Keloid fibroblasts have elevated and dysfunctional mechanotransduction signaling that is independent of TGF-β
| Autor: | Ian L. Chin, Yu Suk Choi, Fiona M. Wood, Manon Subilia, Mark W. Fear, Zhenjun Deng, Andrew W. Stevenson, Cecilia M Prêle, Nicole Hortin |
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| Rok vydání: | 2021 |
| Předmět: |
Adult
Male Adolescent Primary Cell Culture Dermatology Matrix (biology) Mechanotransduction Cellular Biochemistry Transforming Growth Factor beta1 Extracellular matrix Keloid Western blot medicine Humans Mechanotransduction Fibroblast Molecular Biology Cells Cultured Aged Skin medicine.diagnostic_test Chemistry YAP-Signaling Proteins Fibroblasts Middle Aged medicine.disease Phenotype Actins Cell biology medicine.anatomical_structure Female Signal Transduction Transforming growth factor |
| Zdroj: | Journal of Dermatological Science. 104:11-20 |
| ISSN: | 0923-1811 |
| Popis: | Background Fibroblasts found in keloid tissues are known to present an altered sensitivity to microenvironmental stimuli. However, the impact of changes in extracellular matrix stiffness on phenotypes of normal fibroblasts (NFs) and keloid fibroblasts (KFs) is poorly understood. Objectives Investigation the impact of matrix stiffness on NFs and KFs mainly via detecting yes-associated protein (YAP) expression. Methods We used fibronectin-coated polyacrylamide hydrogel substrates with a range from physiological to pathological stiffness values with or without TGF-β (fibrogenic inducer). Atomic force microscopy was used to measure the stiffness of fibroblasts. Cellular mechanoresponses were screened by immunocytochemistry, Western blot and Luminex assay. Results KFs are stiffer than NFs with greater expression of α-SMA. In NFs, YAP nuclear translocation was induced by increasing matrix stiffness as well as by stimulation with TGF-β. In contrast, KFs showed higher baseline levels of nuclear YAP that was not responsive to matrix stiffness or TGF-β. TGF-β1 induced p-SMAD3 in both KFs and NFs, demonstrating the pathway was functional and not hyperactivated in KFs. Moreover, blebbistatin suppressed α-SMA expression and cellular stiffness in KFs, linking the elevated YAP signaling to keloid phenotype. Conclusions These data suggest that whilst normal skin fibroblasts respond to matrix stiffness in vitro, keloid fibroblasts have elevated activation of mechanotransduction signaling insensitive to the microenvironment. This elevated signaling appears linked to the expression of α-SMA, suggesting a direct link to disease pathogenesis. These findings suggest changes to keloid fibroblast phenotype related to mechanotransduction contribute to disease and may be a useful therapeutic target. |
| Databáze: | OpenAIRE |
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