Activation of human aortic valve interstitial cells by local stiffness involves YAP-dependent transcriptional signaling

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

Differentiation of valve interstitial cells (VICs) into pro-calcific cells is one of the central events in calcific aortic valve (AoV) disease (CAVD). While the paracrine pathways and the responsivity of VICs to mechanical compliance of the surrounding environment are well characterized, the molecular programming related to variations in local stiffness, and its link to cytoskeleton dynamics, is less consolidated. By using a simple method to produce 2D poly-acrylamide gels with stiffness controlled with atomic force microscopy (AFM), we manufactured adhesion substrates onto which human VICs from stenotic valves were plated, and subsequently investigated for cytoskeleton dynamics and activation of the mechanosensing-related transcription factor YAP. As a comparison, we employed VICs from patients undergoing valve substitution for valve insufficiency, a non-calcific AoV disease, which does not involve extensive inflammation. While the two VICs types did not differ for basic responses onto substrates with different stiffness values (e.g. adhesion and proliferation), they were subject to a different dynamics of stiffness-dependent YAP nuclear shuttling, revealing for the first time an intracellular force transduction mechanism distinctive for calcific aortic valve disease. In VICs from stenotic valves, YAP nuclear translocation occurred in concert with an increase in cytoskeleton tensioning and loading of the myofibroblast-specific protein αSMA onto the F-actin cytoskeleton. AFM force mapping performed along radial sections of human calcific valve leaflets identified, finally, areas with high and low levels of rigidity within a similar range to those controlling YAP nuclear translocation in vitro. Since VICs juxtaposed to these areas exhibited nuclear localized YAP, we conclude that subtle variations in matrix stiffness are involved in mechanosensing-dependent VICs activation and pathological differentiation in CAVD.

Original languageEnglish
Pages (from-to)268-279
Number of pages12
JournalBiomaterials
Volume181
DOIs
Publication statusPublished - Oct 1 2018

Fingerprint

Aortic Valve
Chemical activation
Stiffness
Atomic force microscopy
Aortic Diseases
Adhesion
Cytoskeleton
Transcription factors
Stiffness matrix
Substrates
Atomic Force Microscopy
Polyacrylates
Rigidity
Actins
Substitution reactions
Transcription Factors
Gels
Proteins
Myofibroblasts
Acrylamide

Keywords

  • Calcific aortic valve disease
  • Mechanosensing
  • Tissue stiffness
  • Valve interstitial cells
  • Yes associated protein

ASJC Scopus subject areas

  • Bioengineering
  • Ceramics and Composites
  • Biophysics
  • Biomaterials
  • Mechanics of Materials

Cite this

@article{695ff390715f492b9ee206d7d9c08f25,
title = "Activation of human aortic valve interstitial cells by local stiffness involves YAP-dependent transcriptional signaling",
abstract = "Differentiation of valve interstitial cells (VICs) into pro-calcific cells is one of the central events in calcific aortic valve (AoV) disease (CAVD). While the paracrine pathways and the responsivity of VICs to mechanical compliance of the surrounding environment are well characterized, the molecular programming related to variations in local stiffness, and its link to cytoskeleton dynamics, is less consolidated. By using a simple method to produce 2D poly-acrylamide gels with stiffness controlled with atomic force microscopy (AFM), we manufactured adhesion substrates onto which human VICs from stenotic valves were plated, and subsequently investigated for cytoskeleton dynamics and activation of the mechanosensing-related transcription factor YAP. As a comparison, we employed VICs from patients undergoing valve substitution for valve insufficiency, a non-calcific AoV disease, which does not involve extensive inflammation. While the two VICs types did not differ for basic responses onto substrates with different stiffness values (e.g. adhesion and proliferation), they were subject to a different dynamics of stiffness-dependent YAP nuclear shuttling, revealing for the first time an intracellular force transduction mechanism distinctive for calcific aortic valve disease. In VICs from stenotic valves, YAP nuclear translocation occurred in concert with an increase in cytoskeleton tensioning and loading of the myofibroblast-specific protein αSMA onto the F-actin cytoskeleton. AFM force mapping performed along radial sections of human calcific valve leaflets identified, finally, areas with high and low levels of rigidity within a similar range to those controlling YAP nuclear translocation in vitro. Since VICs juxtaposed to these areas exhibited nuclear localized YAP, we conclude that subtle variations in matrix stiffness are involved in mechanosensing-dependent VICs activation and pathological differentiation in CAVD.",
keywords = "Calcific aortic valve disease, Mechanosensing, Tissue stiffness, Valve interstitial cells, Yes associated protein",
author = "Rosaria Santoro and Denis Scaini and Severino, {Luisa Ulloa} and Francesco Amadeo and Silvia Ferrari and Giacomo Bernava and Gloria Garoffolo and Marco Agrifoglio and Loredana Casalis and Maurizio Pesce",
year = "2018",
month = "10",
day = "1",
doi = "10.1016/j.biomaterials.2018.07.033",
language = "English",
volume = "181",
pages = "268--279",
journal = "Biomaterials",
issn = "0142-9612",
publisher = "Elsevier BV",

}

TY - JOUR

T1 - Activation of human aortic valve interstitial cells by local stiffness involves YAP-dependent transcriptional signaling

AU - Santoro, Rosaria

AU - Scaini, Denis

AU - Severino, Luisa Ulloa

AU - Amadeo, Francesco

AU - Ferrari, Silvia

AU - Bernava, Giacomo

AU - Garoffolo, Gloria

AU - Agrifoglio, Marco

AU - Casalis, Loredana

AU - Pesce, Maurizio

PY - 2018/10/1

Y1 - 2018/10/1

N2 - Differentiation of valve interstitial cells (VICs) into pro-calcific cells is one of the central events in calcific aortic valve (AoV) disease (CAVD). While the paracrine pathways and the responsivity of VICs to mechanical compliance of the surrounding environment are well characterized, the molecular programming related to variations in local stiffness, and its link to cytoskeleton dynamics, is less consolidated. By using a simple method to produce 2D poly-acrylamide gels with stiffness controlled with atomic force microscopy (AFM), we manufactured adhesion substrates onto which human VICs from stenotic valves were plated, and subsequently investigated for cytoskeleton dynamics and activation of the mechanosensing-related transcription factor YAP. As a comparison, we employed VICs from patients undergoing valve substitution for valve insufficiency, a non-calcific AoV disease, which does not involve extensive inflammation. While the two VICs types did not differ for basic responses onto substrates with different stiffness values (e.g. adhesion and proliferation), they were subject to a different dynamics of stiffness-dependent YAP nuclear shuttling, revealing for the first time an intracellular force transduction mechanism distinctive for calcific aortic valve disease. In VICs from stenotic valves, YAP nuclear translocation occurred in concert with an increase in cytoskeleton tensioning and loading of the myofibroblast-specific protein αSMA onto the F-actin cytoskeleton. AFM force mapping performed along radial sections of human calcific valve leaflets identified, finally, areas with high and low levels of rigidity within a similar range to those controlling YAP nuclear translocation in vitro. Since VICs juxtaposed to these areas exhibited nuclear localized YAP, we conclude that subtle variations in matrix stiffness are involved in mechanosensing-dependent VICs activation and pathological differentiation in CAVD.

AB - Differentiation of valve interstitial cells (VICs) into pro-calcific cells is one of the central events in calcific aortic valve (AoV) disease (CAVD). While the paracrine pathways and the responsivity of VICs to mechanical compliance of the surrounding environment are well characterized, the molecular programming related to variations in local stiffness, and its link to cytoskeleton dynamics, is less consolidated. By using a simple method to produce 2D poly-acrylamide gels with stiffness controlled with atomic force microscopy (AFM), we manufactured adhesion substrates onto which human VICs from stenotic valves were plated, and subsequently investigated for cytoskeleton dynamics and activation of the mechanosensing-related transcription factor YAP. As a comparison, we employed VICs from patients undergoing valve substitution for valve insufficiency, a non-calcific AoV disease, which does not involve extensive inflammation. While the two VICs types did not differ for basic responses onto substrates with different stiffness values (e.g. adhesion and proliferation), they were subject to a different dynamics of stiffness-dependent YAP nuclear shuttling, revealing for the first time an intracellular force transduction mechanism distinctive for calcific aortic valve disease. In VICs from stenotic valves, YAP nuclear translocation occurred in concert with an increase in cytoskeleton tensioning and loading of the myofibroblast-specific protein αSMA onto the F-actin cytoskeleton. AFM force mapping performed along radial sections of human calcific valve leaflets identified, finally, areas with high and low levels of rigidity within a similar range to those controlling YAP nuclear translocation in vitro. Since VICs juxtaposed to these areas exhibited nuclear localized YAP, we conclude that subtle variations in matrix stiffness are involved in mechanosensing-dependent VICs activation and pathological differentiation in CAVD.

KW - Calcific aortic valve disease

KW - Mechanosensing

KW - Tissue stiffness

KW - Valve interstitial cells

KW - Yes associated protein

UR - http://www.scopus.com/inward/record.url?scp=85051036298&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85051036298&partnerID=8YFLogxK

U2 - 10.1016/j.biomaterials.2018.07.033

DO - 10.1016/j.biomaterials.2018.07.033

M3 - Article

AN - SCOPUS:85051036298

VL - 181

SP - 268

EP - 279

JO - Biomaterials

JF - Biomaterials

SN - 0142-9612

ER -