bims-supasi 2024-08-25 papers

bims-supasi

Biomed News

on Sulfation pathways and signalling

Issue of 2024–08–25
eleven papers selected by
Jonathan Wolf Mueller, University of Birmingham

Introducing Ion Mobility Mass Spectrometry in Brain Glycosaminoglycomics: Application to Chondroitin/Dermatan Sulfate Octasaccharide Domains.
TREM2 on microglia cell surface binds to and forms functional binary complexes with heparan sulfate modified with 6-O-sulfation and iduronic acid.
Delivery of Nitric Oxide by Chondroitin Sulfate C Increases the Rate of Wound Healing through Immune Modulation.
In situ formed aldehyde-modified hyaluronic acid hydrogel with polyelectrolyte complexes of aldehyde-modified chondroitin sulfate and gelatin: An approach for minocycline delivery.
Analysis of indoxyl sulfate in biological fluids with emphasis on sample preparation techniques: A comprehensive analytical review.
Enhanced osteogenesis and antibacterial activity of dual-functional PEEK implants via biomimetic polydopamine modification with chondroitin sulfate and levofloxacin.
Prognostic significance of HS2ST1 expression in patients with hepatocellular carcinoma.
Sialylated keratan sulfates on MUC5B are Siglec-8 ligands in the human esophagus.
Substantial benefits of an inert biphenyl column for the analysis of steroids and their phase II metabolites in biological samples.
Hexavalent chromium uptake in rice (Oryza sativa L.) mediated by sulfate and phosphate transporters OsSultr1;2 and OsPht1;1.
Electrospun EVOH/AST-120 hybrid nanofiber membranes for removal of indoxyl sulfate from blood.

J Am Soc Mass Spectrom. 2024 Aug 23.

Introducing Ion Mobility Mass Spectrometry in Brain Glycosaminoglycomics: Application to Chondroitin/Dermatan Sulfate Octasaccharide Domains.

Mirela Sarbu, Daniela G Seidler, David E Clemmer, Alina D Zamfir.

  Glycosaminoglycans (GAGs) are sulfated linear O-glycan chains abundantly expressed in the extracellular matrix (ECM). Among GAGs, chondroitin sulfate (CS) and dermatan sulfate (DS) play important roles at the brain level, where the distribution and location of the sulfates within the CS/DS chains are responsible for numerous biological events. The diversity of the neural hybrid CS/DS expressed in the brain and the need to elucidate their structure gave rise to considerable efforts toward the development of analytical methods able to discover novel regularly and irregularly sulfated domains. In this context, we report here the introduction of ion mobility separation (IMS) mass spectrometry (MS) in brain glycosaminoglycomics. Based on IMS MS and tandem MS (MS/MS) by collision-induced dissociation (CID), we have developed a powerful approach for the screening and structural analysis of neural CS/DS and optimized and validated the method for the structural analysis of octasaccharides that were released from brain proteoglycans by β-elimination and pooled after chain depolymerization using chondroitin AC lyase. The IMS MS data revealed the separation of CS/DS octamers into mobility families based on the amount of sulfation. Among the discovered oversulfated domains, of major biological importance is the pentasulfated-[4,5-Δ-GlcAGalNAc(IdoAGalNAc)3], for which the (-) nanoESI IMS CID MS/MS analysis disclosed through the presence of distinct drift times, the incidence of two isomers. Moreover, the generated fragment ions revealed an uncommon trisulfated motif and an uncommon pentasulfated motif. Hence, using IMS MS and CID MS/MS, novel brain-related CS/DS domains of atypical sulfation patterns were discovered and structurally characterized in detail.

Keywords:  brain chondroitin/dermatan sulfate; ion mobility mass spectrometry; irregular sulfation pattern; structural analysis

DOI:  https://doi.org/10.1021/jasms.4c00159
J Biol Chem. 2024 Aug 17. pii: S0021-9258(24)02192-6. [Epub ahead of print] 107691

TREM2 on microglia cell surface binds to and forms functional binary complexes with heparan sulfate modified with 6-O-sulfation and iduronic acid.

Ilayda Ozsan McMillan, Li Liang, Guowei Su, Xuehong Song, Kelly Drago, Hua Yang, Claudia Alvarez, Amika Sood, James Gibson, Robert J Woods, Chunyu Wang, Jian Liu, Fuming Zhang, Tom J Brett, Lianchun Wang.

  The Triggering Receptor Expressed on Myeloid Cells-2 (TREM2), a pivotal innate immune receptor, orchestrates functions such as inflammatory responses, phagocytosis, cell survival, and neuroprotection. TREM2 variants R47H and R62H have been associated with Alzheimer's disease, yet the underlying mechanisms remain elusive. Our previous research established that TREM2 binds to heparan sulfate (HS) and variants R47H and R62H exhibit reduced affinity for HS. Building upon this groundwork, our current study delves into the interplay between TREM2 and HS and its impact on microglial function. We confirm TREM2's binding to cell surface HS and demonstrate that TREM2 interacts with HS, forming HS-TREM2 binary complexes on microglia cell surfaces. Employing various biochemical techniques, including Surface Plasmon Resonance, low molecular weight HS microarray screening, and serial HS mutant cell surface binding assays, we demonstrate TREM2's robust affinity for HS, and the effective binding requires a minimum HS size of approximately 10 saccharide units. Notably, TREM2 selectively binds specific HS structures, with 6-O-sulfation and, to a lesser extent, the iduronic acid residue playing crucial roles. N-sulfation and 2-O-sulfation are dispensable for this interaction. Furthermore, we reveal that 6-O-sulfation is essential for HS-TREM2 ternary complex formation on the microglial cell surface, and HS and its 6-O-sulfation are necessary for TREM2-mediated ApoE3 uptake in microglia. By delineating the interaction between HS and TREM2 on the microglial cell surface and demonstrating its role in facilitating TREM2-mediated ApoE uptake by microglia, our findings provide valuable insights that can inform targeted interventions for modulating microglial functions in Alzheimer's disease.

Keywords:  TREM2; binary complex; heparan sulfate; microglia; structure-function

DOI:  https://doi.org/10.1016/j.jbc.2024.107691
ACS Appl Bio Mater. 2024 Aug 19.

Delivery of Nitric Oxide by Chondroitin Sulfate C Increases the Rate of Wound Healing through Immune Modulation.

Samantha L Picciotti, Heba El-Ahmad, Madelyn P Bucci, Quincy E Grayton, Shannon M Wallet, Mark H Schoenfisch.

  Chronic wounds impact 2.5% of the United States population and will continue to be a major clinical challenge due to increases in population age, chronic disease diagnoses, and antibiotic-resistant infection. Nitric oxide (NO) is an endogenous signaling molecule that represents an attractive, simple therapeutic for chronic wound treatment due to its innate antibacterial and immunomodulatory function. Unfortunately, modulating inflammation for extended periods by low levels of NO is not possible with NO gas. Herein, we report the utility of a NO-releasing glycosaminoglycan biopolymer (GAG) for promoting wound healing. GAGs are naturally occurring biopolymers that are immunomodulatory and known to be involved in the native wound healing process. Thus, the combination of NO and GAG biopolymers represents an attractive wound therapeutic due to these known independent roles. The influence and contribution of chondroitin sulfate C (CSC) modified to facilitate controlled and targeted delivery of NO (CSC-HEDA/NO) was evaluated using in vitro cell proliferation and migration assays and an in vivo wound model.

Keywords:  chondroitin sulfate; immune modulation; molecular mechanisms; nitric oxide; wound healing

DOI:  https://doi.org/10.1021/acsabm.4c00731
Carbohydr Polym. 2024 Nov 01. pii: S0144-8617(24)00681-7. [Epub ahead of print]343 122455

In situ formed aldehyde-modified hyaluronic acid hydrogel with polyelectrolyte complexes of aldehyde-modified chondroitin sulfate and gelatin: An approach for minocycline delivery.

Tutut Habibah, Jana Matonohová, Jaromír Kulhánek, Una Fitzgerald, Marek Ingr, Martin Pravda, Abhay Pandit, Vladimír Velebný.

  Polysaccharides like hyaluronan (HA) and chondroitin sulfate (CS) are native of the brain's extracellular matrix crucial for myelination and brain maturation. Despite extensive research on HA and CS as drug delivery systems (DDS), their high water solubility limits their application as drug carriers. This study introduces an injectable DDS using aldehyde-modified hyaluronic acid (HAOX) hydrogel containing polyelectrolyte complexes (PEC) formed with calcium, gelatin, and either CS or aldehyde-modified CS (CSOX) to deliver minocycline for Multiple Sclerosis therapy. PECs with CSOX enable covalent crosslinking to HAOX, creating immobilized PECs (HAOX_PECOX), while those with CS remain unbound (HAOX_PECS). The in situ forming DDS can be administered via a 20 G needle, with rapid gelation preventing premature leakage. The system integrates into an implanted device for minocycline release through either Fickian or anomalous diffusion, depending on PEC immobilization. HAOX_PECOX reduced burst release by 88 %, with a duration of 127 h for 50 % release. The DDS exhibited an elastic modulus of 3800 Pa and a low swelling ratio (0-1 %), enabling precise control of minocycline release kinetics. Released minocycline reduced IL-6 secretion in the Whole Blood Monocytes Activation Test, suggesting that DDS formation may not alter the biological activity of the loaded drug.

Keywords:  Chondroitin sulphate; Hyaluronic acid; Hydrogel; Minocycline; Polyelectrolyte complexes

DOI:  https://doi.org/10.1016/j.carbpol.2024.122455
Heliyon. 2024 Aug 15. 10(15): e35032

Analysis of indoxyl sulfate in biological fluids with emphasis on sample preparation techniques: A comprehensive analytical review.

Samira Shafiee, Siavoush Dastmalchi, Afshin Gharekhani, Ali Shayanfar.

  The uremic toxin indoxyl sulfate (IS) has been related to the development of various medical conditions notably chronic kidney disease (CKD). Hence, quantification of this biomarker in biological fluids may be a diagnostic tool to evaluate renal system functionality. Numerous analytical methods including liquid chromatography, gas chromatography, spectroscopy, and electrochemical techniques have since been used to analyze IS in different biological fluids. The current review highlights the relevant studies that assessed IS with a special focus on sample preparation, which is essential to reduce or eliminate the effect of endogenous components from the matrix in bioanalysis.

Keywords:  Bioanalysis; Chronic kidney disease (CKD); Indoxyl sulfate (IS); Sample preparation; Uremic toxin

DOI:  https://doi.org/10.1016/j.heliyon.2024.e35032
J Biomater Sci Polym Ed. 2024 Aug 18. 1-17

Enhanced osteogenesis and antibacterial activity of dual-functional PEEK implants via biomimetic polydopamine modification with chondroitin sulfate and levofloxacin.

Mengjue Li, Junyan Liu, Yutong Li, Wenyu Chen, Zhou Yang, Yayu Zou, Yi Liu, Yue Lu, Jianfei Cao.

  Polyetheretherketone (PEEK) implants have emerged as a clinically favored alternative to titanium alloy implants for cranial bone substitutes due to their excellent mechanical properties and biocompatibility. However, the biological inertness of PEEK has hindered its clinical application. To address this issue, we developed a dual-functional surface modification method aimed at enhancing both osteogenesis and antibacterial activity, which was achieved through the sustained release of chondroitin sulfate (CS) and levofloxacin (LVFX) from a biomimetic polydopamine (PDA) coating on the PEEK surface. CS was introduced to promote cell adhesion and osteogenic differentiation. Meanwhile, incorporation of antibiotic LVFX was essential to prevent infections, which are a critical concern in bone defect repairing. To our delight, experiment results demonstrated that the SPKD/CS-LVFX specimen exhibited enhanced hydrophilicity and sustained drug release profiles. Furthermore, in vitro experiments showed that cell growth and adhesion, cell viability, and osteogenic differentiation of mouse calvaria-derived osteoblast precursor (MC3T3-E1) cells were significantly improved on the SPKD/CS-LVFX coating. Antibacterial assays also confirmed that the SPKD/CS-LVFX specimen effectively inhibited the growth of Escherichia coli and Staphylococcus aureus, attributable to the antibiotic LVFX released from the PDA coating. To sum up, this dual-functional PEEK implant showed a promising potential for clinical application in bone defects repairing, providing excellent osteogenic and antibacterial properties through a synergistic approach.

Keywords:  Biomimetic; PEEK; antibacterial property; chondroitin sulfate; levofloxacin; osteogenesis

DOI:  https://doi.org/10.1080/09205063.2024.2390745
Genes Genomics. 2024 Aug 17.

Prognostic significance of HS2ST1 expression in patients with hepatocellular carcinoma.

Ting Ting Chung, Sang Kyum Kim, Seung Jin Lee.

   BACKGROUND: Heparan sulfate 2-O-sulfotransferase 1 (HS2ST1) catalyzes the sulfation of glucuronic acid residues in heparan sulfate proteoglycans, enabling these proteoglycans to interact with numerous ligands within tumor microenvironments. However, the prognostic role of HS2ST1 expression in cancer remains unclear.
OBJECTIVE: This investigated HS2ST1 expression levels and their prognostic significance in various cancer types, demonstrated the prognostic value of HS2ST1 expression in hepatocellular carcinoma (HCC) patients, and identified molecular signatures associated with HS2ST1 expression.
METHODS: HS2ST1 expression and patient survival data from The Cancer Genome Atlas (TCGA) datasets were analyzed using the Gene Expression Profiling Interactive Analysis (GEPIA) portal. We obtained gene expression and clinicopathological information on HCC patients from the TCGA and the Japan and France International Cancer Genome Consortium (ICGC) databases and performed survival analyses. We also examined relevant protein networks, differentially expressed genes, gene set enrichments, and tumor immune microenvironment features associated with HS2ST1 expression.
RESULTS: HS2ST1 exhibited higher expression in eight tumor types compared with normal tissues and was associated with poor prognoses in five tumors, including HCC. HS2ST1 status correlated with poor prognosis in two ICGC HCC cohorts. Elevated HS2ST1 expression in HCC tumors was associated with signaling pathways involved in cell cycle progression, protein secretion, and mTORC1 signaling. Moreover, HS2ST1 expression levels were inversely correlated with immune cell infiltration in the tumor microenvironment.
CONCLUSION: Our study elucidates the prognostic significance of HS2ST1 expression in HCC patients and provides insights into the potential roles of HS2ST1 in signaling pathways and the tumor microenvironment.

Keywords:  GSEA; HS2ST1; Heparan sulfate sulfotransferase; Hepatocellular carcinoma; ICGC; Prognosis; TCGA

DOI:  https://doi.org/10.1007/s13258-024-01556-0
Glycobiology. 2024 Aug 22. pii: cwae065. [Epub ahead of print]

Sialylated keratan sulfates on MUC5B are Siglec-8 ligands in the human esophagus.

T August Li, Anabel Gonzalez-Gil, Abduselam K Awol, Steven J Ackerman, Benjamin C Orsburn, Ronald L Schnaar.

  Human sialic acid-binding immunoglobulin-like lectins (Siglecs) are expressed on subsets of immune cells. Siglec-8 is an immune inhibitory Siglec on eosinophils and mast cells, which are effectors in allergic disorders including eosinophilic esophagitis. Inhibition occurs when Siglec-8 is crosslinked by multivalent Siglec ligands in target tissues. Previously we discovered a high-affinity Siglec-8 sialoglycan ligand on human airways composed of terminally sialylated keratan sulfate chains carried on a single protein, DMBT1. Here we extend that approach to another allergic inflammatory target tissue, human esophagus. Lectin overlay histochemistry revealed that Siglec-8 ligands are expressed predominantly by esophageal submucosal glands, and are densely packed in submucosal ducts leading to the lumen. Expression is tissue-specific; esophageal glands express Siglec-8 ligand whereas nearby gastric glands do not. Extraction and resolution by gel electrophoresis revealed a single predominant human esophageal Siglec-8 ligand migrating at >2 MDa. Purification by size exclusion and affinity chromatography, followed by proteomic mass spectrometry, revealed the protein carrier to be MUC5B. Whereas all human esophageal submucosal cells express MUC5B, only a portion convert it to Siglec-8 ligand by adding terminally sialylated keratan sulfate chains. We refer to this as MUC5BS8L. Material from the esophageal lumen of live subjects revealed MUC5BS8L species ranging from ~1-4 MDa. We conclude that MUC5B in the human esophagus is a protein canvas on which Siglec-8 binding sialylated keratan sulfate chains are post-translationally added. These data expand understanding of Siglec-8 ligands and may help us understand their roles in allergic immune regulation.

Keywords:  Siglec; keratan sulfate; mucin; sialic acid; submucosal gland

DOI:  https://doi.org/10.1093/glycob/cwae065
J Sep Sci. 2024 Aug;47(16): e2400436

Substantial benefits of an inert biphenyl column for the analysis of steroids and their phase II metabolites in biological samples.

Mathieu Galmiche, Marie-Anaïs Monat, Diego A Lopez, Cyrille Lamboley, Paul Connolly, Sergey Girel, Davy Guillarme, Isabel Meister, Serge Rudaz.

  Steroids can be used as biomarkers in clinical metabolomics and other fields related to human toxicology. This chemical group is known for its complexity, considering its number of isobaric compounds and the wide variety of phases I and II metabolic pathways that parent compounds can undergo. For a successful analysis of steroids in biological samples, liquid chromatography separation must be finely tuned. It is especially challenging for glucuronidated and sulfated steroids derivatives that bear polar heads and can be affected by non-specific adsorption. The benefits of a biphenyl stationary phase chemistry for the selectivity of the separation of steroids and their phase II metabolites and the extent to which nonspecific adsorption phenomena could degrade chromatographic performance were investigated. Replacing a conventional hardware by a passivated hardware allowed to considerably reduce peaks width and asymmetry of sulfated species. The addition of weak ion pairing agents in the mobile phase could also help to reduce non-specific adsorption but are detrimental to mass spectrometry detection. As confirmed by the successful detection of 52 steroids in plasma, the use of a biphenyl stationary phase complemented by a passivated column hardware is of great help for a successful biomedical analysis of steroids and their phase II metabolites.

Keywords:  glucuronides; liquid chromatography; non‐specific adsorption; steroids; sulfates

DOI:  https://doi.org/10.1002/jssc.202400436
J Hazard Mater. 2024 Aug 17. pii: S0304-3894(24)02138-1. [Epub ahead of print]478 135559

Hexavalent chromium uptake in rice (Oryza sativa L.) mediated by sulfate and phosphate transporters OsSultr1;2 and OsPht1;1.

Jingjing Li, Weipeng Xie, Hua Qi, Shengsheng Sun, Tenghaobo Deng, Yetao Tang, Rongliang Qiu.

  Chromium (Cr) soil contamination is a critical global environmental concern, with hexavalent chromium (Cr[VI]) being especially perilous due to its high mobility, bioavailability, and phytotoxicity. This poses a significant threat to the cultivation of crops, particularly rice, where the mechanisms of Cr(VI) absorption remain largely unexplored. This study uncovered a competitive interaction between Cr(VI) and essential nutrients-sulfate and phosphate during the uptake process. Notably, deficiencies in sulfate and phosphate were associated with a marked increase in Cr(VI) accumulation in rice, reaching up to 76.5 % and 77.7 %, respectively. Employing q-PCR, this study identified significant up-regulation of the sulfate transporter gene, OsSultr1;2, and the phosphate transporter gene, OsPht1;1, in response to Cr(VI) stress. Genetic knockout studies have confirmed the crucial role of OsSultr1;2 in Cr(VI) uptake, with its deletion leading to a 36.1 % to 69.6 % decrease in Cr uptake by rice roots. Similarly, the knockout of OsPht1;1 resulted in an 18.1 % to 25.7 % decrease in root Cr accumulation. These findings highlight the key role of the sulfate transporter OsSultr1;2 in Cr(VI) uptake, with phosphate transporters also contributing significantly to the process. These insights are valuable for developing rice varieties with reduced Cr(VI) accumulation, ensuring the safety of rice grain production.

Keywords:  Cr(VI); Phosphate; Rice; Sulfate; Uptake

DOI:  https://doi.org/10.1016/j.jhazmat.2024.135559
RSC Adv. 2024 Aug 16. 14(36): 26596-26603

Electrospun EVOH/AST-120 hybrid nanofiber membranes for removal of indoxyl sulfate from blood.

Makoto Sasaki, Rieko Hirata, Ayano Konagai, Mitsuhiro Ebara.

Nanofibers containing activated carbon using poly(ethylene-co-vinyl alcohol) (EVOH) were prepared to remove indoxyl sulfate (IS) from the blood. IS is a urinary toxin that is highly toxic and triggers the progression of chronic kidney disease (CKD). Here, nanofibers containing activated carbon (AST-120), which has been used practically as an adsorbent for indole (a precursor of IS), were fabricated via electrospinning for the adsorption and removal of IS from the blood. EVOH containing different ethylene ratios was used as the nanofiber material; moreover, the effect of the ethylene ratio on various properties of the nanofibers, such as surface wettability and the IS adsorption rate, was investigated. As a result, EVOH/AST-120 nanofibers comprising EVOH with a low ethylene ratio exhibited faster IS adsorption behavior. This adsorption behavior agreed well with the pseudo-second-order model, suggesting that the diffusion of IS into the nanofibers is the rate-limiting step of the process of adsorption. Furthermore, the nanofibers successfully reduced the IS concentration in the blood under circulating conditions. Therefore, these EVOH/AST-120 nanofibers are expected to greatly improve the prognosis of patients with CKD when used in combination with the current hemodialysis therapy as an IS-adsorbing filter.

DOI: https://doi.org/10.1039/d4ra04501g