bims-supasi Biomed News
on Sulfation pathways and signalling
Issue of 2023‒01‒15
ten papers selected by
Jonathan Wolf Mueller
University of Birmingham

  1. J Biochem. 2023 Jan 05. pii: mvac110. [Epub ahead of print]
      Chondroitin sulfate (CS) chains, a class of sulfated glycosaminoglycan (GAG) polysaccharides, are ubiquitously distributed in extra/pericellular matrices that establish microenvironmental niches to support a multitude of cellular events. Such wide-ranging functions of CS chains are attributable not only to their sulfation pattern-dependent structural divergence, but also to their multiple modes of action. Although it has long been accepted that CS chains act as passive structural scaffolds that often behave as co-receptors and/or reservoirs for various humoral factors, the discovery of cell surface receptor molecules for distinct CS chains has offered insights into a novel mode of CS function as dynamic extra/pericellular signaling ligands. A recent report by Gong et al. (Identification of PTPR-interacting proteins by proximity-labeling assay. J. Biochem. 2021; 169:187-194) also strongly reinforced the physiological importance of CS receptor-mediated signaling pathways. In this commentary, we briefly introduce the functional aspects of CS chains as extra/pericellular signaling molecules.
    Keywords:  CS receptor; chondroitin sulfate; glycosaminoglycan; proteoglycan; sulfation
  2. Int J Biol Macromol. 2023 Jan 05. pii: S0141-8130(23)00047-8. [Epub ahead of print] 123179
      Glycosaminoglycans (GAGs), such as heparin, heparan sulfate and chondroitin sulfate, are playing important roles in various biological processes. Due to the laborious work of organic or enzymatic total synthesis of GAGs, different approaches, including glycopolymers, dendrimers, etc., have been developed to mimic the structures and bioactivities of GAGs, but the syntheses can still be difficult. In the current study, a new format of GAG mimetic structure, supramolecularly assembled polymers, have been easily prepared by mixing fluorenylmethoxycarbonyl-diphenylalanine (Fmoc-FF) and sulfated glyco-modified fluorenylmethoxy derivatives (FGS and FG3S). The self-assembly behavior of these polymers into different structural formats of nanoparticles, nanofibers and macroscopic hydrogels upon adjusted concentrations and composite ratios have been detailed studied. The nanofibers modified with highly sulfated glycol groups (FG3S/Fmoc-FF) showed strong promotion effect for cell proliferation, which efficiency was even similar to that of natural heparin, higher than nanoparticles or non-/low-sulfated glyco-modified nanofibers. Moreover, the supramolecular polymers were further made into hydrogels that capable of 3D cell culture. This study provided a novel and efficient approach for GAG mimicking, showing great potential for tissue engineering related applications.
    Keywords:  3D cell culture; Glycosaminoglycans; Hydrogel; Nanofibers; Supramolecular assembly
  3. J Chromatogr B Analyt Technol Biomed Life Sci. 2022 Dec 21. pii: S1570-0232(22)00483-4. [Epub ahead of print]1215 123578
      Protein-bound uremic retention solutes, such as indole-3-acetic acid, indoxyl sulfate, p-cresol and p-cresol sulfate, are associated with the development of several pathologies, namely renal, cardiovascular, and bone toxicities, due to their potential accumulation in the human body, thus requiring analytical methods for monitoring and evaluation. The present review addresses conventional and advanced sample treatment procedures for sample handling and the chromatographic analytical methods developed for quantification of these compounds in different biological fluids, with particular focus on plasma, serum, and urine. The sample preparation and chromatographic methods coupled to different detection systems are critically discussed, focusing on the different steps involved for sample treatment, namely elimination of interfering compounds present in the sample matrix, and the evaluation of their environmental impact through the AGREEprep tool. There is a clear trend for the application of liquid-chromatography coupled to tandem mass spectrometry, which requires protein precipitation, solid-phase extraction and/or dilution prior to analysis of biological samples. Furthermore, from a sustainability point of view, miniaturized methods resorting to microplate devices are highly recommended.
    Keywords:  Biological samples; Indole-3-acetic acid; Indoxyl sulfate; Protein-bound compounds; p-Cresol; p-Cresol sulfate
  4. J Med Chem. 2023 Jan 12.
      Sulfated glycosaminoglycans (GAGs), or synthetic mimetics thereof, are not favorably viewed as orally bioavailable drugs owing to their high number of anionic sulfate groups. Devising an approach for oral delivery of such highly sulfated molecules would be very useful. This work presents the concept that conjugating cholesterol to synthetic sulfated GAG mimetics enables oral delivery. A focused library of sulfated GAG mimetics was synthesized and found to inhibit the growth of a colorectal cancer cell line under spheroid conditions with a wide range of potencies ( 0.8 to 46 μM). Specific analogues containing cholesterol, either alone or in combination with clinical utilized drugs, exhibited pronounced in vivo anticancer potential with intraperitoneal as well as oral administration, as assessed by ex vivo tertiary and quaternary spheroid growth, cancer stem cell (CSC) markers, and/or self-renewal factors. Overall, cholesterol derivatization of highly sulfated GAG mimetics affords an excellent approach for engineering oral activity.
  5. Int J Mol Sci. 2022 Dec 24. pii: 310. [Epub ahead of print]24(1):
      Endometriosis is a chronic inflammatory disease associated with pelvic pain, infertility, and increased cardiovascular risk. Recent studies suggest a possible role of aldosterone as a pro-inflammatory hormone in the pathogenesis of the disease. Cortisol is also an important mediator of stress reaction, but its role is controversial in endometriosis. The aim of this study was to evaluate aldosterone and cortisol levels and blood pressure values in women with endometriosis. We measured blood pressure, plasma aldosterone, renin, cortisol, and dehydroepiandrosterone sulfate (DHEAS) in 20 women with untreated minimal or mild pelvic endometriosis compared with 20 healthy controls matched for age and body mass index. Aldosterone values were similar in the two groups, while renin was significantly lower and the aldosterone to renin ratio was significantly higher in patients with endometriosis than in controls. Systolic blood pressure was in the normal range, but significantly higher in patients with endometriosis. Morning plasma cortisol was normal, but significantly lower in patients with endometriosis compared with controls, while DHEAS to cortisol ratio was similar in the two groups. These preliminary results are evidence of increased biological aldosterone activity and dysregulation of the hypothalamic-pituitary-adrenal axis in early stages of endometriosis. These alterations could play a role in disease development, suggesting new therapeutic targets for aldosterone receptor blockers.
    Keywords:  aldosterone; cortisol; endometriosis; hypertension; inflammation; mineralocorticoid receptor; spironolactone
  6. Biomacromolecules. 2023 Jan 10.
      Heparin, an anionic biomacromolecule, is routinely used as an anticoagulant during medical surgery to prevent blood clot formation and in the treatment of several heart, lung, and circulatory disorders having a higher risk of blood clotting. We herein report supramolecular polymeric nanoassemblies of cationic pyrene-tagged bis-imidazolium amphiphiles for heparin detection with high sensitivity and selectivity in aqueous buffer, plasma, and serum media. The nano-assemblies exhibited cyan-green excimeric emission in aqueous media, and their multivalent array of positive surface charges allowed them to form co-assemblies with heparin, resulting in significantly enhanced emission. This provided a convenient method for heparin detection in buffer at nanomolar concentrations, and most notably, a ratiometric fluorescence response was obtained even in highly competitive 100% human serum and 100% human plasma in a clinically relevant concentration range. Moreover, using the heparin-based luminescent co-assemblies, protamine sulfate, a clinically administered antidote to heparin, was also detected in 100% human serum and 100% human plasma at sub-micromolar concentrations.
  7. Analyst. 2023 Jan 10.
      Experts typically define vitamin D deficiency levels by the determination of a circulating 25-hydroxyvitamin D3-calcifediol prohormone. A large part of the population is characterized by deficient vitamin D levels (calcifediol < 20 ng mL-1) despite individuals not being affected by any disorder. Cholecalciferol (vitamin D3) and/or calcifediol supplementation is a common practice for vitamin D-deficient individuals as recommended by international scientific societies and official agencies. In the last few years, several studies have reported the presence of conjugated vitamin D3 metabolites, mainly glucuronidation and sulfation derivatives, although simultaneous quantitative measurements involving phase I and II vitamin D metabolites have not been carried out. A quantitative method based on tandem mass spectrometry detection is proposed here for the combined determination of phase I and phase II vitamin D3 metabolites in human serum. As phase I and phase II metabolites are preferentially ionized in different modes, a switching polarity mode was adopted to determine both groups of compounds in serum at high sensitivity levels (pg mL-1). The validation of this proposal was successfully accomplished by following the Center for Drug Evaluation and Research (CDER) guidelines. Its applicability was tested in a cohort of volunteers with mostly deficient baseline levels. Considering the sulfated form of calcifediol, the sum of its concentrations showed sufficient baseline vitamin D levels in all individuals, suggesting that this could be a novel strategy for vitamin D deficiency definition. Therefore, phase II metabolites are proposed to be included when evaluating the vitamin D status since they provide more information about the overall status of the vitamin D endocrine system. Nevertheless, further studies are required to confirm the biological activity of these conjugated metabolites and the suitability of this strategy for the description of vitamin D deficiency.
  8. Int J Biol Macromol. 2023 Jan 09. pii: S0141-8130(23)00088-0. [Epub ahead of print] 123214
      It remains uncertain how brain glycosaminoglycans (GAGs) contribute to the progression of inflammatory disorders like multiple sclerosis (MS). We investigated here neuroinflammation-mediated changes in GAG composition and metabolism using the mouse model of experimental autoimmune encephalomyelitis (EAE) and sham-immunized mice as controls. Cerebellum, mid- and forebrain at different EAE phases were investigated using gene expression analysis (microarray and RT-qPCR) as well as HPLC quantification of CS and hyaluronic acid (HA). The cerebellum was the most affected brain region showing a downregulation of Bcan, Cspg5, and an upregulation of Dse, Gusb, Hexb, Dcn and Has2 at peak EAE. Upregulation of genes involved in GAG degradation as well as synthesis of HA and decorin persisted from onset to peak, and diminished at remission, suggesting a severity-related decrease in CS and increments in HA. Relative disaccharide quantification confirmed a 3.6 % reduction of CS-4S at peak and a normalization during remission, while HA increased in both phases by 26.1 % and 17.6 %, respectively. Early inflammatory processes led to altered GAG metabolism in early EAE stages and subsequent partially reversible changes in CS-4S and in HA. Targeting early modifications in CS could potentially mitigate progression of EAE/MS.
    Keywords:  Chondroitin sulfate; Experimental autoimmune encephalomyelitis; Extracellular matrix; Glycosaminoglycans; Hyaluronic acid; Multiple sclerosis
  9. J Mass Spectrom Adv Clin Lab. 2023 Jan;27 40-48
      Objectives: Highly selective and sensitive multi-analyte methods for the analysis of steroids are attractive for the diagnosis of endocrine diseases. Commercially available kits are increasingly used for this purpose. These methods involve laborious solid phase extraction, and the respective panels of target analytes are incomplete. We wanted to investigate whether an improvement of kit solutions is possible by introducing automated on-line solid phase extraction (SPE) and combining originally separate analyte panels.Methods: Sample preparation was performed using automated on-line SPE on a high-pressure stable extraction column. Chromatographic separation, including isobaric compounds, was achieved using a 0.25 mM ammonium fluoride-methanol gradient on a small particle size biphenyl column. Standard compounds and internal standard mixtures of two panels of a commercially available kit were combined to achieve an optimized and straightforward detection of 15 endogenous steroids. Validation was performed according to the European Medicines Agency (EMA) guidelines with slight modifications.
    Results: Validation was successfully performed for all steroids over a clinically relevant calibration range. Deviations of intra- and inter-assay accuracy and precision results passed the criteria and no relevant matrix effects were detected due to highly effective sample preparation. External quality assessment samples showed the applicability as a routine diagnostic method, which was affirmed by the analyses of anonymized clinical samples.
    Conclusions: It was found possible to complement a commercially available kit for quantitative serum steroid profiling based on isotope dilution LC-MS/MS by implementing automated on-line SPE, thereby improving the practicality and robustness of the measurement procedure.
    Keywords:  A4, Androstendione; ALDO, Aldosterone; APCI, Atmospheric pressure chemical ionization; CAH, Congenital adrenal hyperplasia; CE, Collision energy; CE-IVD, Certified in-vitro-diagnostic device; CV, Coefficient of variation; DHEA, Dehydro-epiandrosterone; DHEA-S, Dehydro-epiandrosterone sulfate; DHT, Dihydrotestosterone; DOC, 11-deoxycorticosterone; E, Cortisone; E2, Estradiol; EMA, European Medicines Agency; EQA, External quality assessment; ESI, Electrospray ionisation; F, Cortisol; IVD, In-vitro-diagnostic; IVDR, EU In vitro Diagnostic Regulation; LC, Liquid chromatography; LC–MS/MS, Liquid chromatography tandem mass spectrometry; LDT, Laboratory developed test; LLOQ, Lower limit of quantification; MRM, Multiple reaction monitoring; On-line solid phase extraction (SPE); P4, Progesterone; QC, Quality control; Robustness; S/N, Signal-to-noise ratio; SID, Stable-isotope dilution; SPE, Solid phase extraction; SST, System suitability test; Serum steroid profiling; Stable-isotope dilution liquid chromatography-tandem mass spectrometry (SID LC-MS/MS); UHPLC, Ultra high performance liquid chromatography; ULOQ, Upper limit of quantification
  10. Hepatology. 2023 Jan 03.
      BACKGROUND AND AIMS: Hepatocellular carcinoma (HCC) is a malignant disease. Compared with tyrosine kinase inhibitors (the classical therapy), immune checkpoint inhibitors are more effective in the treatment of HCC, despite their limited efficacy. Among these restricted factors, exhaustion of tumor-infiltrated lymphocytes (TILs), especially CD8+ T cells, is a core event. We aimed to determine the key factors contributing to CD8+ T cell infiltration in HCC and investigate the underlying mechanisms.APPROACH AND RESULTS: Using machine learning and multiplex immunohistochemistry analysis, we showed that dedicator of cytokinesis protein 2 (DOCK2) was a potential indicator of infiltrated CD8+ T cells in HCC. Using RNA sequencing, flow cytometry analysis, and mouse HCC models, we demonstrated that DOCK2 inactivation accounted for infiltrated CD8+ T cell exhaustion in tumors. Using quasi-targeted metabolomics, mass spectrum, and mass cytometry by time of flight analysis, we found that cholesterol sulfate (CS) synthesized by sulfotransferase 2B1 (SULT2B1) in tumor cells suppressed DOCK2 enzymatic activity of T cells. Through virtual screening, molecular docking simulation, and experiments validation, we demonstrated that tolazamide reversed DOCK2 inactivation-mediated CD8+ T cell exhaustion and enhanced anti-PDL1 antibody + Apatinib immunotherapeutic effects on HCC.
    CONCLUSIONS: This study indicates that DOCK2 controls CD8+ T cell infiltration in HCC, and CS synthesized by SULT2B1 in tumor cells promotes effector T cell exhaustion. The findings suggest that the usage of conventional drugs affects immunotherapy efficacy in HCC patients.