| Autor: |
Tanaka LY; Vascular Biology Laboratory, Heart Institute, University of São Paulo School of Medicine , São Paulo , Brazil., Araujo TLS; Vascular Biology Laboratory, Heart Institute, University of São Paulo School of Medicine , São Paulo , Brazil., Rodriguez AI; Vascular Biology Laboratory, Heart Institute, University of São Paulo School of Medicine , São Paulo , Brazil.; Group of Research and Innovation in Vascular Health, Department of Basic Sciences, Faculty of Sciences, University of Bío-Bío , Chillán , Chile., Ferraz MS; Institute of Physics, University of São Paulo , São Paulo , Brazil., Pelegati VB; 'Gleb Wataghin' Institute of Physics, University of Campinas , Campinas , Brazil., Morais MCC; Escola de Artes, Ciências e Humanidades e Núcleo de Estudos Interdisciplinares em Sistemas Complexos, Departamento de Radiologia e Oncologia e Centro de Pesquisa Translacional em Oncologia - Instituto do Cancer do Estado São Paulo, Faculdade de Medicina, Universidade de São Paulo , São Paulo , Brazil., Santos AMD; Department of Structural and Functional Biology, Institute of Biology, University of Campinas , Campinas , Brazil., Cesar CL; 'Gleb Wataghin' Institute of Physics, University of Campinas , Campinas , Brazil., Ramos AF; Escola de Artes, Ciências e Humanidades e Núcleo de Estudos Interdisciplinares em Sistemas Complexos, Departamento de Radiologia e Oncologia e Centro de Pesquisa Translacional em Oncologia - Instituto do Cancer do Estado São Paulo, Faculdade de Medicina, Universidade de São Paulo , São Paulo , Brazil., Alencar AM; Institute of Physics, University of São Paulo , São Paulo , Brazil., Laurindo FRM; Vascular Biology Laboratory, Heart Institute, University of São Paulo School of Medicine , São Paulo , Brazil. |
| Abstrakt: |
Although redox processes closely interplay with mechanoresponses to control vascular remodeling, redox pathways coupling mechanostimulation to cellular cytoskeletal organization remain unclear. The peri/epicellular pool of protein disulfide isomerase-A1 (pecPDIA1) supports postinjury vessel remodeling. Using distinct models, we investigated whether pecPDIA1 could work as a redox-dependent organizer of cytoskeletal mechanoresponses. In vascular smooth muscle cells (VSMCs), pecPDIA1 immunoneutralization impaired stress fiber assembly in response to equibiaxial stretch and, under uniaxial stretch, significantly perturbed cell repositioning perpendicularly to stretch orientation. During cyclic stretch, pecPDIA1 supported thiol oxidation of the known mechanosensor β 1 -integrin and promoted polarized compartmentalization of sulfenylated proteins. Using traction force microscopy, we showed that pecPDIA1 organizes intracellular force distribution. The net contractile moment ratio of platelet-derived growth factor-exposed to basal VSMCs decreased from 0.90 ± 0.09 (IgG-exposed controls) to 0.70 ± 0.08 after pecPDI neutralization ( P < 0.05), together with an enhanced coefficient of variation for distribution of force modules, suggesting increased noise. Moreover, in a single cell model, pecPDIA1 neutralization impaired migration persistence without affecting total distance or velocity, whereas siRNA-mediated total PDIA1 silencing disabled all such variables of VSMC migration. Neither expression nor total activity of the master mechanotransmitter/regulator RhoA was affected by pecPDIA1 neutralization. However, cyclic stretch-induced focal distribution of membrane-bound RhoA was disrupted by pecPDI inhibition, which promoted a nonpolarized pattern of RhoA/caveolin-3 cluster colocalization. Accordingly, FRET biosensors showed that pecPDIA1 supports localized RhoA activity at cell protrusions versus perinuclear regions. Thus, pecPDI acts as a thiol redox-dependent organizer and noise reducer mechanism of cytoskeletal repositioning, oxidant generation, and localized RhoA activation during a variety of VSMC mechanoresponses. NEW & NOTEWORTHY Effects of a peri/epicellular pool of protein disulfide isomerase-A1 (pecPDIA1) during mechanoregulation in vascular smooth muscle cells (VSMCs) were highlighted using approaches such as equibiaxial and uniaxial stretch, random single cell migration, and traction force microscopy. pecPDIA1 regulates organization of the cytoskeleton and minimizes the noise of cell alignment, migration directionality, and persistence. pecPDIA1 mechanisms involve redox control of β 1 -integrin and localized RhoA activation. pecPDIA1 acts as a novel organizer of mechanoadaptation responses in VSMCs. |
