bims-supasi 2023-03-26 papers

bims-supasi

Biomed News

on Sulfation pathways and signalling

Issue of 2023–03–26
eight papers selected by
Jonathan Wolf Mueller, University of Birmingham

Antiviral activity of marine sulfated glycans against pathogenic human coronaviruses.
Low sulfated heparan sulfate mimetic differentially affects repair in immune-mediated and toxin-induced experimental models of demyelination.
Structure, anti-SARS-CoV-2, and anticoagulant effects of two sulfated galactans from the red alga Botryocladia occidentalis.
Urinary Glycosaminoglycans Are Associated with Recurrent UTI and Urobiome Ecology in Postmenopausal Women.
Inflammatory stimuli induce shedding of heparan sulfate from arterial but not venous porcine endothelial cells leading to differential proinflammatory and procoagulant responses.
Assessing Dehydroepiandrosterone Sulfate (DHEAS) as a Novel Biomarker for Monitoring Estrus and Successful Reproduction in Polar Bears.
Nucleation of fibronectin fibril assembly requires binding between heparin and the 13thtype III module of fibronectin.
ZnF3, a sulfated polysaccharide from Antrodia cinnamomea, inhibits lung cancer cells via induction of apoptosis and activation of M1-like macrophage-induced cell death.

Sci Rep. 2023 Mar 23. 13(1): 4804

Antiviral activity of marine sulfated glycans against pathogenic human coronaviruses.

Mary Zoepfl, Rohini Dwivedi, Seon Beom Kim, Michael A McVoy, Vitor H Pomin.

Great interest exists towards the discovery and development of broad-spectrum antivirals. This occurs due to the frequent emergence of new viruses which can also eventually lead to pandemics. A reasonable and efficient strategy to develop new broad-spectrum antivirals relies on targeting a common molecular player of various viruses. Heparan sulfate is a sulfated glycosaminoglycan present on the surface of cells which plays a key role as co-receptor in many virus infections. In previous work, marine sulfated glycans (MSGs) were identified as having antiviral activities. Their mechanism of action relies primarily on competitive inhibition of virion binding to heparan sulfate, preventing virus attachment to the cell surface prior to entry. In the current work we used pseudotyped lentivirus particles to investigate in a comparative fashion the inhibitory properties of five structurally defined MSGs against SARS-CoV-1, SARS-CoV-2, MERS-CoV, and influenza A virus (IAV). MSGs include the disaccharide-repeating sulfated galactan from the red alga Botryocladia occidentalis, the tetrasaccharide-repeating sulfated fucans from the sea urchin Lytechinus variegatus and from the sea cucumber Isostichopus badionotus, and the two marine fucosylated chondroitin sulfates from the sea cucumbers I. badionotus and Pentacta pygmaea. Results indicate specificity of action against SARS-CoV-1 and SARS-CoV-2. Curiously, the MSGs showed decreased inhibitory potencies against MERS-CoV and negligible action against IAV. Among the five MSGs, the two sulfated fucans here studied deserve further attention since they have the lowest anticoagulant effects but still present potent and selective antiviral properties.

DOI: https://doi.org/10.1038/s41598-023-31722-5
Glia. 2023 Mar 21.

Low sulfated heparan sulfate mimetic differentially affects repair in immune-mediated and toxin-induced experimental models of demyelination.

Susan L Lindsay, George A McCanney, Jiangshan Zhan, Miriam Scheld, Rebecca Sherrard Smith, Carl S Goodyear, Edwin A Yates, Markus Kipp, Jeremy E Turnbull, Susan C Barnett.

  There is an urgent need for therapies that target the multicellular pathology of central nervous system (CNS) disease. Modified, nonanticoagulant heparins mimic the heparan sulfate glycan family and are known regulators of multiple cellular processes. In vitro studies have demonstrated that low sulfated modified heparin mimetics (LS-mHeps) drive repair after CNS demyelination. Herein, we test LS-mHep7 (an in vitro lead compound) in experimental autoimmune encephalomyelitis (EAE) and cuprizone-induced demyelination. In EAE, LS-mHep7 treatment resulted in faster recovery and rapidly reduced inflammation which was accompanied by restoration of animal weight. LS-mHep7 treatment had no effect on remyelination or on OLIG2 positive oligodendrocyte numbers within the corpus callosum in the cuprizone model. Further in vitro investigation confirmed that LS-mHep7 likely mediates its pro-repair effect in the EAE model by sequestering inflammatory cytokines, such as CCL5 which are upregulated during immune-mediated inflammatory attacks. These data support the future clinical translation of this next generation modified heparin as a treatment for CNS diseases with active immune system involvement.

Keywords:  CNS repair; EAE; cuprizone; heparan sulfate; multiple sclerosis; myelination

DOI:  https://doi.org/10.1002/glia.24363
Int J Biol Macromol. 2023 Mar 22. pii: S0141-8130(23)01062-0. [Epub ahead of print] 124168

Structure, anti-SARS-CoV-2, and anticoagulant effects of two sulfated galactans from the red alga Botryocladia occidentalis.

Antim K Maurya, Poonam Sharma, Priyanka Samanta, Anter A Shami, Sandeep K Misra, Fuming Zhang, Reena Thara, Deepak Kumar, Deling Shi, Robert J Linhardt, Joshua S Sharp, Robert J Doerksen, Ritesh Tandon, Vitor H Pomin.

  The structure of the sulfated galactan from the red alga Botryocladia occidentalis (BoSG) was originally proposed as a simple repeating disaccharide of alternating 4-linked α-galactopyranose (Galp) and 3-linked β-Galp units with variable sulfation pattern. Abundance was estimated only for the α-Galp units: one-third of 2,3-disulfation and one-third of 2-monosulfation. Here, we isolated again the same BoSG fractions from the anion-exchange chromatography, obtaining the same NMR profile of the first report. More careful NMR analysis led us to revise the structure. A more complex sulfation pattern was noted along with the occurrence of 4-linked α-3,6-anhydro-Galp (AnGalp) units. Interestingly, the more sulfated BoSG fraction showed slightly reduced in vitro anti-SARS-CoV-2 activities against both wild-type and delta variants, and significantly reduced anticoagulant activity. The BoSG fractions showed no cytotoxic effects. The reduction in both bioactivities is attributed to the presence of the AnGalp unit. Docking scores from computational simulations using BoSG disaccharide constructs on wild-type and delta S-proteins, and binding analysis through competitive SPR assays using blood (co)-factors (antithrombin, heparin cofactor II and thrombin) and four S-proteins (wild-type, delta, gamma, and omicron) strongly support the conclusion about the deleterious impact of the AnGalp unit.

Keywords:  Anti-SARS-CoV-2; Anticoagulation; Botryocladia occidentalis; Structural determination; Sulfated galactan

DOI:  https://doi.org/10.1016/j.ijbiomac.2023.124168
ACS Infect Dis. 2023 Mar 21.

Urinary Glycosaminoglycans Are Associated with Recurrent UTI and Urobiome Ecology in Postmenopausal Women.

Michael L Neugent, Neha V Hulyalkar, Ashwani Kumar, Chao Xing, Philippe E Zimmern, Vladimir Shulaev, Nicole J De Nisco.

  Glycosaminoglycans (GAGs) are linear, negatively charged polysaccharides composed of repeating disaccharide units of uronic acid and amino sugars. The luminal surface of the bladder epithelium is coated with a GAG layer. These urothelial GAGs are thought to provide a protective barrier and serve as a potential interaction site with the urinary microbiome (urobiome). Previous studies have profiled urinary GAG composition in mixed cohorts, but the urinary GAG composition in postmenopausal women remains undefined. To investigate the relationship between GAGs and recurrent urinary tract infection (rUTI), we profiled urinary GAGs in a controlled cohort of postmenopausal women. We found that chondroitin sulfate (CS) is the major urinary GAG in postmenopausal women and that urinary CS was elevated in women with active rUTI. We also associated urinary GAGs with urobiome composition and identified bacterial species that significantly associated with urinary GAG concentration. Corynebacterium amycolatum, Porphyromonas somerae, and Staphylococcus pasteuri were positively associated with heparin sulfate or hyaluronic acid, and bacterial species associated with vaginal dysbiosis were negatively correlated with urinary CS. Altogether, this work defines changes in urinary GAG composition associated with rUTI and identifies new associations between urinary GAGs and the urobiome that may play a role in rUTI pathobiology.

Keywords:  chondroitin sulfate; glycosaminoglycan; heparin sulfate; hyaluronic acid; recurrent urinary tract infection; urobiome

DOI:  https://doi.org/10.1021/acsinfecdis.3c00027
Sci Rep. 2023 Mar 18. 13(1): 4483

Inflammatory stimuli induce shedding of heparan sulfate from arterial but not venous porcine endothelial cells leading to differential proinflammatory and procoagulant responses.

Anastasia Milusev, Alain Despont, Jane Shaw, Robert Rieben, Nicoletta Sorvillo.

Endothelial dysfunction is an early event of vascular injury defined by a proinflammatory and procoagulant endothelial cell (EC) phenotype. Although endothelial glycocalyx disruption is associated with vascular damage, how various inflammatory stimuli affect the glycocalyx and whether arterial and venous cells respond differently is unknown. Using a 3D round-channel microfluidic system we investigated the endothelial glycocalyx, particularly heparan sulfate (HS), on porcine arterial and venous ECs. Heparan sulfate (HS)/glycocalyx expression was observed already under static conditions on venous ECs while it was flow-dependent on arterial cells. Furthermore, analysis of HS/glycocalyx response after stimulation with inflammatory cues revealed that venous, but not arterial ECs, are resistant to HS shedding. This finding was observed also on isolated porcine vessels. Persistence of HS on venous ECs prevented complement deposition and clot formation after stimulation with tumor necrosis factor α or lipopolysaccharide, whereas after xenogeneic activation no glycocalyx-mediated protection was observed. Contrarily, HS shedding on arterial cells, even without an inflammatory insult, was sufficient to induce a proinflammatory and procoagulant phenotype. Our data indicate that the dimorphic response of arterial and venous ECs is partially due to distinct HS/glycocalyx dynamics suggesting that arterial and venous thrombo-inflammatory disorders require targeted therapies.

DOI: https://doi.org/10.1038/s41598-023-31396-z
Gen Comp Endocrinol. 2023 Mar 18. pii: S0016-6480(23)00081-3. [Epub ahead of print] 114276

Assessing Dehydroepiandrosterone Sulfate (DHEAS) as a Novel Biomarker for Monitoring Estrus and Successful Reproduction in Polar Bears.

Monica Brandhuber, Shannon Atkinson, Curry Cunningham, Terri Roth, Erin Curry.

  Polar bears (Ursus maritimus) in the wild are under threat due to climate change, primarily loss of sea ice, and experience poor reproductive success in zoos. The polar bear is a seasonally polyestrous species that exhibits embryonic diapause and pseudopregnancy, complicating characterization of reproductive function. Fecal excretion of testosterone and progesterone have been studied in polar bears, but accurately predicting reproductive success remains difficult. Dehydroepiandrosterone (DHEA) is a steroid hormone precursor correlated with reproductive success in other species, but has not been well studied in the polar bear. The purpose of the present study was to characterize the longitudinal excretion of DHEAS, the sulfated form of DHEA, from zoo-housed polar bears using a validated enzyme immunoassay. Lyophilized fecal samples from parturient females (n = 10), breeding non-parturient females (n = 11), a non-breeding adult female, a juvenile female, and a breeding adult male were investigated. Five of the breeding non-parturient females had been previously contracepted, while six were never contracepted. DHEAS concentrations were closely associated with testosterone concentrations (p < 0.05, rho > 0.57) for all reproductive statuses. Breeding females exhibited statistically significant (p < 0.05) increases in DHEAS concentration on or near breeding dates, which were not observed outside of the breeding season, or in the non-breeding or juvenile animals. Breeding non-parturient females exhibited higher median and baseline DHEAS concentrations than parturient females over the course of the breeding season. Previously contracepted (PC) breeding non-parturient females also exhibited higher season-long median and baseline DHEAS concentrations than non-previously (NPC) contracepted breeding non-parturient females. These findings suggest that DHEA is related to estrus or ovulation in the polar bear, that there is an optimal DHEA concentration window, and concentrations exceeding that threshold may be associated with reproductive dysfunction.

Keywords:  Dehydroepiandrosterone Sulfate; Estrus; Polar Bear; Reproduction

DOI:  https://doi.org/10.1016/j.ygcen.2023.114276
J Biol Chem. 2023 Mar 16. pii: S0021-9258(23)00264-8. [Epub ahead of print] 104622

Nucleation of fibronectin fibril assembly requires binding between heparin and the 13thtype III module of fibronectin.

Benjamin M Lovett, Katherine E Hill, Ellie M Randolph, Luqiong Wang, Jean E Schwarzbauer.

  Fibronectin (FN), a critical component of the extracellular matrix, is assembled into fibrils through a cell-mediated process. Heparan sulfate (HS) binds to the III13 module of FN and fibroblasts lacking this glycosaminoglycan exhibit reduced FN fibril assembly. To determine if HS depends on III13 to control FN assembly, we deleted both III13 alleles in NIH 3T3 cells using the CRISPR-Cas9 system. ΔIII13 cells assembled fewer FN matrix fibrils and less DOC-insoluble FN matrix than wild type cells. Little if any mutant FN matrix was assembled when purified ΔIII13 FN was provided to Chinese hamster ovary (CHO) cells, showing that lack of III13 caused the deficiency in assembly by ΔIII13 cells. Addition of heparin promoted the assembly of wild type FN by CHO cells, but it had no effect on the assembly of ΔIII13 FN. Furthermore, heparin binding stabilized the folded conformation of III13 and prevented it from self-associating with increasing temperature suggesting that stabilization by HS/heparin binding might regulate interactions between III13 and other FN modules. This effect would be particularly important at matrix assembly sites where our data show that ΔIII13 cells require both exogenous wild type FN and heparin in the culture medium to maximize assembly site formation. Our results show that heparin-promoted growth of fibril nucleation sites is dependent on III13. We conclude that HS/heparin binds to III13 to promote and control the nucleation and development of FN fibrils.

Keywords:  extracellular matrix; fibronectin; heparan sulfate; heparin; heparin binding domain; matrix assembly; type III module

DOI:  https://doi.org/10.1016/j.jbc.2023.104622
Int J Biol Macromol. 2023 Mar 21. pii: S0141-8130(23)01038-3. [Epub ahead of print] 124144

ZnF3, a sulfated polysaccharide from Antrodia cinnamomea, inhibits lung cancer cells via induction of apoptosis and activation of M1-like macrophage-induced cell death.

Zhi-Hu Lin, Mei-Kuang Lu, Hung-Chih Lo, Chia-Chuan Chang, Ai-Jung Tseng, Chi-Hsein Chao, Tung-Yi Lin.

  Sulfated polysaccharides (Ac-SPSs) of Antrodia cinnamomea present anti-cancer activity. However, the anti-cancer mechanism of Ac-SPSs is not fully understood and remains largely unexplored. In this study, we identify an Ac-SPS with 7.9 kDa, noted ZnF3, and aim to examine the dual anti-cancer functions of ZnF3 on inhibiting cancer cells and activating macrophages. A biological study shows that ZnF3 inhibits lung cancer cells by inducing subG1 population and apoptosis. ZnF3 downregulates the expression of TGFβ receptor in lung cancer cells. In parallel, ZnF3 activates macrophages via induction of TNF-α and IL-6 secretion, NO production and phagocytosis. ZnF3 activates AKT/mTOR pathway and induces M1 type macrophage polarization. Cancer cells co-cultured with ZnF3-stimulated macrophages, leading to inhibition of lung cancer cells. This study demonstrates that ZnF3 not only directly inhibits cancer cells but also activates macrophages-mediated cytotoxic effect on cancer cells. Moreover, ZnF3 may be a supplement for suppressing lung cancer cells.

Keywords:  Anti-cancer activity; Antrodia cinnamomea; Macrophage; Sulfated polysaccharides

DOI:  https://doi.org/10.1016/j.ijbiomac.2023.124144