Lowered Expression of Heparan Sulfate/Heparin Biosynthesis Enzyme N-Deacetylase/N-Sulfotransferase 1 Results in Increased Sulfation of Mast Cell Heparin

Dagälv, Anders; Holmborn, Katarina; Kjellén, Lena; Åbrink, Magnus

doi:10.1074/jbc.m111.303891

Cited by 30 publications

(17 citation statements)

References 47 publications

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“…6B ) may be protective 50 . Selective changes in disaccharide content may also be specific to individual cells as mast cell responses have demonstrated altered HS content with Ndst1 deficiency 51 . Studies reported by other groups do support a pro-inflammatory function for some GAGs.…”

Section: Discussionmentioning

confidence: 99%

Selective Deletion of Heparan Sulfotransferase Enzyme, Ndst1, in Donor Endothelial and Myeloid Precursor Cells Significantly Decreases Acute Allograft Rejection

Chen

Ambadapadi

Wakefield

et al. 2018

Sci Rep

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Early damage to transplanted organs initiates excess inflammation that can cause ongoing injury, a leading cause for late graft loss. The endothelial glycocalyx modulates immune reactions and chemokine-mediated haptotaxis, potentially driving graft loss. In prior work, conditional deficiency of the glycocalyx-modifying enzyme N-deacetylase-N-sulfotransferase-1 (Ndst1f/f TekCre+) reduced aortic allograft inflammation. Here we investigated modification of heparan sulfate (HS) and chemokine interactions in whole-organ renal allografts. Conditional donor allograft Ndst1 deficiency (Ndst1−/−; C57Bl/6 background) was compared to systemic treatment with M-T7, a broad-spectrum chemokine-glycosaminoglycan (GAG) inhibitor. Early rejection was significantly reduced in Ndst1−/− kidneys engrafted into wildtype BALB/c mice (Ndst1+/+) and comparable to M-T7 treatment in C57Bl/6 allografts (P < 0.0081). M-T7 lost activity in Ndst1−/− allografts, while M-T7 point mutants with modified GAG-chemokine binding displayed a range of anti-rejection activity. CD3+ T cells (P < 0.0001), HS (P < 0.005) and CXC chemokine staining (P < 0.012), gene expression in NFκB and JAK/STAT pathways, and HS and CS disaccharide content were significantly altered with reduced rejection. Transplant of donor allografts with conditional Ndst1 deficiency exhibit significantly reduced acute rejection, comparable to systemic chemokine-GAG inhibition. Modified disaccharides in engrafted organs correlate with reduced rejection. Altered disaccharides in engrafted organs provide markers for rejection with potential to guide new therapeutic approaches in allograft rejection.

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Section: Discussionmentioning

confidence: 99%

Selective Deletion of Heparan Sulfotransferase Enzyme, Ndst1, in Donor Endothelial and Myeloid Precursor Cells Significantly Decreases Acute Allograft Rejection

Chen

Ambadapadi

Wakefield

et al. 2018

Sci Rep

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“…Heparin and chondroitin sulfate analysis by RPIP-HPLC was performed as described previously [26] with 5 x 10 6 BMMCs cultured either under normal copper conditions, excess copper (20 μM) or were deprived of copper (BCS: 200 μM).…”

Section: Methodsmentioning

confidence: 99%

Copper Regulates Maturation and Expression of an MITF:Tryptase Axis in Mast Cells

Frisk

Kjellén

Kaler

et al. 2017

The Journal of Immunology

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Copper has previously been implicated in the regulation of immune responses, but the impact of this metal on mast cells is poorly understood. Here, we address this issue and show that copper starvation of mast cells causes increased granule maturation as indicated by higher proteoglycan content, stronger metachromatic staining and altered ultrastructure in comparison with non-treated cells, whereas copper overload has opposite effects. In contrast, copper status did not impact storage of histamine in mast cells, nor did alterations in copper levels affect the ability of mast cells to degranulate in response to IgE receptor crosslinking. A striking finding was decreased tryptase content in mast cells with copper overload, whereas copper starvation increased tryptase content. These effects were associated with corresponding shifts in tryptase mRNA levels, suggesting that copper affects tryptase gene regulation. Mechanistically, we found that alterations in copper status affected the expression of Mitf, a transcription factor critical for driving tryptase expression. We also found evidence supporting the concept that the effects on Mitf are dependent on copper-mediated modulation of MAPK signaling. Finally, we show that in MEDNIK syndrome, a condition associated with low copper levels and a hyper-allergenic skin phenotype including pruritis and dermatitis, the number of tryptase-positive mast cells is increased. Taken together, our findings reveal a hitherto unrecognized role for copper in regulation of mast cell gene expression and maturation.

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“…To this end we used an approach where the glycosaminoglycan (GAG) side chains of the proteoglycans were depolymerized to disaccharide units by enzymatic digestion, followed by HPLC separation to determine the structure of the respective disaccharides (26). This structural analysis revealed that chondroitin sulfate (composed of repeating disaccharide units of the general structure: hexuronic acid (HexA)-N-acetylgalactosamine (GalNAc)) was the main type of GAG produced by both WT and Ctr2 −/− BMMCs.…”

Section: Resultsmentioning

confidence: 99%

“…Heparin and chondroitin sulfate analysis by RPIP-HPLC was performed with 5 × 10 6 cells as described previously (26). …”

Section: Methodsmentioning

confidence: 99%

Ctr2 Regulates Mast Cell Maturation by Affecting the Storage and Expression of Tryptase and Proteoglycans

Öhrvik

Logeman

Noguchi

et al. 2015

The Journal of Immunology

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Copper (Cu) is essential for multiple cellular functions. Cellular uptake of Cu+ is carried out by the Ctr1 high affinity Cu transporter. The mobilization of endosomal Cu pools is regulated by a protein structurally similar to Ctr1, called Ctr2. It was recently shown that ablation of Ctr2 caused an increase in the concentration of Cu localized to endolysosomes. However, the biological significance of excess endolysosomal Cu accumulation has not been assessed. Here we addressed this issue by investigating the impact of Ctr2 deficiency on mast cells, a cell type unusually rich in endolysosomal organelles (secretory granules). We show that Ctr2−/− mast cells have increased intracellular Cu concentrations and that the absence of Ctr2 results in increased metachromatic staining, the latter indicating an impact of Ctr2 on the storage of proteoglycans in the secretory granules. In agreement with this, the absence of Ctr2 caused a skewed ratio between proteoglycans of heparin and chondroitin sulfate type, with increased amounts of heparin accompanied by a reduction of chondroitin sulfate. Moreover, transmission electron microscopy analysis revealed a higher number of electron dense granules in Ctr2−/− mast cells than in wild-type cells. The increase in granular staining and heparin content is compatible with an impact of Ctr2 on mast cell maturation and, in support of this, the absence of Ctr2 resulted in markedly increased mRNA expression, storage and enzymatic activity of tryptase. Taken together, the present study introduces Ctr2 and Cu as novel actors in the regulation of mast cell maturation and granule homeostasis.

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Lowered Expression of Heparan Sulfate/Heparin Biosynthesis Enzyme N-Deacetylase/N-Sulfotransferase 1 Results in Increased Sulfation of Mast Cell Heparin

Cited by 30 publications

References 47 publications

Selective Deletion of Heparan Sulfotransferase Enzyme, Ndst1, in Donor Endothelial and Myeloid Precursor Cells Significantly Decreases Acute Allograft Rejection

Selective Deletion of Heparan Sulfotransferase Enzyme, Ndst1, in Donor Endothelial and Myeloid Precursor Cells Significantly Decreases Acute Allograft Rejection

Copper Regulates Maturation and Expression of an MITF:Tryptase Axis in Mast Cells

Ctr2 Regulates Mast Cell Maturation by Affecting the Storage and Expression of Tryptase and Proteoglycans

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