Background: Wound healing and scarring are driven by transforming growth factor-␤1 (TGF-␤1)-dependent fibroblast to myofibroblast differentiation. Results: Cell surface CD44 and epidermal growth factor receptor (EGFR) co-localize in lipid rafts to signal through mitogenactivated protein kinase 1/2 (ERK1/2) and Ca 2ϩ /calmodulin kinase II (CaMKII). Conclusion: CD44 moves into lipid rafts in a TGF-␤1-and hyaluronan-dependent manner, co-localizes with EGFR, and triggers differentiation. Significance: This pathway presents novel targets for the therapy of wound-healing and fibrosis.

This study aimed to understand the role of the matrix polysaccharide, hyaluronan (HA), in influencing the scarring process by assessing its impact on regulating fibroblast behavior. Donormatched human oral and dermal fibroblasts were used as models of nonscarring and scarring fibroblast phenotypes, respectively. Phenotypic differences in these two fibroblast populations were assessed and related to differences in HA synthesis and assembly. The two fibroblast populations showed intrinsic differences in their response to the profibrotic cytokine, transforming growth factor-␤ 1 (TGF␤ 1 ), in that oral fibroblasts were resistant to TGF␤ 1 -driven myofibroblastic differentiation. In dermal fibroblasts, differentiation was associated with an induction of HA synthase (HAS1 and HAS2) transcription and assembly of pericellular HA coats. In comparison, resistance to differentiation in oral fibroblasts was associated with failure of induction of HAS1 and HAS2 transcription and failure of pericellular coat assembly. Furthermore, inhibition of HA synthesis in dermal fibroblasts significantly attenuated TGF␤ 1 -mediated differentiation. Interleukin-1␤ stimulation resulted in induction of HAS1 and HAS2 transcription but did not induce phenotypic differentiation or induce HA coat assembly. In addition, neither overexpression nor down-regulation of HAS1 (the isoform uniquely deficient in nonscarring oral fibroblasts) influenced phenotypic differentiation. In conclusion, inhibiting HA synthesis modulates TGF␤ 1 -dependent responses in these cells preventing fibroblast to myofibroblast differentiation. Moreover, HA pericellular coat assembly, rather than HAS isoform expression, appears to be associated with phenotypic differentiation.

No abstract

MicroRNAs are short noncoding RNA regulators that repress synthesis of their targets post-transcriptionally. On average, each microRNA is estimated to regulate several hundred protein-coding genes, and about 60% of proteins are thought to be regulated by microRNAs in total. A subset of these genes, including the key profibrotic cytokine transforming growth factor beta-1 (TGF-β1), exhibits particularly strong levels of post-transcriptional control of protein synthesis, involving microRNAs and other mechanisms. Changes in microRNA expression pattern are linked to profound effects on cell phenotype, and microRNAs have an emerging role in diverse physiological and pathological processes. In this review, we provide an overview of microRNA biology with a focus on their emerging role in diseases typified by organ fibrosis.

Background: Intracellular proteins glycosylation with O-GlcNAc is able to influence cell microenvironment. Results: O-GlcNAcylation increases hyaluronan synthase 2 (HAS2) transcription via its natural antisense transcript HAS2-AS1. Conclusion: A novel mechanism to regulate hyaluronan synthesis via long non-coding RNA is described. Significance: This finding highlights a new target to regulate HA synthesis, critical in many pathophysiological processes.

No abstract

We have previously demonstrated that transforming growth factor-beta1 (TGF-beta1)-mediated fibroblast-myofibroblast differentiation is associated with accumulation of a hyaluronan (HA) pericellular coat. The current study demonstrates failure of fibroblast-myofibroblast differentiation associated with in vitro aging. This is associated with attenuation of numerous TGF-beta1-dependent responses, including HA synthesis and induction of the HA synthase enzyme HAS2 and the hyaladherin tumor necrosis factor-alpha-stimulated gene 6 (TSG-6), which led to an age-related defect in pericellular HA coat assembly. Inhibition of HAS2-dependent HA synthesis by gene silencing, removal of the HA coat by hyaluronidase digestion, or gene silencing of TSG-6 or cell surface receptor CD44 led to abrogation of TGF-beta1-dependent induction of alpha-smooth muscle actin in "young" cells. This result supports the importance of HAS2-dependent HA synthesis and the HA coat during phenotypic activation. Interleukin-1beta stimulation, however, failed to promote phenotypic conversion despite coat formation. A return to basal levels of HA synthesis in aged cells by HAS2 overexpression restored TGF-beta1-dependent induction of TSG-6 and pericellular HA coat assembly. However, this did not lead to the acquisition of a myofibroblast phenotype. Coordinated induction of HAS2 and TSG-6 facilitation of pericellular HA coat assembly is necessary for TGF-beta1-dependent activation of fibroblasts, and both components of this response are impaired with in vitro aging. In conclusion, the HA pericellular coat is integral but not sufficient to correct for the age-dependent defect in phenotypic conversion.

Alteration in the glycosaminoglycan hyaluronan (HA) has been demonstrated in numerous renal diseases. We have demonstrated that renal proximal tubular epithelial cells (PTCs) surround themselves in vitro with HA in an organized pericellular matrix or 'coat', which is associated with cell migration, and also form pericellular HA cable-like structures which modulate PTC-mononuclear leukocytes interactions. The aim of this study was to characterize potential regulatory mechanism in the assembly of PTC-HA into pericellular cables. HA cables are generated by PTCs in the absence of serum. Immunohistochemical analysis demonstrates the incorporation of components of the inter-alpha-inhibitor (IalphaI) family of proteins and versican into HA cables. Addition of an antibody to IalphaI/PalphaI (pre-alpha-inhibitor) inhibits cable formation. In contrast, inhibition of tumor necrosis factor-alpha-stimulated gene 6 (TSG-6) has no effect on cable formation, suggesting that their generation is independent of the known heavy-chain transfer activity of TSG-6. Overexpression of HAS3 is associated with induction of HA cable formation, and also increased incorporation of HA into pericellular coats. Functionally, this resulted in enhanced HA-dependent monocyte binding and cell migration, respectively. Cell surface expression of CD44 and trypsin-released cell-associated HA were increased in HAS3-overexpressing cells. In addition, hyaluronidase (hyal1 and hyal2) and bikunin mRNA expression were increased, whereas PalphaI HC3 mRNA expression was unchanged in the transfected cells. The data demonstrate the importance of IalphaI/PalphaI in cable formation and suggest that expression of HAS3 may be critical for HA cable assembly.