Peri/epicellular protein disulfide isomerase-A1 acts as an upstream organizer of cytoskeletal mechanoadaptation in vascular smooth muscle cells

Autor(es):Mostrar menos -

Tanaka, Leonardo Y. ^[1] ; Araujo, Thais L. S. ^[1] ; Rodriguez, I, Andres ; Ferraz, Mariana S. ^[2] ; Pelegati, Vitor B. ^[3] ; Morais, Mauro C. C. ^{[4, 5, 6]} ; dos Santos, Aline M. ^[7] ; Cesar, Carlos L. ^[3] ; Ramos, Alexandre F. ^{[4, 5, 6]} ; Alencar, Adriano M. ^[2] ; Laurindo, Francisco R. M. ^[8]

Afiliação do(s) autor(es)

^[1] Univ Sao Paulo, Sch Med, Heart Inst, Vasc Biol Lab, Sao Paulo - Brazil

^[2] Univ Sao Paulo, Inst Phys, Sao Paulo - Brazil

^[3] Univ Estadual Campinas, Gleb Wataghin Inst Phys, Campinas, SP - Brazil

^[4] Univ Sao Paulo, Fac Med, Escola Artes Ciencias & Humanidades, Sao Paulo - Brazil

^[5] Univ Sao Paulo, Fac Med, Nucleo Estudos Interdisciplinares Sistemas Comple, Dept Radiol & Oncol, Sao Paulo - Brazil

^[6] Univ Sao Paulo, Fac Med, Ctr Pesquisa Translac Oncol, Inst Canc Estado Sao Paulo, Sao Paulo - Brazil

^[7] Univ Estadual Campinas, Inst Biol, Dept Struct & Funct Biol, Campinas, SP - Brazil

^[8] Rodriguez, Andres, I, Univ Sao Paulo, Sch Med, Heart Inst, Vasc Biol Lab, Sao Paulo - Brazil

• Tipo de documento: Artigo Científico
• Fonte: AMERICAN JOURNAL OF PHYSIOLOGY-HEART AND CIRCULATORY PHYSIOLOGY; v. 316, n. 3, p. H566-H579, MAR 2019.
• Citações Web of Science: 3

Resumo

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 beta(1)-integrin and promoted polarized compartmentalization of suifenylated 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 pecPDIA1 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 beta(1)-integrin and localized RhoA activation. pecPDIA1 acts as a novel organizer of mechanoadaptation responses in VSMCs. (AU)

• Processo FAPESP: 13/17115-5 - Dissulfeto-isomerase proteica epi/pericelular: um novo regulador de adaptação biomecânica em células vasculares
• Beneficiário: Leonardo Yuji Tanaka
• Modalidade de apoio: Bolsas no Brasil - Pós-Doutorado
• Processo FAPESP: 13/06241-0 - Modelo de interação entre dissulfeto-isomerase proteica e RhoGTPases e seu regulador RhoGDI durante a ativação do complexo NADPH oxidase em células musculares lisas
• Beneficiário: Andrés Ignacio Rodríguez Morales
• Modalidade de apoio: Bolsas no Brasil - Pós-Doutorado
• Processo FAPESP: 15/06210-2 - Envolvimento da via da GRASP na externalização da dissulfeto isomerase proteica-A1 e sua implicação na resposta de células endoteliais a shear stress
• Beneficiário: Thaís Larissa Araujo de Oliveira Silva
• Modalidade de apoio: Bolsas no Brasil - Pós-Doutorado
• Processo FAPESP: 13/07937-8 - Redoxoma
• Beneficiário: Ohara Augusto
• Modalidade de apoio: Auxílio à Pesquisa - Centros de Pesquisa, Inovação e Difusão - CEPIDs