bims-supasi 2025-01-05 papers

bims-supasi

Biomed News

on Sulfation pathways and signalling

Issue of 2025–01–05
seven papers selected by
Jonathan Wolf Mueller, University of Birmingham

Basic Science and Pathogenesis.
Evolution of SARS-CoV-2 spike trimers towards optimized heparan sulfate cross-linking and inter-chain mobility.
Basic Science and Pathogenesis.
Garlic (Allium sativum L.) Organosulfur Compounds Inhibit Breast Cancer Cell Proliferation by Decreasing Steroid Sulfatase Levels.
Association between serum total indoxyl sulfate, intraperitoneal inflammation, and peritoneal dialysis technique failure: a 3-year prospective cohort study.
Liquid chromatography coupled with high resolution mass spectrometry reveals the inhibitory effects of Huangkuisiwu formula on biosynthesis of protein-binding uremic toxins in rats with chronic kidney disease.
Conductive polyacrylamide/pullulan/ammonium sulfate hydrogels with high toughness, low-hysteresis and tissue-like modulus as flexible strain sensors.

Alzheimers Dement. 2024 Dec;20 Suppl 1 e088949

Basic Science and Pathogenesis.

Chunyu Wang, Jian Liu, Lianchun Wang, Dylan Mah, Fuming Zhang.

BACKGROUND: Heparan sulfate (HS) interacts with many important proteins. These interactions are primarily driven by electrostatics, with specificity determined by sulfation patterns. Although 3-O-sulfation is a rare modification in HS, several genome-wide association studies (GWAS) revealed that the Hs3st1 gene, encoding HS-3-O-sulfotransferase-1, is significantly linked to late onset AD risk.
METHOD: NMR, SPR, glycan array were used to study HS-protein interactions. A novel LC-MS/MS method was developed to quantify the low abundance 3-O-sulfated HS from human brain, with 13C labeled calibrants. Alexa-488 labeled Tau and ApoE was used to assess tau cell surface binding and uptake in HS3ST-1 knockout cellular lines.
RESULT: We have demonstrated that tau, the major component of neurofibrillary tangles, specifically recognizes the 3-O-sulfation (3-OS). In addition, ApoE, whose isoform ApoE4 is the most significant genetic risk factor for late onset AD, also interacts with 3-OS. In addition, the affinity between ApoE isoform and heparin correlates with AD risk, with ApoE4 having the highest affinity to heparin, while the protective ApoE2 and Christchurch having significantly lower affinity. With a novel LC-MS/MS method for quantifying 3-O-sulfation, our recent studies have further demonstrated a marked increase in 3-O-sulfated HS in AD brains.
CONCLUSION: Taken together, our findings underscore the importance of 3-OS in AD pathogenesis, suggesting its potential as a novel drug target and biomarker in Alzheimer's disease.

DOI: https://doi.org/10.1002/alz.088959
Sci Rep. 2024 Dec 31. 14(1): 32174

Evolution of SARS-CoV-2 spike trimers towards optimized heparan sulfate cross-linking and inter-chain mobility.

Jurij Froese, Marco Mandalari, Monica Civera, Stefano Elli, Isabel Pagani, Elisa Vicenzi, Itzel Garcia-Monge, Daniele Di Iorio, Saskia Frank, Antonella Bisio, Dominik Lenhart, Rudolf Gruber, Edwin A Yates, Ralf P Richter, Marco Guerrini, Seraphine V Wegner, Kay Grobe.

  The heparan sulfate (HS)-rich extracellular matrix (ECM) serves as an initial interaction site for the homotrimeric spike (S) protein of SARS-CoV-2 to facilitate subsequent docking to angiotensin-converting enzyme 2 (ACE2) receptors and cellular infection. More recent variants, notably Omicron, have evolved by swapping several amino acids to positively charged residues to enhance the interaction of the S-protein trimer with the negatively charged HS. However, these enhanced interactions may reduce Omicron's ability to move through the HS-rich ECM to effectively find ACE2 receptors and infect cells, raising the question of how to mechanistically explain HS-associated viral movement. In this work, we show that Omicron S proteins have evolved to balance HS interaction stability and dynamics, resulting in enhanced mobility on an HS-functionalized artificial matrix. This property is achieved by the ability of Omicron S-proteins to cross-link at least two HS chains, allowing direct S-protein switching between chains as a prerequisite for cell surface mobility. Optimized HS interactions can be targeted pharmaceutically, as an HS mimetic significantly suppressed surface binding and cellular infection specifically of the Omicron variant. These findings suggest a robust way to interfere with SARS-CoV-2 Omicron infection and potentially future variants.

Keywords:  COVID-19; Heparan sulfate; Heparin; Intersegmental transfer; S-protein; SARS-CoV-2

DOI:  https://doi.org/10.1038/s41598-024-84276-5
Alzheimers Dement. 2024 Dec;20 Suppl 1 e086006

Basic Science and Pathogenesis.

Barbara E Stopschinski, Sandi-Jo Estill-Terpack, Sushobhna Batra, Shubhangi Pandey, Jaime Vaquer-Alicea Iii, Marc I Diamond.

BACKGROUND: The prion model of tau propagation in Alzheimer's Disease predicts that tau seeds are released from cells and taken up by neighboring cells, resulting in spreading of the disease. Our previous work revealed that tau aggregates bind to heparan sulfate proteoglycans (HSPGs) on the cell surface, followed by cellular uptake via macropinocytosis. HSPGs are glycoproteins, consisting of a protein core and decorated with linear glycosaminoglycan (GAG) chains called heparan sulfate (HS) with highly variable sulfation patterns. We have previously used a combination of biochemical, pharmacological and genetic tools to demonstrate that N-sulfation (mediated by the N-sulfotransferase NDST1) of the HS chain is critical for tau uptake in cell models. This project is a stepwise approach to validate HSPGs and specifically NDST1 as a therapeutic target to block tau spread in vivo.
METHOD: We induced NDST1 knockout using CRISPR/Cas9 followed by rescue experiments and tau uptake assays in HEK293T cells. Tau uptake assays with heparin were conducted in murine hippocampal neurons and astrocytes. We employed stereotactic adeno-associated virus 8 (AAV8)-mediated intracerebral delivery of small hairpin RNA (shRNA) to target NDST1 in PS19 mice, a tauopathy mouse model (collaboration with Merck).
RESULT: Our data demonstrates that reduction of the N-deacetylase activity with selected point mutations within NDST1 can decrease tau uptake substantially (Figure 1). Our experiments in primary murine neurons and astrocytes suggest that HSPGs are crucial for tau uptake in these cell types (Figure 2). NDST1 knockdown with shRNA in PS19 mice achieved >75% reduction of messenger RNA (mRNA) levels and was non-toxic (Figure 3).
CONCLUSION: Our findings suggest that NDST1 is a potentially druggable target, and that the HSPG pathway plays a role for tau uptake in multiple cell types. We worked with GemPharmatech for the generation of a floxed NDST1 mouse line. We plan to breed the floxed NDST1 mice to PS19 mice and selected neuronal, astrocytic and microglial Cre driver lines. We anticipate that knockout of NDST1 in vivo will reduce tau spread in mouse brain. If successful, our study will inspire multiple lines of follow up work such as the design of enzymatic inhibitors and genetic silencing tools.

DOI: https://doi.org/10.1002/alz.086006
Anticancer Res. 2025 Jan;45(1): 145-152

Garlic (Allium sativum L.) Organosulfur Compounds Inhibit Breast Cancer Cell Proliferation by Decreasing Steroid Sulfatase Levels.

Akira Sato, Ayano Yabuki, Genta Sato, Hina Nemoto, Yuta Ogawa, Makoto Ohira.

   BACKGROUND/AIM: Breast cancer is mostly affected by estrogen, which promotes proliferation, tumorigenesis, and cancer progression. Estrogen sulfotransferase (SULT1E1) catalyzes sulfation to inactivate estrogens, whereas steroid sulfatase (STS) catalyzes estrogen sulfate hydrolysis to activate estrogens in breast cancer cells. Three major organosulfur compounds in garlic (Allium sativum L.), diallyl sulfide (DAS), diallyl disulfide (DADS), and diallyl trisulfide (DATS), are known to exert anticancer effects against breast cancer. This study aimed to investigate the effects of these compounds on proliferation and SULT1E1 and STS protein levels in breast cancer cells.
MATERIALS AND METHODS: Cell proliferation and SULT1E1 and STS protein levels in MCF-7 breast cancer cells treated with DAS, DADS, and DATS were analyzed via 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyltetrazolium bromide and western blotting assays, respectively.
RESULTS: DADS and DATS concentration-dependently inhibited MCF-7 cell proliferation. Specifically, DATS, followed by DADS and DAS (each 100 μmol/l), demonstrated the most significant inhibition of cell proliferation. DADS and DATS also decreased the STS protein levels. Notably, DAS, DADS, and DATS did not affect the SULT1E1 protein levels. In MCF-7 cells treated with DAS, DADS, and DATS, cell proliferation was positively correlated with STS protein expression.
CONCLUSION: Overall, our findings highlight the potential of DADS and DATS as promising agents for preventing and treating breast cancer by decreasing STS protein expression and suppressing active estrogen levels in breast cancer cells.

Keywords:  Diallyl disulfide; breast cancer; cell proliferation; diallyl trisulfide; steroid sulfatase; sulfotransferase

DOI:  https://doi.org/10.21873/anticanres.17401
BMC Nephrol. 2024 Dec 31. 25(1): 475

Association between serum total indoxyl sulfate, intraperitoneal inflammation, and peritoneal dialysis technique failure: a 3-year prospective cohort study.

Natalia Stepanova, Victoria Driianska, Lesya Korol, Lyudmyla Snisar.

   BACKGROUND: The impact of protein-bound uremic toxins, specifically indoxyl sulfate (IS) on peritoneal dialysis (PD) complications remains controversial. This study aimed to explore the link between serum total IS (tIS) levels, proinflammatory cytokines in serum and peritoneal dialysis effluent (PDE), and PD technique survival.
METHODS: In this prospective cohort study, 84 patients were followed up for three years and analyzed. Stratification into low-tIS (< 22.6 µmol/L) and high-tIS (≥ 22.6 µmol/L) groups was based on the median serum tIS concentration. Logistic regression, Kaplan-Meier, receiving operation characteristic, and Cox regression analyses assessed associations between tIS levels, cytokine concentrations (IL-6, MCP-1, TNF-α), and PD technique failure.
RESULTS: Patients in the high-tIS group were older and had a higher prevalence of diabetes, a greater incidence of PD-related peritonitis, elevated diastolic blood pressure, and lower HDL cholesterol compared to those in the low-tIS group. They also exhibited higher peritoneal transport characteristics, lower dialysis adequacy, and reduced peritoneal creatinine clearance. Elevated tIS levels significantly correlated with higher PDE cytokine levels, without a corresponding rise in serum cytokine levels. Serum tIS levels ≥ 50 µmol/L predicted PD technique failure with 70.4% sensitivity and 87.9% specificity (p < 0.0001). The association between high tIS levels and PD technique failure remained significant after adjusting for confounders identified in logistic regression, including peritoneal weekly creatinine clearance, the D/P creatinine ratio, high peritoneal transport status, and PDE IL-6 and MCP-1 concentrations (HR 2.9, 95% CI 1.13; 8.21).
CONCLUSION: Our findings are the first to demonstrate a link between elevated tIS levels, peritoneal inflammation, and an increased risk of PD technique failure. Monitoring tIS levels in PD patients could be clinically relevant for risk assessment and personalized management, potentially improving long-term PD outcomes. Future research should explore interventions targeting tIS reduction to alleviate peritoneal inflammation and improve PD prognosis.

Keywords:  Cytokines; Dialysis technique failure; Peritoneal dialysis; Total indoxyl sulfate

DOI:  https://doi.org/10.1186/s12882-024-03935-x
J Chromatogr B Analyt Technol Biomed Life Sci. 2024 Dec 28. pii: S1570-0232(24)00454-9. [Epub ahead of print]1252 124445

Liquid chromatography coupled with high resolution mass spectrometry reveals the inhibitory effects of Huangkuisiwu formula on biosynthesis of protein-binding uremic toxins in rats with chronic kidney disease.

Jianping Li, Yumeng Wang, Chengxi Li, Jinao Duan, Jianjing Liu, Jianming Guo.

  Chronic kidney disease (CKD) is recognized as a common disorder worldwide. Protein-binding uremic toxins that cannot be efficiently removed by extracorporeal renal replacement therapies, such as indoxyl sulfate (IS) and p-cresyl sulfate (PCS), are associated with high risks of cardiovascular complications and high mortality in CKD population. This study aimed to explore the therapeutical effects of Huangkuisiwu formula (HKSWF) on CKD rats. Moreover, the underlying mechanisms of HKSWF to inhibit the biosynthesis of IS and PCS were studied. Untargeted metabolomics based on UHPLC-QTOF/MS was conducted to analyze the alterations of endogenous metabolites in plasma. Levels of IS and PCS in plasma and peripheral tissues, as well as levels of amino acids in colonic contents were analyzed by UHPLC-TQ/MS. Levels of indole and p-cresol, the precursors of IS and PCS, in feces and colonic contents were quantified by HPLC-FLD. mRNA and protein expression of sulfotransferase 1 a1 (SULT1A1) were determined by qPCR and Western blotting, respectively. The ability of colonic microbiota to metabolize amino acids into precursors, as well as the activity of sulfotransferase to catalyze precursors into uremic toxins were evaluated by detecting corresponding products from specific substrates. 16S rRNA sequencing were conducted to analyze the profile of gut microbiota. The results showed that HKSWF significantly alleviated the structural and functional impairment of kidney, as well as improved the global metabolic disorders in CKD rats. IS and PCS were identified as the key differential metabolites that contributed to the effects of HKSWF. HKSWF significantly reduced the levels of IS and PCS in plasma, kidney, liver and heart of CKD rats. HKSWF showed no significant effects on the expression of SULT1A1 or the activity of sulfotransferase. HKSWF significantly decreased the levels of indole and p-cresol in the colonic contents and feces of CKD rats, by inhibiting the ability of colonic microbiota to metabolize tryptophan and tyrosine into indole and p-cresol. Alterations in the profile of amino acids and gut microbiota in CKD rats were significantly improved by HKSWF treatment. Conclusively, HKSWF inhibited gut-microbiota mediated biosynthesis of indole and p-cresol, to alleviate the accumulation of IS and PCS in CKD rats.

Keywords:  Chronic kidney disease; Gut microbiota; Huangkuisiwu formula; Indoxyl sulfate; Protein-binding uremic toxin; p-Cresyl sulfate

DOI:  https://doi.org/10.1016/j.jchromb.2024.124445
Int J Biol Macromol. 2024 Dec 26. pii: S0141-8130(24)09994-X. [Epub ahead of print]291 139183

Conductive polyacrylamide/pullulan/ammonium sulfate hydrogels with high toughness, low-hysteresis and tissue-like modulus as flexible strain sensors.

Hongyao Ding, Bingyao Wang, Xu Yang, Jie Liu, Wei Sang, Xuewen Li, Yuefang Wen, Hui Li, Xiaodong Shen.

  Conductive hydrogels have great potential for applications in flexible wearable sensors due to the combination of biocompatibility, mechanical flexibility and electrical conductivity. However, constructing conductive hydrogels with high toughness, low hysteresis and skin-like modulus simultaneously remains challenging. In the present study, we prepared a tough and conductive polyacrylamide/pullulan/ammonium sulfate hydrogel with a semi-interpenetrating network. Ammonium sulfate promoted the formation of low-energy-dissipating motifs between polymer chains, reinforcing the gel matrix and resulting in excellent mechanical properties, including a high stretchability of 2063 %, a high strength of 890 kPa, and a high toughness of 4268 kJ/m3. The hydrogen bonds formed within the network endowed the gels with low-hysteresis under deformation. The unique semi-interpenetrating network structure provided the gels with a tissue-like low modulus. Additionally, the resulting hydrogels exhibited a high conductivity of 2.39 S/m and excellent anti-freezing properties, making them suitable for flexible strain sensors. These sensors demonstrated high sensitivity over a broad strain window of 0.1-1500 %, enabling the detection of various human motions and the recognition of different languages. These findings emphasize the potential of the composite hydrogels as wearable strain sensors for flexible devices.

Keywords:  High toughness; Low-hysteresis; Skin-like modulus

DOI:  https://doi.org/10.1016/j.ijbiomac.2024.139183