Human xylosyltransferase-I – A new marker for myofibroblast differentiation in skin fibrosis

Faust, Isabel; Roch, Christina; Kühn, Joachim; Prante, Christian; Knabbe, Cornelius; Hendig, Doris

doi:10.1016/j.bbrc.2013.05.125

Cited by 35 publications

(56 citation statements)

References 31 publications

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“…Biological samples were prepared in triplicates using passages 7–10. Cells harvested for RNA isolation and protein lysis were treated as described before [37,38]. …”

Section: Methodsmentioning

confidence: 99%

“…PCR array was performed with cells cultivated for 24 h without FCS, and 3 biological replicates were pooled for array analysis. RNA was isolated as described before [37], whereupon 400 ng were used for cDNA-synthesis (RT 2 First Strand Kit; Qiagen, Hilden, Germany). Real-Time PCR was performed using LightCycler® 480 (Roche, Penzberg, Germany) and RT 2 SYBR Green qPCR reaction mixture (Qiagen, Hilden, Germany).…”

Section: Methodsmentioning

confidence: 99%

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ABCC6- a new player in cellular cholesterol and lipoprotein metabolism?

et al. 2014

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BackgroundDysregulations in cholesterol and lipid metabolism have been linked to human diseases like hypercholesterolemia, atherosclerosis or the metabolic syndrome. Many ABC transporters are involved in trafficking of metabolites derived from these pathways. Pseudoxanthoma elasticum (PXE), an autosomal-recessive disease caused by ABCC6 mutations, is characterized by atherogenesis and soft tissue calcification.MethodsIn this study we investigated the regulation of cholesterol biosynthesis in human dermal fibroblasts from PXE patients and healthy controls.ResultsGene expression analysis of 84 targets indicated dysregulations in cholesterol metabolism in PXE fibroblasts. Transcript levels of ABCC6 were strongly increased in lipoprotein-deficient serum (LPDS) and under serum starvation in healthy controls. For the first time, increased HMG CoA reductase activities were found in PXE fibroblasts. We further observed strongly elevated transcript and protein levels for the proprotein convertase subtilisin/kexin type 9 (PCSK9), as well as a significant reduction in APOE mRNA expression in PXE.ConclusionIncreased cholesterol biosynthesis, elevated PCSK9 levels and reduced APOE mRNA expression newly found in PXE fibroblasts could enforce atherogenesis and cardiovascular risk in PXE patients. Moreover, the increase in ABCC6 expression accompanied by the induction of cholesterol biosynthesis supposes a functional role for ABCC6 in human lipoprotein and cholesterol homeostasis.

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“…Biological samples were prepared in triplicates using passages 7–10. Cells harvested for RNA isolation and protein lysis were treated as described before [37,38]. …”

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

ABCC6- a new player in cellular cholesterol and lipoprotein metabolism?

et al. 2014

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“…To investigate the effect of the cytokines and inhibitors, NHDF were cultured for 24 h before treatment using a formerly established cell culture model [26]. In brief, cells were cultivated with a low density of 50 cells per mm 2 on a hard tissue culture substrate (100 mm × 20 mm dish), promoting their trans-differentiation into proto-myofibroblasts.…”

Section: Cytokine and Inhibitor Treatmentmentioning

confidence: 99%

“…Previous work has demonstrated that TGFβ1 is a potent inducer of the XT-I gene and protein expression in primary human dermal and cardiac fibroblasts [26,27]. We wanted to examine whether other pro-fibrotic cytokines and growth factors are able to modulate XYLT1 and XYLT2 mRNA expression of skin fibroblasts.…”

Section: Identification Of Pro-fibrotic Mediators Regulating Xt-i Expmentioning

confidence: 99%

Activin A-Mediated Regulation of XT-I in Human Skin Fibroblasts

Plümers

Fischer

et al. 2020

Biomolecules

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Fibrosis is a fundamental feature of systemic sclerosis (SSc) and is characterized by excessive accumulation of extracellular matrix components like proteoglycans (PG) or collagens in skin and internal organs. Serum analysis from SSc patients showed an increase in the enzyme activity of xylosyltransferase (XT), the initial enzyme in PG biosynthesis. There are two distinct XT isoforms—XT-I and XT-II—in humans, but until now only XT-I is associated with fibrotic remodelling for an unknown reason. The aim of this study was to identify new XT mediators and clarify the underlying mechanisms, in view of developing putative therapeutic anti-fibrotic interventions in the future. Therefore, we used different cytokines and growth factors, small molecule inhibitors as well as small interfering RNAs, and assessed the cellular XT activity and XYLT1 expression in primary human dermal fibroblasts by radiochemical activity assays and qRT-PCR. We identified a new function of activin A as a regulator of XYLT1 mRNA expression and XT activity. While the activin A-induced XT-I increase was found to be mediated by activin A receptor type 1B, MAPK and Smad pathways, the activin A treatment did not alter the XYLT2 expression. Furthermore, we observed a reciprocal regulation of XYLT1 and XYLT2 transcription after inhibition of the activin A pathway components. These results improve the understanding of the differential expression regulation of XYLT isoforms under pathological fibroproliferative conditions.

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“…Particularly, NP is abundant in large aggregating proteoglycans, such as aggrecan and versican, that provide the osmotic properties of the disk critical to withstanding the high compressive loads applied to the spine. 8,13,14 In chondrocytes, XT-1 expression is suppressed by IL-1b, with Sp1 and Sp3 transcription factors exerting opposing effects on transcription. GAGs are of critical importance in the regulation of NP cell function and organ development.…”

mentioning

confidence: 99%

Xylosyltransferase-1 Expression Is Refractory to Inhibition by the Inflammatory Cytokines Tumor Necrosis Factor α and IL-1β in Nucleus Pulposus Cells

Zhou

Markova

et al. 2015

The American Journal of Pathology

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We investigated whether expression of xylosyltransferase-1 (XT-1), a key enzyme in glycosaminoglycan biosynthesis, is responsive to disk degeneration and to inhibition by the inflammatory cytokines tumor necrosis factor α and IL-1β in nucleus pulposus (NP) cells. Analysis of human NP tissues showed that XT-1 expression is unaffected by degeneration severity; XT-1 and Jun, Fos, and Sp1 mRNA were positively correlated. Cytokines failed to inhibit XT-1 promoter activity and expression. However, cytokines decreased activity of XT-1 promoters containing deletion and mutation of the -730/-723 bp AP-1 motif, prompting us to investigate the role of AP-1 and Sp1/Sp3 in the regulation of XT-1 in healthy NP cells. Overexpression and suppression of AP-1 modulated XT-1 promoter activity. Likewise, treatment with the Sp1 inhibitors WP631 and mithramycin A or cotransfection with the plasmid DN-Sp1 decreased XT-1 promoter activity. Inhibitors of AP-1 and Sp1 and stable knockdown of Sp1 and Sp3 resulted in decreased XT-1 expression in NP cells. Genomic chromatin immunoprecipitation analysis showed AP-1 binding to motifs located at -730/-723 bp and -684/-677 bp and Sp1 binding to -227/-217 bp and -124/-114 bp in XT-1 promoter. These results suggest that XT-1 expression is refractory to the disease process and to inhibition by inflammatory cytokines and that signaling through AP-1, Sp1, and Sp3 is important in the maintenance of XT-1 levels in NP cells.

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Human xylosyltransferase-I – A new marker for myofibroblast differentiation in skin fibrosis

Cited by 35 publications

References 31 publications

ABCC6- a new player in cellular cholesterol and lipoprotein metabolism?

ABCC6- a new player in cellular cholesterol and lipoprotein metabolism?

Activin A-Mediated Regulation of XT-I in Human Skin Fibroblasts

Xylosyltransferase-1 Expression Is Refractory to Inhibition by the Inflammatory Cytokines Tumor Necrosis Factor α and IL-1β in Nucleus Pulposus Cells

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