bims-supasi Biomed News
on Sulfation pathways and signalling
Issue of 2023‒03‒12
fourteen papers selected by
Jonathan Wolf Mueller
University of Birmingham

  1. J Cell Sci. 2023 Mar 08. pii: jcs.260525. [Epub ahead of print]
      Morphogens provide quantitative and robust signaling systems to achieve stereotypic patterning and morphogenesis. Heparan sulfate proteoglycans (HSPGs) are key components of such regulatory feedback networks. In Drosophila, HSPGs serve as co-receptors for a number of morphogens, including Hedgehog (Hh), Wingless (Wg), Decapentaplegic (Dpp), and Unpaired (Upd). Recently, Windpipe (Wdp), a chondroitin sulfate proteoglycan (CSPG), was found to negatively regulate Upd and Hh signaling. However, roles of Wdp, and CSPGs in general, in morphogen signaling networks are poorly understood. We found that Wdp is a major CSPG with 4-O sulfated CS in Drosophila. Overexpression of wdp modulates Dpp and Wg signaling, showing that it is a general regulator of HS-dependent pathways. Although wdp mutant phenotypes are mild in the presence of morphogen signaling buffering systems, this mutant in the absence of Sulf1 or Dally, molecular hubs of the feedback networks, produces high levels of synthetic lethality and various severe morphological phenotypes. Our study indicates a close functional relationship between HS and CS, and identifies the Wdp CSPG as a novel component in morphogen feedback pathways.
    Keywords:   Drosophila ; Chondroitin sulfate proteoglycan; Decapentaplegic; Morphogen signaling; Windpipe; Wingless
  2. Carbohydr Polym. 2023 Jan 01. pii: S0144-8617(22)01081-5. [Epub ahead of print]299 120176
      Tau spreading in Alzheimer's disease is mediated by cell surface heparan sulfate (HS). As a class of sulfated polysaccharides, fucoidans might compete with HS to bind tau, resulting in the cessation of tau spreading. The structural determinants of fucoidans for competition with HS binding to tau are not well understood. Sixty previously prepared fucoidans/glycans with different structural determinants were used to determine their binding abilities to tau using SPR and AlphaLISA. Finally, it was found that fucoidans had two fractions (sulfated galactofucan (SJ-I) and sulfated heteropolysaccharide (SJ-GX-3)), which exhibited strong binding abilities than heparin. Tau cellular uptake assays using wild type mouse lung endothelial cell lines were performed. It was shown SJ-I and SJ-GX-3 inhibited tau-cell interaction and tau cellular uptake, suggesting that fucoidans might be good candidates for inhibiting tau spreading. NMR titration mapped fucoidans binding sites, which could provide the theoretical basis for the design of tau spreading inhibitors.
    Keywords:  Alzheimer's disease; Fucoidans; Heparin; Sulfated glycan; Tau protein
  3. Carbohydr Polym. 2023 Jan 01. pii: S0144-8617(22)01096-7. [Epub ahead of print]299 120191
      Glucuronyl 5-epimerase (Hsepi) converts D-glucuronic acid (GlcA) into L-iduronic acid (IdoA) units, through a mechanism involving reversible abstraction of a proton at C5 of hexuronic acid residues. Incubations of a [4GlcAβ1-4GlcNSO3α1-]n precursor substrate with recombinant enzymes in a D2O/H2O medium enabled an isotope exchange approach to the assessment of functional interactions of Hsepi with hexuronyl 2-O-sulfotransferase (Hs2st) and glucosaminyl 6-O-sulfotransferase (Hs6st), both involved in the final polymer-modification steps. Enzyme complexes were supported by computational modeling and homogeneous time resolved fluorescence. GlcA and IdoA D/H ratios related to product composition revealed kinetic isotope effects that were interpreted in terms of efficiency of the coupled epimerase and sulfotransferase reactions. Evidence for a functional Hsepi/Hs6st complex was provided by selective incorporation of D atoms into GlcA units adjacent to 6-O-sulfated glucosamine residues. The inability to achieve simultaneous 2-O- and 6-O-sulfation in vitro supported topologically separated reactions in the cell. These findings provide novel insight into the roles of enzyme interactions in heparan sulfate biosynthesis.
    Keywords:  2-O-sulfotransferase; 6-Osulfotransferase; Glucuronosyl C5-epimerase; Heparan sulfate
  4. Sci Data. 2023 Mar 06. 10(1): 121
      Heparan sulfates (HS) proteoglycans are commonly found on the cell surface and mediate many processes. Binding of HS ligands is determined by the sulfation code on the HS chain that can be N-/2-O/6-O- or 3-O-sulfated, generating heterogenous sulfation patterns. 3-O sulfated HS (3S-HS) play a role in several (patho)physiological processes such as blood coagulation, viral pathogenesis and binding and internalization of tau in Alzheimer's disease. However, few 3S-HS-specific interactors are known. Thus, our insight into the role of 3S-HS in health and disease is limited, especially in the central nervous system. Using human CSF, we determined the interactome of synthetic HS with defined sulfation patterns. Our affinity-enrichment mass spectrometry studies expand the repertoire of proteins that may interact with (3S-)HS. Validating our approach, ATIII, a known 3S-HS interactor, was found to require GlcA-GlcNS6S3S for binding, similar to what has been reported. Our dataset holds novel, potential HS and 3S-HS protein ligands, that can be explored in future studies focusing on molecular mechanisms that depend on 3S-HS in (patho)physiological conditions.
  5. Carbohydr Polym. 2023 Jan 01. pii: S0144-8617(22)01093-1. [Epub ahead of print]299 120188
      Most bone repair scaffolds are multi-connected channel structure, but the hollow structure is not conducive to the transmission of active factors, cells and so on. Here, microspheres were covalently integrated into 3D-printed frameworks to form composite scaffolds for bone repair. The frameworks composed of double bond modified gelatin (Gel-MA) and nano-hydroxyapatite (nHAP) provided strong support for related cells climbing and growth. Microspheres, which were made of Gel-MA and chondroitin sulfate A (CSA), were able to connect the frameworks like bridges, providing channels for cells migration. Additionally, CSA released from microspheres promoted the migration of osteoblasts and enhanced osteogenesis. The composite scaffolds could effectively repair mouse skull defect and improve MC3T3-E1 osteogenic differentiation. These observations confirm the bridging effect of microspheres rich in chondroitin sulfate and also determine that the composite scaffold can be as a promising candidate for enhanced bone repair.
    Keywords:  3D printing; Bone regeneration; Chondroitin sulfate; Composite scaffold; Microsphere
  6. Carbohydr Polym. 2023 Jan 01. pii: S0144-8617(22)01072-4. [Epub ahead of print]299 120167
      In December 2019, the global coronavirus disease 2019 (COVID-19) pandemic began in Wuhan, China. COVID-19 is caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), which infects host cells primarily through the angiotensin-converting enzyme 2 (ACE2) receptor. In addition to ACE2, several studies have shown the importance of heparan sulfate (HS) on the host cell surface as a co-receptor for SARS-CoV-2-binding. This insight has driven research into antiviral therapies, aimed at inhibiting the HS co-receptor-binding, e.g., by glycosaminoglycans (GAGs), a family of sulfated polysaccharides that includes HS. Several GAGs, such as heparin (a highly sulfated analog of HS), are used to treat various health indications, including COVID-19. This review is focused on current research on the involvement of HS in SARS-CoV-2 infection, implications of viral mutations, as well as the use of GAGs and other sulfated polysaccharides as antiviral agents.
    Keywords:  Glycosaminoglycans; Heparan sulfate co-receptor; Receptor binding domain; SARS-CoV-2; Spike protein
  7. Am J Physiol Cell Physiol. 2023 Mar 06.
      Alzheimer's disease (AD) is the most common form of dementia. Currently, there is no effective treatment for AD, as its etiology remains poorly understood. Mounting evidence suggests that the accumulation and aggregation of amyloid-β peptides (Aβ), which constitute amyloid plaques in the brain, is critical for initiating and accelerating AD pathogenesis. Considerable efforts have been dedicated to shedding light on the molecular basis and fundamental origins of the impaired Aβ metabolism in AD. Heparan sulfate (HS), a linear polysaccharide of the glycosaminoglycan family, co-deposits with Aβ in plaques in the AD brain, directly binds and accelerates Aβ aggregation, and mediates Aβ internalization and cytotoxicity. Mouse model studies demonstrate that HS regulates Aβ clearance and neuroinflammation in vivo. Previous reviews have extensively explored these discoveries. Here, this review focuses on the recent advancements in understanding abnormal HS expression in the AD brain, the structural aspects of HS-Aβ interaction, and the molecules involved in modulating Aβ metabolism through HS interaction. Furthermore, this review presents a perspective on the potential effects of abnormal HS expression on Aβ metabolism and AD pathogenesis. Additionally, the review highlights the importance of conducting further research to differentiate the spatiotemporal components of HS structure and function in the brain and AD pathogenesis.
    Keywords:  Alzheimer's disease; amyloid-β; expression; heparan sulfate; metabolism
  8. Anal Methods. 2023 Mar 06.
      The fine structure of heparan sulfate (HS), the glycosaminoglycan polysaccharide component of cell surface and extracellular matrix HS proteoglycans, coordinates the complex cell signalling processes that control homeostasis and drive development in multicellular animals. In addition, HS is involved in the infection of mammals by viruses, bacteria and parasites. The current detection limit for fluorescently labelled HS disaccharides (low femtomole; 10-15 mol), has effectively hampered investigations of HS composition in small, functionally-relevant populations of cells and tissues that may illuminate the structural requirements for infection and other biochemical processes. Here, an ultra-high sensitivity method is described that utilises a combination of reverse-phase HPLC, with tetraoctylammonium bromide (TOAB) as the ion-pairing reagent and laser-induced fluorescence detection of BODIPY-FL-labelled disaccharides. The method provides an unparalleled increase in the sensitivity of detection by ∼six orders of magnitude, enabling detection in the zeptomolar range (∼10-21 moles; <1000 labelled molecules). This facilitates determination of HS disaccharide compositional analysis from minute samples of selected tissues, as demonstrated by analysis of HS isolated from the midguts of Anopheles gambiae mosquitoes that was achieved without approaching the limit of detection.
  9. Int J Biol Macromol. 2023 Mar 08. pii: S0141-8130(23)00846-2. [Epub ahead of print]236 123952
      Improving chronic wound healing remains a challenge in the clinical practice. In this study, we developed double-crosslinked angiogenic 3D-bioprinted patches for diabetic wound healing by the photocovalent crosslinking of vascular endothelial growth factor (VEGF) using ultraviolet (UV) irradiation. 3D printing technology can precisely customize the structure and composition of patches to meet different clinical requirements. The biological polysaccharide alginate and chondroitin sulfate methacryloyl were used as biomaterials to construct the biological patch, which could be crosslinked using calcium ion crosslinking and photocrosslinking, thereby improving its mechanical properties. More importantly, acrylylated VEGF could be easily and rapidly photocrosslinked under UV irradiation, which simplified the step of chemically coupling growth factors and prolonged VEGF release time. These characteristics suggest that 3D-bioprinted double-crosslinked angiogenic patches are ideal candidates for diabetic wound healing and other tissue engineering applications.
    Keywords:  3D-bioprinting; Alginate; Chondroitin sulfate; Diabetic wound healing; Patches
  10. Cells. 2023 Feb 25. pii: 744. [Epub ahead of print]12(5):
      Our previous studies demonstrated that enzymatic removal of highly sulfated heparan sulfates with heparinase 1 impaired axonal excitability and reduced expression of ankyrin G at the axon initial segments in the CA1 region of the hippocampus ex vivo, impaired context discrimination in vivo, and increased Ca2+/calmodulin-dependent protein kinase II (CaMKII) activity in vitro. Here, we show that in vivo delivery of heparinase 1 in the CA1 region of the hippocampus elevated autophosphorylation of CaMKII 24 h after injection in mice. Patch clamp recording in CA1 neurons revealed no significant heparinase effects on the amplitude or frequency of miniature excitatory and inhibitory postsynaptic currents, while the threshold for action potential generation was increased and fewer spikes were generated in response to current injection. Delivery of heparinase on the next day after contextual fear conditioning induced context overgeneralization 24 h after injection. Co-administration of heparinase with the CaMKII inhibitor (autocamtide-2-related inhibitory peptide) rescued neuronal excitability and expression of ankyrin G at the axon initial segment. It also restored context discrimination, suggesting the key role of CaMKII in neuronal signaling downstream of heparan sulfate proteoglycans and highlighting a link between impaired CA1 pyramidal cell excitability and context generalization during recall of contextual memories.
    Keywords:  ankyrin G; axon initial segment; axonal excitability; context discrimination; extracellular matrix; synaptic plasticity
  11. J Clin Med. 2023 Feb 28. pii: 1924. [Epub ahead of print]12(5):
      Non-suicidal self-injury (NSSI) is a significant public health issue that particularly affects female adolescents usually emerging during puberty, with a subsequent reduction and even remission in the phenomenon later in life. The dysregulation of the hormonal stress response, particularly cortisol and dehydroepiandrosterone sulfate (DHEA-S), whose levels increase markedly during pubertal adrenarche, has been associated with the development and maintenance of a wide range of emotional disorders. Our study aims to investigate whether different cortisol-DHEA-S response patterns could be associated with the main motivational moderators to engage NSSI as well as with urgency and motivation to stop NSSI in a sample of female adolescents. We found significant correlations between stress hormones and several factors that support and sustain NSSI, specifically: cortisol levels and distressing/upsetting urge (r = 0.39 and a p = 8.94 × 10-3) and sensation seeking (r = -0.32 and a p = 0.04), as well as cortisol/DHEA-s ratio and external emotion regulation (r = 0.40 and a p = 0.01) and desire to stop NSSI (r = 0.40 and a p = 0.01). Cortisol and DHEA-S may play a role in NSSI through the regulation of stress responses and affective states. Such results could have implications for the development of new and improved treatment and prevention plans for NSSI.
    Keywords:  DHEA-S; Ottawa Self Injury Inventory; cortisol; emotional dysregulation; non-suicidal self-injury; stress response; theory of urgency
  12. Cancers (Basel). 2023 Feb 28. pii: 1524. [Epub ahead of print]15(5):
      Historically, the only focus on tissue factor (TF) in clinical pathophysiology has been on its function as the initiation of the extrinsic coagulation cascade. This obsolete vessel-wall TF dogma is now being challenged by the findings that TF circulates throughout the body as a soluble form, a cell-associated protein, and a binding microparticle. Furthermore, it has been observed that TF is expressed by various cell types, including T-lymphocytes and platelets, and that certain pathological situations, such as chronic and acute inflammatory states, and cancer, may increase its expression and activity. Transmembrane G protein-coupled protease-activated receptors can be proteolytically cleaved by the TF:FVIIa complex that develops when TF binds to Factor VII (PARs). The TF:FVIIa complex can activate integrins, receptor tyrosine kinases (RTKs), and PARs in addition to PARs. Cancer cells use these signaling pathways to promote cell division, angiogenesis, metastasis, and the maintenance of cancer stem-like cells. Proteoglycans play a crucial role in the biochemical and mechanical properties of the cellular extracellular matrix, where they control cellular behavior via interacting with transmembrane receptors. For TFPI.fXa complexes, heparan sulfate proteoglycans (HSPGs) may serve as the primary receptor for uptake and degradation. The regulation of TF expression, TF signaling mechanisms, their pathogenic effects, and their therapeutic targeting in cancer are all covered in detail here.
    Keywords:  cancer; platelets; proteoglycans; syndecans; tissue factor
  13. Int J Biol Macromol. 2023 Mar 08. pii: S0141-8130(23)00842-5. [Epub ahead of print] 123948
      A sulfated polysaccharide from the brown alga Ishige okamurae Yendo, designated IOY, was successfully isolated by anion-exchange and size-exclusion chromatography. Chemical and spectroscopic analyses demonstrated that IOY was a fucoidan, that consisted of →3)-α-l-Fucp-(1→, →4)-α-l-Fucp-(1→, →6)-β-d-Galp-(1 → and →3)-β-d-Galp-(1 → residues with sulfate groups at C-2/C-4 the of (1 → 3)-α-l-Fucp and C-6 the of (1 → 3)-β-d-Galp residues. IOY possessed a potent immunomodulatory effect in vitro as measured by lymphocyte proliferation assay. The immunomodulatory effect of IOY was further investigated in vivo using immunosuppressed mice induced by cyclophosphamide (CTX). The results showed that IOY significantly increased the spleen and thymus indexes and alleviated CTX-induced spleen and thymus damage. Furthermore, IOY had a significant effect on hematopoietic function recovery and promoted the secretion of interleukin-2 (IL-2) and tumor necrosis factor (TNF-α). Notably, IOY reversed CD4+ and CD8+ T cell reduction and improved immune response. These data indicated that IOY had vital in immunomodulatory function and could be used as drug or functional food to lessen chemotherapy-induced immunosuppression.
    Keywords:  Fucoidan; Immunomodulatory activity; Ishige okamurae; Structural characterization
  14. Front Cardiovasc Med. 2023 ;10 990422
      Background: Patients with chronic kidney disease (CKD) have a high risk of left ventricular hypertrophy (LVH). Fibroblast growth factor 23 (FGF23) and indoxyl sulfate (IS) are associated with LVH in patients with CKD, but the interactions between these molecules remain unknown. We investigated whether IS contributes to LVH associated with FGF23 in cultured cardiomyocytes and CKD mice.Methods and results: In cultured rat cardiac myoblast H9c2 cells incubated with IS, mRNA levels of the LVH markers atrial natriuretic factor, brain natriuretic peptide, and β-myosin heavy chain were significantly upregulated. Levels of mRNA of the polypeptide N-acetylgalactosaminyltransferase 3 (GALNT3), which regulates FGF23 O-glycosylation, and FGF23 were also upregulated in H9c2 cells. Intact FGF23 protein expression and fibroblast growth factor receptor 4 (FGFR4) phosphorylation were increased in cell lysates by IS administration. In C57BL/6J mice with heminephrectomy, IS promoted LVH, whereas the inhibition of FGFR4 significantly reduced heart weight and left ventricular wall thickness in IS-treated groups. While there was no significant difference in serum FGF23 concentrations, cardiac FGF23 protein expression was markedly increased in IS-injected mice. GALNT3, hypoxia-inducible factor 1 alpha, and FGF23 protein expression was induced in H9c2 cells by IS treatment and suppressed by the inhibition of Aryl hydrocarbon receptor which is the receptor for IS.
    Conclusion: This study suggests that IS increases FGF23 protein expression via an increase in GALNT3 and hypoxia-inducible factor 1 alpha expression, and activates FGF23-FGFR4 signaling in cardiomyocytes, leading to LVH.
    Keywords:  aryl hydrocarbon receptor; fibroblast growth factor 23; fibroblast growth factor receptor 4; indoxyl sulfate; left ventricular hypertrophy