived-cardiomyocytes",

author = "Francesco Niro and Soraia Fernandes and Marco Cassani and Monica Apostolico and {Oliver-De La Cruz}, Jorge and Daniel Pereira-Sousa and Stefania Pagliari and Vladimir Vinarsky and Zbyn{\v e}k Zdr{\'a}hal and David Potesil and Vaclav Pustka and Giulio Pompilio and Elena Sommariva and Davide Rovina and Maione, {Angela Serena} and Luca Bersanini and Malin Becker and Marco Rasponi and Giancarlo Forte",

note = "Publisher Copyright: {\textcopyright} 2024 The Author(s)",

year = "2024",

month = nov,

doi = "10.1016/j.trsl.2024.07.003",

language = "English",

volume = "273",

pages = "58--77",

journal = "Translational Research",

issn = "1931-5244",

publisher = "Elsevier",

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Niro, F, Fernandes, S, Cassani, M, Apostolico, M, Oliver-De La Cruz, J, Pereira-Sousa, D, Pagliari, S, Vinarsky, V, Zdráhal, Z, Potesil, D, Pustka, V, Pompilio, G, Sommariva, E, Rovina, D, Maione, AS, Bersanini, L, Becker, M, Rasponi, M & Forte, G 2024, 'Fibrotic extracellular matrix impacts cardiomyocyte phenotype and function in an iPSC-derived isogenic model of cardiac fibrosis', Translational Research, vol. 273, pp. 58-77. https://doi.org/10.1016/j.trsl.2024.07.003

TY - JOUR

T1 - Fibrotic extracellular matrix impacts cardiomyocyte phenotype and function in an iPSC-derived isogenic model of cardiac fibrosis

AU - Niro, Francesco

AU - Fernandes, Soraia

AU - Cassani, Marco

AU - Apostolico, Monica

AU - Oliver-De La Cruz, Jorge

AU - Pereira-Sousa, Daniel

AU - Pagliari, Stefania

AU - Vinarsky, Vladimir

AU - Zdráhal, Zbyněk

AU - Potesil, David

AU - Pustka, Vaclav

AU - Pompilio, Giulio

AU - Sommariva, Elena

AU - Rovina, Davide

AU - Maione, Angela Serena

AU - Bersanini, Luca

AU - Becker, Malin

AU - Rasponi, Marco

AU - Forte, Giancarlo

PY - 2024/11

Y1 - 2024/11

N2 - Cardiac fibrosis occurs following insults to the myocardium and is characterized by the abnormal accumulation of non-compliant extracellular matrix (ECM), which compromises cardiomyocyte contractile activity and eventually leads to heart failure. This phenomenon is driven by the activation of cardiac fibroblasts (cFbs) to myofibroblasts and results in changes in ECM biochemical, structural and mechanical properties. The lack of predictive in vitro models of heart fibrosis has so far hampered the search for innovative treatments, as most of the cellular-based in vitro reductionist models do not take into account the leading role of ECM cues in driving the progression of the pathology. Here, we devised a single-step decellularization protocol to obtain and thoroughly characterize the biochemical and micro-mechanical properties of the ECM secreted by activated cFbs differentiated from human induced pluripotent stem cells (iPSCs). We activated iPSC-derived cFbs to the myofibroblast phenotype by tuning basic fibroblast growth factor (bFGF) and transforming growth factor beta 1 (TGF-β1) signalling and confirmed that activated cells acquired key features of myofibroblast phenotype, like SMAD2/3 nuclear shuttling, the formation of aligned alpha-smooth muscle actin (α−SMA)-rich stress fibres and increased focal adhesions (FAs) assembly. Next, we used Mass Spectrometry, nanoindentation, scanning electron and confocal microscopy to unveil the characteristic composition and the visco-elastic properties of the abundant, collagen-rich ECM deposited by cardiac myofibroblasts in vitro. Finally, we demonstrated that the fibrotic ECM activates mechanosensitive pathways in iPSC-derived cardiomyocytes, impacting on their shape, sarcomere assembly, phenotype, and calcium handling properties. We thus propose human bio-inspired decellularized matrices as animal-free, isogenic cardiomyocyte culture substrates recapitulating key pathophysiological changes occurring at the cellular level during cardiac fibrosis.

AB - Cardiac fibrosis occurs following insults to the myocardium and is characterized by the abnormal accumulation of non-compliant extracellular matrix (ECM), which compromises cardiomyocyte contractile activity and eventually leads to heart failure. This phenomenon is driven by the activation of cardiac fibroblasts (cFbs) to myofibroblasts and results in changes in ECM biochemical, structural and mechanical properties. The lack of predictive in vitro models of heart fibrosis has so far hampered the search for innovative treatments, as most of the cellular-based in vitro reductionist models do not take into account the leading role of ECM cues in driving the progression of the pathology. Here, we devised a single-step decellularization protocol to obtain and thoroughly characterize the biochemical and micro-mechanical properties of the ECM secreted by activated cFbs differentiated from human induced pluripotent stem cells (iPSCs). We activated iPSC-derived cFbs to the myofibroblast phenotype by tuning basic fibroblast growth factor (bFGF) and transforming growth factor beta 1 (TGF-β1) signalling and confirmed that activated cells acquired key features of myofibroblast phenotype, like SMAD2/3 nuclear shuttling, the formation of aligned alpha-smooth muscle actin (α−SMA)-rich stress fibres and increased focal adhesions (FAs) assembly. Next, we used Mass Spectrometry, nanoindentation, scanning electron and confocal microscopy to unveil the characteristic composition and the visco-elastic properties of the abundant, collagen-rich ECM deposited by cardiac myofibroblasts in vitro. Finally, we demonstrated that the fibrotic ECM activates mechanosensitive pathways in iPSC-derived cardiomyocytes, impacting on their shape, sarcomere assembly, phenotype, and calcium handling properties. We thus propose human bio-inspired decellularized matrices as animal-free, isogenic cardiomyocyte culture substrates recapitulating key pathophysiological changes occurring at the cellular level during cardiac fibrosis.

KW - Cardiac fibrosis modelling

KW - Decellularized extracellular matrix

KW - Induced pluripotent stem cells

KW - iPSC-derived-cardiac fibroblasts

KW - iPSC-derived-cardiomyocytes

UR - http://www.scopus.com/inward/record.url?scp=85199087451&partnerID=8YFLogxK

U2 - 10.1016/j.trsl.2024.07.003

DO - 10.1016/j.trsl.2024.07.003

M3 - Article

C2 - 39025226

AN - SCOPUS:85199087451

SN - 1931-5244

VL - 273

SP - 58

EP - 77

JO - Translational Research

JF - Translational Research

ER -

Fibrotic extracellular matrix impacts cardiomyocyte phenotype and function in an iPSC-derived isogenic model of cardiac fibrosis

Abstract

Keywords

UN SDGs

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