Sialylation regulates myofibroblast differentiation of human skin fibroblasts

Abstract

BackgroundFibroblasts are key players in maintaining skin homeostasis and in orchestrating physiological tissue repair and skin regeneration. Dysfunctions in fibroblasts that occur with aging and the senescent process lead to the delayed healing observed in elderly people. The molecular mechanisms leading to fibroblast dysfunction during aging and the senescent process have not yet been clarified. Previously, changes in patterns of glycosylation were observed in fibroblasts in aging and the senescent process, but the effect of these changes on the function of fibroblasts has not been well documented. Here, we investigated whether changes in glycosylation during the process to senescence may have functional effects on fibroblasts.MethodsThe changes in cell surface glycans on skin fibroblasts during the process to senescence were examined in early-passage (EP) and late-passage (LP) skin fibroblasts by fluorescence-activated cell sorting analysis using lectins. The contributors to the changes in cell surface glycans were examined by real-time polymerase chain reaction or Western blot analysis. The effects of changes in glycosylation on proliferation, migration, induction of cellular senescence, and myofibroblast differentiation induced by transforming growth factor (TGF)-β1 stimulation were examined in EP fibroblasts. The changes in glycosylation were performed by GalNAc-α-O-benzyl or sialidase treatment.ResultsA decrease in sialylation of glycoproteins and an increase in sialidase NEU1 were observed in LP fibroblasts. The reduction of sialylation did not have any effect on proliferation, migration, or induction of cellular senescence. On the other hand, myofibroblast differentiation was inhibited by the reduction of sialylation, indicating that sialylation is important for myofibroblast differentiation. The localization of CD44 in lipid rafts, which is required for myofibroblast differentiation, was inhibited by the reduction of sialylation. Furthermore, reduced myofibroblast differentiation in LP fibroblasts was restored by a sialidase inhibitor.ConclusionsDesialylation of CD44 with increased sialidase during the process to senescence reduced the localization of CD44 in lipid rafts after TGF-β1 stimulation, leading to the inhibition of myofibroblast differentiation. Thus, regulation of sialylation may be an attractive strategy for the prevention and regenerative therapy of age-related skin diseases, cosmetic skin alterations, and chronic wounds caused by delayed healing in elderly people.Electronic supplementary materialThe online version of this article (doi:10.1186/s13287-017-0534-1) contains supplementary material, which is available to authorized users.

Establishment and characterization of a novel neuroendocrine carcinoma cell line derived from a human ascending colon tumor

“…Cells were harvested and dissociated as previously described using Accutase cell detachment solution (Merck Millipore). Dissociated single cells were incubated with rabbit polyclonal anti‐human CD133 (ab19898; Abcam) diluted in FACS buffer (0.5% [w/v] BSA and 0.1% [w/v] sodium azide in PBS) for 30 minutes on ice.…”

Increase of MAL-II Binding Alpha2,3-Sialylated Glycan Is Associated with 5-FU Resistance and Short Survival of Cholangiocarcinoma Patients

“…Sialylation is a process of adding a terminal sialic acid (Sia) to the subterminal monosaccharide of carbohydrate chains on glycoproteins or glycolipids. It plays many roles in normal physiology, for example, neural differentiation [6], tissue regeneration [7,8], and resistance to influenza virus infection [9,10]. The increase of sialylation and sialylated-glycans were associated with development and progression of human diseases including cancer [10][11][12][13][14][15].…”

Glycosylation of proteins of human skin fibroblasts is changed by rosmarinic acid

“…Carbohydrate antigens are widely expressed on the cell surfaces and in extracellular matrix (ECM) as components of different glycoproteins. They contribute significantly to many crucial, biological functions, such as cell differentiation, cell adhesion, cell-cell interactions, and regulation of signaling pathways (Sasaki et al 2017). Glycans of the human cells are mostly linked to serine/threonine or asparagine of polypeptide chains forming O-or N-glycoproteins.…”

“…Fibroblasts are essential in maintaining skin homeostasis and participate in physiological tissue repair and skin regeneration. It has been demonstrated that cell surface glycans on fibroblasts can be affected by different external factors, e.g., during aging (Sasaki et al 2017;Itakura et al 2016).…”