bims-supasi 2023-04-30 papers

bims-supasi

Biomed News

on Sulfation pathways and signalling

Issue of 2023–04–30
twenty papers selected by
Jonathan Wolf Mueller, University of Birmingham

Quantitative 2D 1H, 13C HSQC NMR Spectroscopy for the Determination of Chondroitin Sulfate and Dermatan Sulfate Content in Danaparoid Sodium.
Inhibition of Cytomegalovirus by Pentacta pygmaea Fucosylated Chondroitin Sulfate Depends on Its Molecular Weight.
Compositional Analysis of Glycosaminoglycans in Different Lung Cancer Types-A Pilot Study.
Well-Defined Heparin Mimetics Can Inhibit Binding of the Trimeric Spike of SARS-CoV-2 in a Length-Dependent Manner.
Multi-step Strategies Toward Regioselectively Sulfated M-Rich Alginates.
Efficient endocytosis of the human lactoferrin N-lobe enhances its antiproliferative activity against human cancer cells.
Structural Characterization and Effects on Insulin Resistance of a Novel Chondroitin Sulfate from Halaelurus burgeri Skin.
Bispecific T cell-engager targeting oncofetal chondroitin sulfate induces complete tumor regression and protective immune memory in mice.
In Vitro Evaluation of Aerosol Therapy with Pentamidine-Loaded Liposomes Coated with Chondroitin Sulfate or Heparin for the Treatment of Leishmaniasis.
A branched 2-O sulfated 1,3-/1,4-galactoglucan from Antrodia cinnamomea exhibits moderate antiproliferative and anti-inflammatory activities.
Glycosaminoglycans from the Starfish Lethasterias fusca: Structures and Influence on Hematopoiesis.
The Uremic Toxin Indoxyl Sulfate Accelerates Senescence in Kidney Proximal Tubule Cells.
Detailing Protein-Bound Uremic Toxin Interaction Mechanisms with Human Serum Albumin in the Pursuit of Designing Competitive Binders.
Predictable genetic recruitment for luciferin sulfation in the convergent evolution of bioluminescence.
Between the Devil and the Deep Blue Sea-Resveratrol, Sulfotransferases and Sulfatases-A Long and Turbulent Journey from Intestinal Absorption to Target Cells.
Approaches to Assure Similarity between Pharmaceutical Heparins from Two Different Manufacturers.
Sulfated endospermic nanocellulose crystals prevent the transmission of SARS-CoV-2 and HIV-1.
Correlation between biochemical and clinical hyperandrogenism parameter in polycystic ovary syndrome in relation to age.
Metabolites of sea cucumber sulfated polysaccharides fermented by Parabacteroides distasonis and their effects on cross-feeding.
Proton-Pump Inhibitors and Serum Concentrations of Uremic Toxins in Patients with Chronic Kidney Disease.

Thromb Haemost. 2023 Apr 24.

Quantitative 2D 1H, 13C HSQC NMR Spectroscopy for the Determination of Chondroitin Sulfate and Dermatan Sulfate Content in Danaparoid Sodium.

Cristina Gardini, Giovanni Boccardi, Marco Guerrini, Edwin Kellenbach, Maarten Lunenburg, Jan-Ytzen van der Meer, Annamaria Naggi, Elena Urso.

OBJECTIVE: Danaparoid sodium is a biopolymeric complex drug composed of the most abundant heparan sulfate (HS) followed in descending order by dermatan sulfate (DS) and chondroitin sulfate (CS). This composite nature explains its peculiar antithrombotic and anticoagulant properties and make it particularly advantageous when the risk of heparin-induced thrombocytopenia occurs. A specific control of the danaparoid composition is required by the Ph. Eur. The monograph includes the CS and DS limit contents and describes the method for their quantification through selective enzymatic degradations.
MATERIALS AND METHODS: In this study, a quantitative two-dimensional nuclear magnetic resonance (NMR) method is proposed as a new method suitable for CS and DS quantification. Statistical comparison of the results provided by the analysis of a series of danaparoid samples with both NMR and enzymatic methods highlights a small systematic difference, likely derived from lyase-resistant sequences bearing oxidized terminals. Some modified structures, whose survival to the enzymatic action was confirmed by mass spectrometry, can be detected and quantified by NMR.
CONCLUSION AND RESULTS: The proposed NMR method can serve for the determination of DS and CS contents, is an easy-to-apply method with no dependence from enzymes and standards, and provides extensive structural information on the overall glycosaminoglycans mixture.

DOI: https://doi.org/10.1055/s-0043-1768225
Viruses. 2023 Mar 28. pii: 859. [Epub ahead of print]15(4):

Inhibition of Cytomegalovirus by Pentacta pygmaea Fucosylated Chondroitin Sulfate Depends on Its Molecular Weight.

Poonam Sharma, Rohini Dwivedi, Priya Ray, Jayanti Shukla, Vitor H Pomin, Ritesh Tandon.

  Many viruses attach to host cells by first interacting with cell surface proteoglycans containing heparan sulfate (HS) glycosaminoglycan chains and then by engaging with specific receptor, resulting in virus entry. In this project, HS-virus interactions were targeted by a new fucosylated chondroitin sulfate from the sea cucumber Pentacta pygmaea (PpFucCS) in order to block human cytomegalovirus (HCMV) entry into cells. Human foreskin fibroblasts were infected with HCMV in the presence of PpFucCS and its low molecular weight (LMW) fractions and the virus yield at five days post-infection was assessed. The virus attachment and entry into the cells were visualized by labeling the purified virus particles with a self-quenching fluorophore octadecyl rhodamine B (R18). The native PpFucCS exhibited potent inhibitory activity against HCMV specifically blocking virus entry into the cell and the inhibitory activities of the LMW PpFucCS derivatives were proportional to their chain lengths. PpFucCS and the derived oligosaccharides did not exhibit any significant cytotoxicity; moreover, they protected the infected cells from virus-induced lytic cell death. In conclusion, PpFucCS inhibits the entry of HCMV into cells and the high MW of this carbohydrate is a key structural element to achieve the maximal anti-viral effect. This new marine sulfated glycan can be developed into a potential prophylactic and therapeutic antiviral agent against HCMV infection.

Keywords:  herpesviruses; marine sulfated glycans; virus entry

DOI:  https://doi.org/10.3390/v15040859
Int J Mol Sci. 2023 Apr 11. pii: 7050. [Epub ahead of print]24(8):

Compositional Analysis of Glycosaminoglycans in Different Lung Cancer Types-A Pilot Study.

Domonkos Pál, Gábor Tóth, Simon Sugár, Kata Dorina Fügedi, Dániel Szabó, Ilona Kovalszky, Dávid Papp, Gitta Schlosser, Csaba Tóth, Tamás Tornóczky, László Drahos, Lilla Turiák.

  Lung cancer is one of the most commonly diagnosed cancer types. Studying the molecular changes that occur in lung cancer is important to understand tumor formation and identify new therapeutic targets and early markers of the disease to decrease mortality. Glycosaminoglycan chains play important roles in various signaling events in the tumor microenvironment. Therefore, we have determined the quantity and sulfation characteristics of chondroitin sulfate and heparan sulfate in formalin-fixed paraffin-embedded human lung tissue samples belonging to different lung cancer types as well as tumor adjacent normal areas. Glycosaminoglycan disaccharide analysis was performed using HPLC-MS following on-surface lyase digestion. Significant changes were identified predominantly in the case of chondroitin sulfate; for example, the total amount was higher in tumor tissue compared to the adjacent normal tissue. We also observed differences in the degree of sulfation and relative proportions of individual chondroitin sulfate disaccharides between lung cancer types and adjacent normal tissue. Furthermore, the differences in the 6-O-/4-O-sulfation ratio of chondroitin sulfate were different between the lung cancer types. Our pilot study revealed that further investigation of the role of chondroitin sulfate chains and enzymes involved in their biosynthesis is an important aspect of lung cancer research.

Keywords:  HPLC; adenocarcinoma; chondroitin sulfate; glycosaminoglycan; heparan sulfate; large cell carcinoma; lung cancer; mass spectrometry; small cell carcinoma; squamous cell carcinoma

DOI:  https://doi.org/10.3390/ijms24087050
JACS Au. 2023 Apr 24. 3(4): 1185-1195

Well-Defined Heparin Mimetics Can Inhibit Binding of the Trimeric Spike of SARS-CoV-2 in a Length-Dependent Manner.

Lifeng Sun, Pradeep Chopra, Ilhan Tomris, Roosmarijn van der Woude, Lin Liu, Robert P de Vries, Geert-Jan Boons.

The emergence of new SARS-CoV-2 variants and the dangers of long-covid necessitate the development of broad-acting therapeutics that can reduce viral burden. SARS-CoV-2 employs heparan sulfate (HS) as an initial cellular attachment factor, and therefore, there is interest in developing heparin as a therapeutic for SARS-CoV-2. Its use is, however, complicated by structural heterogeneity and the risk of causing bleeding and thrombocytopenia. Here, we describe the preparation of well-defined heparin mimetics by a controlled head-to-tail assembly of HS oligosaccharides having an alkyne or azide moiety by copper-catalyzed azide-alkyne cycloaddition (CuAAC). Alkyne- and azide-containing sulfated oligosaccharides were prepared from a common precursor by modifying an anomeric linker with 4-pentynoic acid and by enzymatic extension with an N-acetyl-glucosamine having an azide moiety at C-6 (GlcNAc6N3), respectively, followed by CuAAC. The process of enzymatic extension with GlcNAc6N3 followed by CuAAC with the desired alkyne-containing oligosaccharides could be repeated to give compounds composed of 20 and 27 monosaccharides, respectively. The heparin mimetics could inhibit the binding of the SARS-CoV-2 spike or RBD to immobilized heparin or to Vero E6 cells. The inhibitory potency increased with increasing chain length, and a compound composed of four sulfated hexasaccharides linked by triazoles had a similar potency as unfractionated heparin. Sequence analysis and HS microarray binding studies with a wide range of RBDs of variants of concern indicate that they have maintained HS-binding capabilities and selectivities. The heparin mimetics exhibit no or reduced binding to antithrombin-III and platelet factor 4, respectively, which are associated with side effects.

DOI: https://doi.org/10.1021/jacsau.3c00042
Biomacromolecules. 2023 Apr 28.

Multi-step Strategies Toward Regioselectively Sulfated M-Rich Alginates.

Fabiana Esposito, Antonio Laezza, Valentina Gargiulo, Serena Traboni, Alfonso Iadonisi, Annalisa La Gatta, Chiara Schiraldi, Emiliano Bedini.

Sulfated alginates (ASs), as well as several artificially sulfated polysaccharides, show interesting bioactivities. The key factors for structure-activity relationships studies are the degree of sulfation and the distribution of the sulfate groups along the polysaccharide backbone (sulfation pattern). The former parameter can often be controlled through stoichiometry, while the latter requires the development of suitable chemical or enzymatic, regioselective methods and is still missing for ASs. In this work, a study on the regioselective installation of several different protecting groups on a d-mannuronic acid enriched (M-rich) alginate is reported in order to develop a semi-synthetic access to regioselectively sulfated AS derivatives. A detailed structural characterization of the obtained ASs revealed that the regioselective sulfation could be achieved complementarily at the O-2 or O-3 positions of M units through multi-step sequences relying upon a silylating or benzoylating reagent for the regioselective protection of M-rich alginic acid, followed by sulfation and deprotection.

DOI: https://doi.org/10.1021/acs.biomac.3c00045
Biol Pharm Bull. 2023 Apr 22.

Efficient endocytosis of the human lactoferrin N-lobe enhances its antiproliferative activity against human cancer cells.

Masao Nakamura, Akira Shiga, Ami Iimori, Takumi Matsuzaki.

  Human lactoferrin (hLF) is a glycosylated globular iron-binding protein with high functional versatility that elicits anticancer, neuroprotective, and anti-inflammatory effects. Some of the diverse functions of hLF are induced after its internalization into various cells via cell surface endocytosis receptors, such as proteoglycans, which contain glycosaminoglycan (GAG) chains. We have previously demonstrated that an hLF derivative comprising the N-terminal half of hLF (referred to as the N-lobe) is internalized by intestinal enterocyte Caco-2 cells. However, the relationship between the intracellular uptake of the N-lobe and its pharmacological activity remains poorly understood. Here, we report that the N-lobe is efficiently internalized by lung cancer cells via endocytic pathways, suppressing their proliferation. Moreover, the N-lobe showed higher intracellular uptake than hLF. We found that the N-lobe was internalized into the human lung cancer cell lines PC-14 and PC-3 via clathrin- and/or caveolae-mediated endocytosis. Intracellular uptake of the N-lobe was inhibited when an equimolar concentration of chondroitin sulfate (CS)-E, a GAG subtype involved in malignant transformation and tumor metastasis, was added. The inhibitory effect of the N-lobe on PC-14 cell proliferation decreased with the addition of CS-E in a dose-dependent manner, suggesting that the CS-recognizing sequence on the N-lobe is necessary for its internalization or that the CS proteoglycan on cancer cells acts as an endocytosis receptor. These results suggest that the efficient endocytic uptake of the N-lobe is important for its antiproliferation effects on lung cancer cell lines. Thus, the N-lobe presents a promising drug candidate for cancer treatment.

Keywords:  Lactoferrin N-lobe; anticancer drug; cancer cell proliferative inhibition; chondroitin sulfate; endocytosis

DOI:  https://doi.org/10.1248/bpb.b23-00011
Mar Drugs. 2023 Mar 30. pii: 221. [Epub ahead of print]21(4):

Structural Characterization and Effects on Insulin Resistance of a Novel Chondroitin Sulfate from Halaelurus burgeri Skin.

Shiwei Hu, Hongli Zhu, Sichun Chen, Xiaofeng Wan, Yishu Liu, Zhaocai Ren, Shuang Gao.

  Several studies have isolated chondroitin sulphate (CHS) from sharks' jaws or cartilage. However, there has been little research on CHS from shark skin. In the present study, we extracted a novel CHS from Halaelurus burgeri skin, which has a novel chemical structure and bioactivity on improvement in insulin resistance. Results using Fourier transform-infrared spectroscopy (FT-IR), 1H-nuclear magnetic resonance spectroscopy (1H-NMR), and methylation analysis showed that the structure of the CHS was [4)-β-D-GlcpA-(1→3)-β-D-GlcpNAc-(1→]n with 17.40% of sulfate group concentration. Its molecular weight was 238.35 kDa, and the yield was 17.81%. Experiments on animals showed that this CHS could dramatically decrease body weight, reduce blood glucose and insulin levels, lower lipid concentrations both in the serum and the liver, improve glucose tolerance and insulin sensitivity, and regulate serum-inflammatory factors. These results demonstrated that the CHS from H. burgeri skin has a positive effect in reducing insulin resistance because of its novel structure, which provides a significant implication for the polysaccharide as a functional food.

Keywords:  Halaelurus burgeri skin; chemical structure; chondroitin sulphate; insulin resistance

DOI:  https://doi.org/10.3390/md21040221
J Exp Clin Cancer Res. 2023 Apr 28. 42(1): 106

Bispecific T cell-engager targeting oncofetal chondroitin sulfate induces complete tumor regression and protective immune memory in mice.

Nanna Skeltved, Mie A Nordmaj, Nicolai T Berendtsen, Robert Dagil, Emilie M R Stormer, Nader Al-Nakouzi, Ke Jiang, Alexandra Aicher, Christopher Heeschen, Tobias Gustavsson, Swati Choudhary, Ismail Gögenur, Jan P Christensen, Thor G Theander, Mads Daugaard, Ali Salanti, Morten A Nielsen.

   BACKGROUND: The malaria protein VAR2CSA binds oncofetal chondroitin sulfate (ofCS), a unique chondroitin sulfate, expressed on almost all mammalian cancer cells. Previously, we produced a bispecific construct targeting ofCS and human T cells based on VAR2CSA and anti-CD3 (V-aCD3Hu). V-aCD3Hu showed efficacy against xenografted tumors in immunocompromised mice injected with human immune cells at the tumor site. However, the complex effects potentially exerted by the immune system as a result of the treatment cannot occur in mice without an immune system. Here we investigate the efficacy of V-aCD3Mu as a monotherapy and combined with immune checkpoint inhibitors in mice with a fully functional immune system.
METHODS: We produced a bispecific construct consisting of a recombinant version of VAR2CSA coupled to an anti-murine CD3 single-chain variable fragment. Flow cytometry and ELISA were used to check cell binding capabilities and the therapeutic effect was evaluated in vitro in a killing assay. The in vivo efficacy of V-aCD3Mu was then investigated in mice with a functional immune system and established or primary syngeneic tumors in the immunologically "cold" 4T1 mammary carcinoma, B16-F10 malignant melanoma, the pancreatic KPC mouse model, and in the immunologically "hot" CT26 colon carcinoma model.
RESULTS: V-aCD3Mu had efficacy as a monotherapy, and the combined treatment of V-aCD3Mu and an immune checkpoint inhibitor showed enhanced effects resulting in the complete elimination of solid tumors in the 4T1, B16-F10, and CT26 models. This anti-tumor effect was abscopal and accompanied by a systemic increase in memory and activated cytotoxic and helper T cells. The combined treatment also led to a higher percentage of memory T cells in the tumor without an increase in regulatory T cells. In addition, we observed partial protection against re-challenge in a melanoma model and full protection in a breast cancer model.
CONCLUSIONS: Our findings suggest that V-aCD3Mu combined with an immune checkpoint inhibitor renders immunologically "cold" tumors "hot" and results in tumor elimination. Taken together, these data provide proof of concept for the further clinical development of V-aCD3 as a broad cancer therapy in combination with an immune checkpoint inhibitor.

Keywords:  Bispecific antibodies; Cancer; Checkpoint inhibitor; Immunotherapy; T cell memory; T cells therapy; Targeted therapy; VAR2CSA

DOI:  https://doi.org/10.1186/s13046-023-02655-8
Pharmaceutics. 2023 Apr 06. pii: 1163. [Epub ahead of print]15(4):

In Vitro Evaluation of Aerosol Therapy with Pentamidine-Loaded Liposomes Coated with Chondroitin Sulfate or Heparin for the Treatment of Leishmaniasis.

Lucía Román-Álamo, Mohamad Allaw, Yunuen Avalos-Padilla, Maria Letizia Manca, Maria Manconi, Federica Fulgheri, Jorge Fernández-Lajo, Luis Rivas, José Antonio Vázquez, José Esteban Peris, Xavier Roca-Geronès, Srisupaph Poonlaphdecha, Maria Magdalena Alcover, Roser Fisa, Cristina Riera, Xavier Fernàndez-Busquets.

  The second-line antileishmanial compound pentamidine is administered intramuscularly or, preferably, by intravenous infusion, with its use limited by severe adverse effects, including diabetes, severe hypoglycemia, myocarditis and renal toxicity. We sought to test the potential of phospholipid vesicles to improve the patient compliance and efficacy of this drug for the treatment of leishmaniasis by means of aerosol therapy. The targeting to macrophages of pentamidine-loaded liposomes coated with chondroitin sulfate or heparin increased about twofold (up to ca. 90%) relative to noncoated liposomes. The encapsulation of pentamidine in liposomes ameliorated its activity on the amastigote and promastigote forms of Leishmania infantum and Leishmania pifanoi, and it significantly reduced cytotoxicity on human umbilical endothelial cells, for which the concentration inhibiting 50% of cell viability was 144.2 ± 12.7 µM for pentamidine-containing heparin-coated liposomes vs. 59.3 ± 4.9 µM for free pentamidine. The deposition of liposome dispersions after nebulization was evaluated with the Next Generation Impactor, which mimics human airways. Approximately 53% of total initial pentamidine in solution reached the deeper stages of the impactor, with a median aerodynamic diameter of ~2.8 µm, supporting a partial deposition on the lung alveoli. Upon loading pentamidine in phospholipid vesicles, its deposition in the deeper stages significantly increased up to ~68%, and the median aerodynamic diameter decreased to a range between 1.4 and 1.8 µm, suggesting a better aptitude to reach the deeper lung airways in higher amounts. In all, nebulization of liposome-encapsulated pentamidine improved the bioavailability of this neglected drug by a patient-friendly delivery route amenable to self-administration, paving the way for the treatment of leishmaniasis and other infections where pentamidine is active.

Keywords:  Leishmania infantum; Leishmania pifanoi; aerosol therapy; drug encapsulation; leishmaniasis; liposomes; pentamidine

DOI:  https://doi.org/10.3390/pharmaceutics15041163
Int J Biol Macromol. 2023 Apr 24. pii: S0141-8130(23)01453-8. [Epub ahead of print] 124559

A branched 2-O sulfated 1,3-/1,4-galactoglucan from Antrodia cinnamomea exhibits moderate antiproliferative and anti-inflammatory activities.

Mei-Kuang Lu, Chi-Hsein Chao, Tsu-Yuan Chang, Ming-Che Cheng, Yu-Chi Hsu, Chia-Chuan Chang.

  A sulfated galactoglucan (3-SS) was discovered in Antrodia cinnamomea with antiproliferative and anti-inflammatory activities. Chemical identification of 3-SS resulted in the determination of a partial repeat unit as a 2-O sulfated 1,3-/1,4-linked galactoglucan with a two-residual 1,6-O-β-Glc branch on the 3-O position of a Glc. by monosaccharide analysis and 1D and 2D NMR spectroscopy. The anti-inflammation effects of 3-SS on RAW264.7 macrophage cells, such as IL-6 inhibition, restoration of LPS-induced IκB protein degradation, and inhibited LPS-induced TGFRII protein degradation, were confirmed to occur via AKT, ERK1/2, and p-38. In addition, 3-SS impaired the proliferation of H1975 lung cancer cells through EGFR/ERK/slug signaling. This is the first finding of 2-O sulfated 1,3-/1,4-galactoglucan with 1,6-β-Glc branches with dual functions of anti-inflammatory and antiproliferative activities.

Keywords:  Anti-inflammatory effects; Antiproliferation; Antrodia cinnamomea; Sulfated polysaccharides

DOI:  https://doi.org/10.1016/j.ijbiomac.2023.124559
Mar Drugs. 2023 Mar 24. pii: 205. [Epub ahead of print]21(4):

Glycosaminoglycans from the Starfish Lethasterias fusca: Structures and Influence on Hematopoiesis.

Maria I Bilan, Natalia Yu Anisimova, Alexandra I Tokatly, Sofya P Nikogosova, Dmitriy Z Vinnitskiy, Nadezhda E Ustyuzhanina, Andrey S Dmitrenok, Evgenia A Tsvetkova, Mikhail V Kiselevskiy, Nikolay E Nifantiev, Anatolii I Usov.

  Crude anionic polysaccharides extracted from the Pacific starfish Lethasterias fusca were purified by anion-exchange chromatography. The main fraction LF, having MW 14.5 kDa and dispersity 1.28 (data of gel-permeation chromatography), was solvolytically desulfated and giving rise to preparation LF-deS with a structure of dermatan core [→3)-β-d-GalNAc-(1→4)-α-l-IdoA-(1→]n, which was identified according to NMR spectroscopy data. Analysis of the NMR spectra of the parent fraction LF led to identification of the main component as dermatan sulfate LF-Derm →3)-β-d-GalNAc4R-(1→4)-α-l-IdoA2R3S-(1→ (where R was SO3 or H), bearing sulfate groups at O-3 or both at O-2 and O-3 of α-l-iduronic acid, as well as at O-4 of some N-acetyl-d-galactosamine residues. The minor signals in NMR spectra of LF were assigned as resonances of heparinoid LF-Hep composed of the fragments →4)-α-d-GlcNS3S6S-(1→4)-α-l-IdoA2S3S-(1→. The 3-O-sulfated and 2,3-di-O-sulfated iduronic acid residues are very unusual for natural glycosaminoglycans, and further studies are needed to elucidate their possible specific influence on the biological activity of the corresponding polysaccharides. To confirm the presence of these units in LF-Derm and LF-Hep, a series of variously sulfated model 3-aminopropyl iduronosides were synthesized and their NMR spectra were compared with those of the polysaccharides. Preparations LF and LF-deS were studied as stimulators of hematopoiesis in vitro. Surprisingly, it was found that both preparations were active in these tests, and hence, the high level of sulfation is not necessary for hematopoiesis stimulation in this particular case.

Keywords:  Lethasterias fusca; dermatan sulfate; hematopoiesis; heparinoid; starfish; synthetic sulfated iduronosides

DOI:  https://doi.org/10.3390/md21040205
Toxins (Basel). 2023 Mar 25. pii: 242. [Epub ahead of print]15(4):

The Uremic Toxin Indoxyl Sulfate Accelerates Senescence in Kidney Proximal Tubule Cells.

Yi Yang, Milos Mihajlovic, Manoe J Janssen, Rosalinde Masereeuw.

  Kidney fibrosis is the common final pathway of nearly all chronic and progressive nephropathies. One cause may be the accumulation of senescent cells that secrete factors (senescence associated secretory phenotype, SASP) promoting fibrosis and inflammation. It has been suggested that uremic toxins, such as indoxyl sulfate (IS), play a role in this. Here, we investigated whether IS accelerates senescence in conditionally immortalized proximal tubule epithelial cells overexpressing the organic anion transporter 1 (ciPTEC-OAT1), thereby promoting kidney fibrosis. Cell viability results suggested that the tolerance of ciPTEC-OAT1 against IS increased in a time-dependent manner at the same dose of IS. This was accompanied by SA-β-gal staining, confirming the accumulation of senescent cells, as well as an upregulation of p21 and downregulation of laminB1 at different time points, accompanied by an upregulation in the SASP factors IL-1β, IL-6 and IL-8. RNA-sequencing and transcriptome analysis revealed that IS accelerates senescence, and that cell cycle appears to be the most relevant factor during the process. IS accelerates senescence via TNF-α and NF-ĸB signalling early on, and the epithelial-mesenchymal transition process at later time points. In conclusion, our results suggest that IS accelerates cellular senescence in proximal tubule epithelial cells.

Keywords:  conditionally immortalized proximal tubule epithelial cells; kidney fibrosis; senescence-associated secretory phenotype (SASP); transcriptome

DOI:  https://doi.org/10.3390/toxins15040242
Int J Mol Sci. 2023 Apr 18. pii: 7452. [Epub ahead of print]24(8):

Detailing Protein-Bound Uremic Toxin Interaction Mechanisms with Human Serum Albumin in the Pursuit of Designing Competitive Binders.

Vida Dehghan Niestanak, Larry D Unsworth.

  Chronic kidney disease is the gradual progression of kidney dysfunction and involves numerous co-morbidities, one of the leading causes of mortality. One of the primary complications of kidney dysfunction is the accumulation of toxins in the bloodstream, particularly protein-bound uremic toxins (PBUTs), which have a high affinity for plasma proteins. The buildup of PBUTs in the blood reduces the effectiveness of conventional treatments, such as hemodialysis. Moreover, PBUTs can bind to blood plasma proteins, such as human serum albumin, alter their conformational structure, block binding sites for other valuable endogenous or exogenous substances, and exacerbate the co-existing medical conditions associated with kidney disease. The inadequacy of hemodialysis in clearing PBUTs underscores the significance of researching the binding mechanisms of these toxins with blood proteins, with a critical analysis of the methods used to obtain this information. Here, we gathered the available data on the binding of indoxyl sulfate, p-cresyl sulfate, indole 3-acetic acid, hippuric acid, 3-carboxyl-4-methyl-5-propyl-2-furan propanoic acid, and phenylacetic acid to human serum albumin and reviewed the common techniques used to investigate the thermodynamics and structure of the PBUT-albumin interaction. These findings can be critical in investigating molecules that can displace toxins on HSA and improve their clearance by standard dialysis or designing adsorbents with greater affinity for PBUTs than HSA.

Keywords:  binding affinity; chronic kidney disease; human serum albumin; protein-bound uremic toxins

DOI:  https://doi.org/10.3390/ijms24087452
bioRxiv. 2023 Apr 13. pii: 2023.04.12.536614. [Epub ahead of print]

Predictable genetic recruitment for luciferin sulfation in the convergent evolution of bioluminescence.

Emily S Lau, Jessica A Goodheart, Nolan T Anderson, Vannie L Liu, Arnab Mukherjee, Todd H Oakley.

Determinism and contingency shape evolutionary trajectories and the extent to which evolution is predictable depends on the relative contribution of each. However, the causes of patterns of predictable evolution are poorly understood. Here we propose evolution may be more deterministic - and therefore predictable - if functional evolution is constrained to fewer genetic solutions. We examine the bioluminescence system of ostracods and synthesize our findings with those reported previously for fireflies and sea pansies to show one aspect of bioluminescence, substrate metabolism, evolved convergently by recruiting homologous enzymes. We use gene expression, recombinant protein expression, and in vitro functional analyses to identify genes capable of substrate metabolism in ostracods. Our results, taken together with knowledge of firefly and sea pansy bioluminescence systems, provide empirical evidence supporting that these convergent mechanisms for substrate metabolism repeatedly evolved by co-opting sulfotransferases, a ubiquitous family of enzymes with broad substrate specificity. From this synthesis, we propose the lack of diverse genetic solutions for sulfation constrains evolution, resulting in more evolutionary determinism and increasing our ability to predict evolution.

DOI: https://doi.org/10.1101/2023.04.12.536614
Molecules. 2023 Apr 07. pii: 3297. [Epub ahead of print]28(8):

Between the Devil and the Deep Blue Sea-Resveratrol, Sulfotransferases and Sulfatases-A Long and Turbulent Journey from Intestinal Absorption to Target Cells.

Izabela Szymkowiak, Malgorzata Kucinska, Marek Murias.

  For nearly 30 years, resveratrol has attracted the scientific community's interest. This has happened thanks to the so-called French paradox, that is, the paradoxically low mortality from cardiovascular causes in the French population despite a diet rich in saturated fat. This phenomenon has been linked to the consumption of red wine, which contains a relatively high level of resveratrol. Currently, resveratrol is valued for its versatile, beneficial properties. Apart from its anti-atherosclerotic activity, resveratrol's antioxidant and antitumor properties deserve attention. It was shown that resveratrol inhibits tumour growth at all three stages: initiation, promotion, and progression. Moreover, resveratrol delays the ageing process and has anti-inflammatory, antiviral, antibacterial, and phytoestrogenic properties. These favorable biological properties have been demonstrated in vitro and in vivo in animal and human models. Since the beginning of the research on resveratrol, its low bioavailability, mainly due to its rapid metabolism, especially the first-pass effect that leaves almost no free resveratrol in the peripheral circulation, has been indicated as a drawback that has hindered its use. The elucidation of such issues as pharmacokinetics, stability, and the biological activity of resveratrol metabolites is therefore crucial for understanding the biological activity of resveratrol. Second-phase metabolism enzymes are mainly involved in RSV metabolism, e.g., UDP-glucuronyl transferases and sulfotransferases. In the present paper, we took a closer look at the available data on the activity of resveratrol sulfate metabolites and the role of sulfatases in releasing active resveratrol in target cells.

Keywords:  CYP450; resveratrol; resveratrol sulfate; sulfatase; sulfotransferase

DOI:  https://doi.org/10.3390/molecules28083297
Pharmaceutics. 2023 Mar 31. pii: 1115. [Epub ahead of print]15(4):

Approaches to Assure Similarity between Pharmaceutical Heparins from Two Different Manufacturers.

Francisco Felipe Bezerra, Stephan N M C G Oliveira, Rodrigo A Sales, Adriana A Piquet, Nina V Capillé, Eduardo Vilanova, Ana M F Tovar, Paulo A S Mourão.

  Pharmaceutical heparins from different manufacturers may present heterogeneities due to particular extraction and purification procedures or even variations in the raw material manipulation. Heparins obtained from different tissues also differ in their structure and activity. Nevertheless, there is an increased demand for more accurate assessments to ensure the similarities of pharmaceutical heparins. We propose an approach to accurately assess the similarity of these pharmaceutical preparations based on well-defined criteria, which are verified with a variety of refined analytical methods. We evaluate six commercial batches from two different manufacturers which were formulated with Brazilian or Chinese active pharmaceutical ingredients. Biochemical and spectroscopic methods and analysis based on digestion with heparinases were employed to evaluate the purity and structure of the heparins. Specific assays were employed to evaluate the biological activity. We observed minor but significant differences between the constitutive units of the heparins from these two manufacturers, such as the content of N-acetylated α-glucosamine. They also have minor differences in their molecular masses. These physicochemical differences have no impact on the anticoagulant activity but can indicate particularities on their manufacturing processes. The protocol we propose here for analyzing the similarity of unfractionated heparins is analogous to those successfully employed to compare low-molecular-weight heparins.

Keywords:  NMR analysis; anticoagulant; glycosaminoglycans; interchangeable drugs; structure of heparin

DOI:  https://doi.org/10.3390/pharmaceutics15041115
Sci Rep. 2023 Apr 28. 13(1): 6959

Sulfated endospermic nanocellulose crystals prevent the transmission of SARS-CoV-2 and HIV-1.

Enrique Javier Carvajal-Barriga, Wendy Fitzgerald, Emilios K Dimitriadis, Leonid Margolis, R Douglas Fields.

Biomaterials with antimicrobial activity are gaining attention due to their biodegradability and efficacy in interacting with a wide range of microorganisms. A new cellulose nano-biomaterial, endospermic nanocellulose crystals (ENC) obtained from parenchymal tissue of ivory nut endosperm, has a natural capacity as a universal binder. This feature is enhanced when it is chemically functionalized, and can be exploited in the fight against microbes. We tested the ability of sulfated ENC in aqueous suspension to encapsulate viruses through a crosslinking reaction mediated by cations. 0.25% w/v ENC suspensions efficiently encapsulated spike (S) protein, preventing its interaction with ACE2 receptor. ENC was further able to encapsulate SARS-CoV-2 pseudoviruses and prevent infection of 293T-hsACE2 cells. ENC also suppressed infection of MT-4 cells with HIV-1LAI.04. This antiviral activity of sulfated ENC is due to the irreversible interaction of ENC with viral particles mediated by crosslinking, as antiviral activity was less effective in the absence of cations. Additionally, ENC was used as a matrix to immobilize recombinant ACE2 receptors and anti-S IgG, creating molecular lures that efficiently inhibited SARS-CoV-2 infections in vitro. These results show that sulfated ENC from ivory nuts can be used as an efficient antiviral material.

DOI: https://doi.org/10.1038/s41598-023-33686-y
BMC Endocr Disord. 2023 Apr 23. 23(1): 89

Correlation between biochemical and clinical hyperandrogenism parameter in polycystic ovary syndrome in relation to age.

Zaixin Guo, Fengjun Jin, Shuwen Chen, Pan Hu, Yanfang Hao, Qi Yu.

   BACKGROUND: To assess the correlation between clinical and biochemical hyperandrogenism parameters in polycystic ovary syndrome (PCOS) according to age.
METHODS: This prospective study included 256 PCOS patients diagnosed according to the Rotterdam criteria in a university-based hospital. Androgen levels were measured using liquid chromatography-tandem mass spectrometry. Hirsutism, acne, and alopecia were assessed using the modified Ferriman-Gallwey (mF-G) score, Comprehensive Acne Severity Scale (CASS), and the Ludwig scale, respectively. The correlation between biochemical and clinical hyperandrogenism parameters was assessed in younger and older women with PCOS.
RESULTS: The 256 PCOS patients were classified by age into two groups: age 18-29 years (n = 151) and age 30-40 years (n = 84). In women with PCOS, mF-G was significantly positively correlated with the free androgen index (FAI), dehydroepiandrosterone (DHEA), and DHEA sulfate (DHEA-S). CASS had a significant positive correlation with DHEA. mF-G was positively correlated with FAI in those aged 18-29 years, but the correlations were not significant in those aged 30-40 years. The positive correlation between specific body regions of clinical hyperandrogenism, especially mF-G of chin, lower abdomen, and thighs, and testosterone, as well as with FAI, was highest in those aged 18-29 years. In those aged 30-40 years clinical hyperandrogenism was mainly affected by DHEA, DHEA-S, and dihydrotestosterone.
CONCLUSION: The correlation between biochemical and clinical hyperandrogenism parameters varied with age in our East Asian population. Clinical hyperandrogenism was positively correlated with FAI in younger women with PCOS. The correlation between biochemical and clinical hyperandrogenism was not significant in older women with PCOS.

Keywords:  Age; Dehydroepiandrosterone; Hyperandrogenism; Polycystic ovary syndrome

DOI:  https://doi.org/10.1186/s12902-023-01346-x
Food Res Int. 2023 05;pii: S0963-9969(23)00178-3. [Epub ahead of print]167 112633

Metabolites of sea cucumber sulfated polysaccharides fermented by Parabacteroides distasonis and their effects on cross-feeding.

Zhengqi Liu, Yuanyuan Hu, Xiaoya Tao, Jinjin Li, Xiaoming Guo, Gang Liu, Shuang Song, Beiwei Zhu.

  Sea cucumber sulfated polysaccharide (SCSPsj) is one of the dietary components which effectively modulates gut microbiota; however, the underlying mechanism remains unclear. In the present study, the interaction between SCSPsj and its utilizer (Parabacteroides distasonis) was investigated. Further study was carried out to explore the cross-feeding between intestinal Bacteroidales mediated by SCSPsj. The results revealed that SCSPsj can be fermented by P. distasonis to produce various microbial metabolites, including organic acids and derivatives, lipids and lipid-like molecules, organoheterocyclic compounds. SCSPsj can regulate the succinate pathway and acetyl-CoA pathway to influence the production of propanoic acid and acetic acid, respectively. Moreover, the SCSPsj-fermented supernatants of P. distasonis can only promote the growth of B. stercoris, B. vulgatus and P. johnsonii among 8 intestinal Bacteroidales strains through cross-feeding. The effect of cross-feeding was related to spatial distances and bacterial species. Moreover, the cross-feeding was correlated with compounds belonging to organic acids and derivatives, lipids and lipid-like molecules. These findings could provide new insights into the interaction between SCSPsj and gut microbiota.

Keywords:  Cross-feeding; Metabolites; Parabacteroides distasonis; Sea cucumber sulfated polysaccharide

DOI:  https://doi.org/10.1016/j.foodres.2023.112633
Toxins (Basel). 2023 Apr 07. pii: 276. [Epub ahead of print]15(4):

Proton-Pump Inhibitors and Serum Concentrations of Uremic Toxins in Patients with Chronic Kidney Disease.

Chronic Kidney Disease-Renal Epidemiology and Information Network (CKD-REIN) Study Group

  Use of proton-pump inhibitors (PPIs) is common in patients with chronic kidney disease (CKD). PPIs and many uremic toxins (UTs) are eliminated by the kidney's tubular organic anion transporter system. In a cross-sectional study, we sought to evaluate the association between PPI prescription and serum concentrations of various UTs. We studied a randomly selected sub-group of participants in the CKD-REIN cohort (adult patients with a confirmed diagnosis of CKD and estimated glomerular filtration rate (eGFR) < 60 mL/min/1.73 m2) with available frozen samples collected at baseline. PPI prescription was recorded at baseline. Serum concentrations of 10 UTs were measured using a validated liquid chromatography tandem mass spectrometry technique. Multiple linear regression was performed, with the log UT concentration as the dependent variable. Of the 680 included patients (median age: 68 years; median eGFR: 32 mL/min/1.73 m2), 31% had PPI prescriptions at baseline. Patients using PPIs had higher levels of certain UTs in comparison to other patients, including total and free indoxyl sulfate (IS), total and free p-cresylsulfate, total and free p-cresylglucuronide (PCG), phenylacetylglutamine (PAG), free kynurenine, and free hippuric acid. After adjustment for baseline co-morbidities, number of co-prescribed drugs, and laboratory data, including eGFR, associations between PPI prescription and elevated serum concentrations of free and total IS, free and total PCG, and PAG remained significant. Our results indicate that PPI prescription is independently associated with serum UT retention. These findings are interesting to better understand the factors that may modulate serum UT concentration in CKD patients, however, they will need to be confirmed by longitudinal studies.

Keywords:  chronic kidney disease; indoxyl sulfate; proton-pump inhibitor; uremic toxins

DOI:  https://doi.org/10.3390/toxins15040276