bims-supasi Biomed News
on Sulfation pathways and signalling
Issue of 2025–03–09
twelve papers selected by
Jonathan Wolf Mueller, University of Birmingham
  1. Development of a method to measure the activity of heparan sulfate 6-endosulfatase for biological research.
  2. Glycosaminoglycan Mass Spectrometry Imaging by Infrared Matrix-Assisted Laser Desorption Electrospray Ionization.
  3. Heparan sulfate regulates amphiregulin programming of tissue reparative lung mesenchymal cells during influenza A virus infection in mice.
  4. Chondroitin sulfate binds to main protease of SARS-CoV-2 and efficaciously inhibits its activity.
  5. Unique structure and biological properties of fucosylated glycosaminoglycan and its oligosaccharides from sea cucumber Holothuria floridana.
  6. Synthetic heparan sulfate mimics based on chitosan derivatives show broad-spectrum antiviral activity.
  7. Intracytoplasmic accumulation of keratan sulfate is a hallmark of granular cell tumor.
  8. Injectable and drug-loaded gelatin methacrylate and carboxymethylated-sulfated xanthan gum hydrogels as biomimetic mineralization constructs.
  9. Caution, these are glycan sulfates. The features of their interaction with proteins.
  10. Characterizing the Sulfated and Glucuronidated (Poly)phenol Metabolome for Dietary Biomarker Discovery.
  11. Long-term exposure of indoxyl sulfate induces mesothelial-to-mesenchymal transition of peritoneal mesothelial cells via β-catenin-involved signaling pathway.
  12. Non-Anticoagulant Heparin: An In Vitro Investigation of a Novel Therapeutic Approach for Oral Cancer.
  1. Glycobiology. 2025 Mar 05. pii: cwaf012. [Epub ahead of print]
    Development of a method to measure the activity of heparan sulfate 6-endosulfatase for biological research.
    Zhangjie Wang, Julius Benicky, Pritha Mukherjee, Justin Laing, Yongmei Xu, Vijayakanth Pagadala, Shuangni Wu, Joseph A Hippensteel, Radoslav Goldman, Jian Liu.
      Heparan sulfate 6-endosulfatases (SULFs) remove 6-O-sulfo groups from heparan sulfate polysaccharide chains. SULFs modify the functions of heparan sulfate and contribute to the development of cancers, organ development and endothelial inflammatory responses. However, direct measurement of the activity of SULFs from human and mouse plasma is not currently possible. Here, we report a liquid chromatography coupled with tandem mass spectrometry (LS-MS/MS) assay to measure the activity of SULFs. The method uses a structurally homogeneous heparan sulfate dodecasaccharide (12-mer) in which the glucuronic and iduronic acid residues are labeled with both 13C- and 2H-atoms. The 12-mers desulfated by the SULFs is subjected to degradation with heparin lyases to yield disaccharides, which is followed by LC-MS/MS. The amount of two specific disaccharides, ΔIIIS and ΔIVS, quantified by LC-MS/MS reports the activity of the SULFs with high sensitivity and specificity. This method allows for the determination of the activity from conditioned cell media and mouse plasma. Our findings offer an essential novel tool to delineate many roles of SULFs in biological processes.
    Keywords:  LC–MS; endosulfatase; heparan sulfate; oligosaccharides
    DOI:  https://doi.org/10.1093/glycob/cwaf012
  2. J Am Soc Mass Spectrom. 2025 Mar 03.
    Glycosaminoglycan Mass Spectrometry Imaging by Infrared Matrix-Assisted Laser Desorption Electrospray Ionization.
    Tana V Palomino, David C Muddiman.
      Chondroitin sulfate (CS) is a type of glycosaminoglycan (GAG) that is abundant in cartilage and perineural networks (PNNs). Changes in the CS signature of PNNs have been implicated in several neurological diseases. Most CS-GAGs contain labile sulfate groups, which can be lost during ionization events that deposit large amounts of internal energy. Infrared matrix-assisted laser desorption electrospray ionization (IR-MALDESI) is a soft ionization technique used for mass spectrometry imaging. In this work, we determine the spatial distribution of CS-GAG disaccharides within rodent brain using IR-MALDESI MSI. Non-, mono-, and disulfated disaccharides were detected with various adducts. All disaccharides colocalized to the PNNs, which are most abundant in the cortex and hippocampus regions of the brain. This is the first MSI study to spatially resolve CS-GAG disaccharides within brain, paving the way for IR-MALDESI to measure GAGs in neurological diseases.
    Keywords:  Chondroitin Sulfate; Glycosaminoglycans; IR-MALDESI; Mass Spectrometry Imaging
    DOI:  https://doi.org/10.1021/jasms.4c00435
  3. Nat Commun. 2025 Mar 03. 16(1): 2129
    Heparan sulfate regulates amphiregulin programming of tissue reparative lung mesenchymal cells during influenza A virus infection in mice.
    Lucas F Loffredo, Anmol Kustagi, Olivia R Ringham, Fangda Li, Kenia de Los Santos-Alexis, Anjali Saqi, Nicholas Arpaia.
      Amphiregulin (Areg), a growth factor produced by regulatory T (Treg) cells to facilitate tissue repair, contains a heparan sulfate (HS) binding domain. How HS, a highly sulfated glycan subtype that alters growth factor signaling, influences Areg repair functions is unclear. Here we report that inhibition of HS in various cell lines and primary lung mesenchymal cells (LMC) qualitatively alters Areg downstream signaling. Utilization of a panel of cell lines with targeted deletions in HS synthesis-related genes identifies the glypican family of HS proteoglycans as critical for Areg signaling. In the context of influenza A virus (IAV) infection in vivo, an Areg-responsive subset of reparative LMC upregulate glypican-4 and HS; conditional deletion of HS primarily within this LMC subset results in reduced repair characteristics following IAV infection. This study demonstrates that HS on a specific lung mesenchymal population is a mediator of Treg cell-derived Areg reparative signaling.
    DOI:  https://doi.org/10.1038/s41467-025-57362-z
  4. Int J Biol Macromol. 2025 Feb 26. pii: S0141-8130(25)02098-7. [Epub ahead of print]306(Pt 2): 141547
    Chondroitin sulfate binds to main protease of SARS-CoV-2 and efficaciously inhibits its activity.
    Jinwen Li, Shu Jie Li.
      Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is constantly mutating and spreading globally, posing a great threat to people's lives and health. The main protease of SARS-CoV-2 (Mpro, also called 3CLpro) is an attractive drug target for SARS-CoV-2, due to its crucial role in processing the viral replication. Here, we showed that chondroitin sulfate (CS) from pig, cattle and shark efficaciously inhibits Mpro activity of SARS-CoV-2 with half maximal inhibitory concentrations (IC50) of 0.148, 0.121 and 0.119 μM, respectively, through a fluorescence resonance energy transfer (FRET) assay. The inhibition pattern of CSs against Mpro activity is competitive inhibition, with inhibition constants (Ki) of CSs derived from pig, bovine and shark are 0.111, 0.096, and 0.107 μM, respectively, indicating significant inhibitory effects of CSs on Mpro activity. Protein fluorescence quenching demonstrated that porcine, bovine and shark CSs strongly bind to Mpro protein with dissociation constants (KD) of 28.31, 28.47 and 20.66 μM at 25 °C at a physiological condition, respectively, mainly through van der Waals and hydrogen bond interactions. Molecular docking and dynamics analysis provides an insight into structural information of the binding of the CSs with Mpro protein. Our findings suggested that CSs from different origins might be a promising food ingredient for the prevention of the SARS-CoV-2 infection.
    Keywords:  Activity; Chondroitin sulfate; Inhibition; Interaction; Main protease; SARS-CoV-2
    DOI:  https://doi.org/10.1016/j.ijbiomac.2025.141547
  5. Carbohydr Polym. 2025 May 01. pii: S0144-8617(25)00132-8. [Epub ahead of print]355 123351
    Unique structure and biological properties of fucosylated glycosaminoglycan and its oligosaccharides from sea cucumber Holothuria floridana.
    Chen Xu, Xiang Shi, Huifang Sun, Lijuan Yu, Liang Zhang, Di Lan, Xiaolu Wu, Mengran Chen, Nanqi Cheng, Ying Pan, Jiayi He, Ronghua Yin, Lutan Zhou, Na Gao, Jinhua Zhao.
      Fucosylated glycosaminoglycan (FG) from Holothuroidea exhibits notable structural diversity and multiple biological activities. This study investigated the HfFG isolated from the sea cucumber Holothuria floridana, focusing on its chemical structure and biological activities. Structural analysis of eleven oligosaccharides (a-k) and a depolymerized product (dHfFG-II) using NMR identified the HfFG backbone as chondroitin sulfate E (CS-E), with various branches, including L-Fuc2S4S, L-Fuc3S4S, L-Fuc4S, and the unique disaccharide D-GalNAc4S-α1,2-L-Fuc3S4S, attached at C-3 of GlcA. A high L-Fuc2S4S (40 %) and disaccharide branch (35 %) content allowed their contiguous distribution within the CS-E chain, as evidenced by the predominant hexasaccharides (i, j) in size-homogeneous Fr4 and the novel nonasaccharide k in Fr5. Notably, previously unreported branches and sequences in HfFG were confirmed, offering new understanding of the natural HfFG structure. HfFG showed potent inhibitory activities on the intrinsic tenase complex (iXase), heparanase, and P-selectin binding to PSGL-1. Depolymerization selectively modulated these activities, preserving anti-iXase potency while attenuating heparanase and P-selectin inhibition. These activities were dependent on oligosaccharide chain length and sequence. Comparing the activities of FG and its oligosaccharides highlights their potential for the rational design of targeted inhibitors of heparanase or P-selectin binding to PSGL-1, with significant implications for therapeutic applications.
    Keywords:  Disaccharide branches; Fucosylated glycosaminoglycan; Heparanase; Oligosaccharides; P-selectin; iXase
    DOI:  https://doi.org/10.1016/j.carbpol.2025.123351
  6. Commun Biol. 2025 Mar 04. 8(1): 360
    Synthetic heparan sulfate mimics based on chitosan derivatives show broad-spectrum antiviral activity.
    IBV-Covid19-Pipeline
      Enveloped viruses enter cells by binding to receptors present on host cell membranes, which trigger internalization and membrane fusion. For many viruses, this either directly or indirectly involves interaction with membrane-anchored carbohydrates, such as heparan sulfate, providing a potential target for a broad-spectrum antiviral approach. Based on this hypothesis, we screened a library of functionalized chitosan sulfates that mimic heparan sulfate in cellular membranes for inhibition of SARS-CoV-2 and respiratory syncytial virus (RSV) entry. An array of compounds blocking SARS-CoV-2 and RSV were identified, with the lead compound displaying broad-spectrum activity against multiple viral strains and clinical isolates. Mechanism of action studies showed the drug to block viral entry irreversibly, likely via a virucidal mechanism. Importantly, the drug was non-toxic in vivo and showed potent post-exposure therapeutic activity against both SARS-CoV-2 and RSV. Together, these results highlight the potential of functionalized carbohydrates as broad-spectrum antivirals targeting respiratory viruses.
    DOI:  https://doi.org/10.1038/s42003-025-07763-z
  7. Pathol Res Pract. 2025 Mar 03. pii: S0344-0338(25)00084-6. [Epub ahead of print]269 155892
    Intracytoplasmic accumulation of keratan sulfate is a hallmark of granular cell tumor.
    Hitomi Hoshino, Akifumi Muramoto, Tomoya O Akama, Hisato Yoshida, Natsumi Yonemoto, Mana Fukushima, Motohiro Kobayashi.
      Granular cell tumor (GCT) is a relatively rare neoplasm characterized by abundant eosinophilic intracytoplasmic granules. More than three decades ago, Ehara and Katsuyama reported that GCT granules are recognized by the anti-keratan sulfate (KS) monoclonal antibody 5D4 and suggested that 5D4 could serve as a diagnostic marker for GCT. However, due to the small number of samples analyzed and incomplete structural analysis of KS, use of 5D4 as a GCT marker has not yet been widely accepted. To confirm its use as a GCT marker, we performed quantitative immunohistochemical analysis of GCT (n = 27) and other GCT-mimicking tumors/lesions (n = 82) including schwannoma (n = 10), neurofibroma (n = 10), melanocytic nevus (n = 10), leiomyoma (n = 10), gastrointestinal stromal tumor (GIST) (n = 10), malignant melanoma (n = 10), dermatofibroma (n = 10), angiosarcoma (n = 10) and malakoplakia (n = 2) using 5D4 and two other anti-KS monoclonal antibodies, R-10G and 297-11A, in combination with two endoglycosidases, keratanase II and endo-β-galactosidase. We found that most GCT tissues were immunostained with 5D4 and 297-11A, although tumors/lesions mimicking GCT showed minimal staining. Moreover, structural analysis revealed that KS accumulated in GCT consisted of both highly sulfated and low-sulfated KS located at non-reducing and reducing termini, respectively. Here we propose that intracytoplasmic KS accumulation is a hallmark of GCT, and that 5D4 and 297-11A could serve as diagnostic markers of GCT.
    Keywords:  297–11A; 5D4; Diagnostic marker; Granular cell tumor; Keratan sulfate
    DOI:  https://doi.org/10.1016/j.prp.2025.155892
  8. Carbohydr Polym. 2025 May 01. pii: S0144-8617(25)00135-3. [Epub ahead of print]355 123354
    Injectable and drug-loaded gelatin methacrylate and carboxymethylated-sulfated xanthan gum hydrogels as biomimetic mineralization constructs.
    Shiyao Liu, Fan Han, Pu Chen, Ruquan Zhang, Yongzhen Tao.
      Injectable and drug-loaded hydrogels based on gelatin and xanthan gum derivatives were biomineralized to form organic-inorganic hybrid composites with osteoconductivity and controllable release of antibiotic drug for inducing bone generation. Gelatin was amidated to get gelatin methacrylate (GM) for supporting cell adhesion and photo-crosslinkability. Xanthan gum was chemically modified to obtain carboxymethalated and sulfated derivatives (CMXG and SXG) with high negative charges for mimicking chondroitin sulfate in bone. GM was co-dissolved with CMXG/SXG and ciprofloxacin hydrochloride (CPFXH), and photo-crosslinked with lithium phenyl-2,4,6-tri methylbenzoylphosphinate (LAP) to fabricate drug-loaded CMXG/SXG-GM-CPFXH-LAP hydrogels, which possessed swelling ratio of 1.30 ± 0.03 and controlled release of CPFXH in PBS for 24 h. The 7d-mineralized CMXG/SXG3-GM12-CPFXH-LAP hydrogel showed dense mineral layers with Ca/P atomic ratio of 1.79, degree of crystalline of 77.3 %, mineral content of 50.8 %, and 2.6 times higher shear modulus than original one. The CMXG/SXG3-GM12-CPFXH-LAP solution was acted as "inks" to "write" word (BONE) and Chinese character ("Gu") manually, and was transferred into moulds to obtain hydrogel constructs with good fidelity of patterns, suggesting injectability and printability. The injectable, mineralizable, biocompatible and drug-loaded CMXG/SXG-GM-CPFXH-LAP hydrogels possess promising applications in bone tissue engineering due to facilitating osteoconductivity, recruiting cells, and reducing inflammation.
    Keywords:  Biomimetic mineralization; Carboxymethylated xanthan gum; Drug-loaded hydrogel; Gelatin methacrylate; Injectability; Sulfated xanthan gum
    DOI:  https://doi.org/10.1016/j.carbpol.2025.123354
  9. Carbohydr Res. 2025 Feb 20. pii: S0008-6215(25)00059-X. [Epub ahead of print]552 109433
    Caution, these are glycan sulfates. The features of their interaction with proteins.
    Nicolai Bovin, Marina Sablina, Galina Pazynina, Polina Obukhova, Aligeydar Ragimov, Nadezhda Shilova.
      In studies of viruses, lectins and especially human blood anti-glycan antibodies using printed glycan array (PGA), sulfated glycans suspiciously often turn out to be the highest-level binders. The binding to sulfated glycan along with parent neutral is easily explained by the similarity of these two glycans, while the unexpected thing is the many times stronger binding. Analysis of data accumulated over almost two decades allows us to explain the observed effect by the Coulomb interaction of the sulfate residue with a positively charged amino acid that accidently appears near the binding site of the neutral glycan backbone. That is, there is an effect of enhancing the specific interaction by an additional electrostatic one. It is expected that the material considered in the article will be useful for the correct interpretation of other data on the specificity of proteins capable of binding charged glycans, which are often encountered in nature.
    Keywords:  Anti-glycan antibodies; Electrostatic interaction; Immunoglobulins; Lectin; Printed glycan array; Sulfated glycans
    DOI:  https://doi.org/10.1016/j.carres.2025.109433
  10. J Agric Food Chem. 2025 Mar 02.
    Characterizing the Sulfated and Glucuronidated (Poly)phenol Metabolome for Dietary Biomarker Discovery.
    Ioanna Tsiara, Belén Hervás Povo, Wafa Alotaibi, Paul Young Tie Yang, Ana Rodriguez-Mateos, Daniel Globisch.
      (Poly)phenols, bioactive compounds abundant in plant-based diets, have attracted interest for their potential role in preventing chronic diseases including cardiometabolic and neurodegenerative diseases. This study investigates the global sulfatome and glucuronidated metabolome in urine samples from 100 healthy adults collected pre- and postintervention following a 3-day (poly)phenol-rich intervention consisting of flaxseeds, raspberry powder, and soy milk. Using untargeted mass spectrometric metabolomics combined with selective phase II enzymatic treatment, we detected 156 sulfated and 143 glucuronidated metabolites in urine samples. Significant changes postintervention were observed for 91 sulfates and 94 glucuronides. Receiver operating characteristic curve analysis identified a combination of six polyphenol-derived key metabolites: glucuronidated daidzein and the sulfated compounds of pyrogallol, ferulic acid, 4-methoxyphenol, enterolactone, and resorcinol, which resulted in the best combination with the highest predictive AUC of 0.97. These findings underscore the utility of these metabolites as sensitive and selective biomarkers of (poly)phenol dietary intake.
    Keywords:  (poly)phenols; gut microbiota; metabolomics; nutritional biomarkers; phase II modifications
    DOI:  https://doi.org/10.1021/acs.jafc.4c12596
  11. Adv Clin Exp Med. 2025 Mar 06.
    Long-term exposure of indoxyl sulfate induces mesothelial-to-mesenchymal transition of peritoneal mesothelial cells via β-catenin-involved signaling pathway.
    Runmei Liu, Wen Wen, Qiang Wang, Xiaoxue Weng, Guoqing Yu.
       BACKGROUND: Long-term peritoneal dialysis (PD) leads to peritoneal injury, with mesothelial-to-mesenchymal transition (MMT) potentially serving as an initial and reversible stage of this process. Indoxyl sulfate (IS), a protein-bound uremic toxin that accumulates in patients with declining renal function, is known to be associated with epithelial-mesenchymal transition (EMT) in proximal renal tubular cells. However, its effects on peritoneal mesothelial cells, which serve as the first-line barrier during PD, have not yet been investigated.
    OBJECTIVES: This study aimed to evaluate whether IS induces MMT in human peritoneal mesothelial cells during PD through the β-catenin signaling pathway.
    MATERIAL AND METHODS: A human peritoneal mesothelial cell line (HMrSV5) was used for this in vitro study. Cells were treated with IS or combined with β-catenin inhibitor ICG-001, and high glucose PD fluid (PDF) served as a positive control. Morphology, proliferation and adhesion were assessed, while the expression of β-catenin and α-smooth muscle actin (α-SMA) as mesenchymal markers, along with E-cadherin as a mesothelial marker, were analyzed at both RNA and protein levels using real-time polymerase chain reaction (PCR) and western blot, respectively.
    RESULTS: The number of viable and adherent cells was significantly increased in the IS and PDF groups compared to the control (p < 0.05). Treatment with ICG-001 significantly reduced both viable and adherent cell numbers compared to cells treated with IS or PDF alone (p < 0.05). At the RNA level, IS treatment significantly decreased E-cadherin expression (p = 0.002) while significantly increasing β-catenin (p = 0.001) and α-SMA (p = 0.002) expression compared to the control group. These changes were reversed by ICG-001 treatment. Protein expression showed similar trends.
    CONCLUSIONS: Indoxyl sulfate induces MMT in human peritoneal mesothelial cells, and these changes can be reversed by the specific β-catenin inhibitor ICG-001. This suggests that IS may be considered as another inducer of MMT during PD through the β-catenin signaling pathway.
    Keywords:  epithelial–mesenchymal transition; indoxyl sulfate; mesothelial-to-mesenchymal transition; peritoneal dialysis; β-catenin
    DOI:  https://doi.org/10.17219/acem/195869
  12. Oral Dis. 2025 Mar 03.
    Non-Anticoagulant Heparin: An In Vitro Investigation of a Novel Therapeutic Approach for Oral Cancer.
    S A Hamza, R Paolini, N M O'Brien-Simpson, W Singleton, R Patini, M McCullough, A Celentano.
       BACKGROUND: Oral squamous cell carcinoma (OSCC) represents a significant global oral health concern. Non-anticoagulant heparin (NH) emerges as a promising solution considering the enhanced survival observed with anticoagulants in cancer treatment.
    METHODS: We used the MTS assay (0/24/48/72 h), scratch assay (MuviCyte, 0-18 h), invasion Matrigel (24 h), and cytotoxic assay (0-24 h) to assess the in vitro effects of NH and heparin (10, 20, 40, 80 U/mL) on three oral human cell lines (H400/H357/OKF6) as well as their ability to interfere with the chemotherapeutic agents 5FU and cisplatin (1-5 μg/mL).
    RESULTS: Remarkably, NH not only significantly induced a significant cytotoxic effect on both cancer cell lines at 80 U/mL but also inhibited proliferation at 48/72 h to a comparable extent as heparin. Notably, neither drug exhibited cytotoxic effects on the normal cells. Furthermore, in H400/H357 cells, both heparin and NH significantly inhibit the cell migration and invasion rate. Importantly, the combination of these drugs with commonly used chemotherapeutic agents for OSCC treatment did not compromise their efficacy against the tested cell lines.
    CONCLUSION: NH demonstrates promising potential without compromising the efficacy of commonly used chemotherapeutic agents. These results underscore the need for the translation of this research to preclinical animal models.
    Keywords:  OSCC; chemotherapeutic agents; heparin; non‐anticoagulant heparin; oral cancer
    DOI:  https://doi.org/10.1111/odi.15288
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