bims-supasi 2022-12-11 papers

bims-supasi

Biomed News

on Sulfation pathways and signalling

Issue of 2022‒12‒11
ten papers selected by
Jonathan Wolf Mueller
University of Birmingham

Biochemical and crystallographic assessments of the effect of 4,6-O-disulfated disaccharide moieties in chondroitin sulfate E on chondroitinase ABC I activity.
Evaluation of unsulfated biotechnological chondroitin in a knee osteoarthritis mouse model as a potential novel functional ingredient in nutraceuticals and pharmaceuticals.
Extended-release of doxorubicin through green surface modification of gold nanoparticles: in vitro and in ovo assessment.
Preparation and properties of fucoxanthin-loaded liposomes stabilized by sea cucumber derived cholesterol sulfate instead of cholesterol.
Interaction of luteolin, naringenin, and their sulfate and glucuronide conjugates with human serum albumin, cytochrome P450 (CYP2C9, CYP2C19, and CYP3A4) enzymes and organic anion transporting polypeptide (OATP1B1 and OATP2B1) transporters.
Rapid and sustainable HPLC method for the determination of uremic toxins in human plasma samples.
Heparin Does Not Regulate Circulating Human PCSK9 (Proprotein Convertase Subtilisin-Kexin Type 9) in a General Population.
Novel insights into bile acid detoxification via CYP, UGT and SULT enzymes.
In vitro fermentation of heparin by the human gut microbiota: Changes in the microbiota community and metabolic functions.
Dehydroepiandrosterone sulphate and mortality in middle-aged and older men and women - a J-shaped relationship.

FEBS J. 2022 Nov 22.

Biochemical and crystallographic assessments of the effect of 4,6-O-disulfated disaccharide moieties in chondroitin sulfate E on chondroitinase ABC I activity.

Ippei Watanabe, Akimasa Miyanaga, Hirotaka Hoshi, Kiyoshi Suzuki, Tadashi Eguchi.

  O-sulfated N-acetyl-d-galactosamine (GalNAc) residues in chondroitin sulfate (CS) play a crucial role in chondroitinase ABC I (cABC-I) activity. CSA containing mainly 4-O-monosulfated GalNAc was a good substrate for the enzyme, but not CSE containing mainly 4,6-O-disulfated GalNAc [GalNAc(4S,6S)]. Each CS isomer exhibits structural heterogeneity; CSE has di-sulfated disaccharide units and mono-sulfated disaccharide units. Disaccharide composition analysis of digested products revealed that mono-sulfated disaccharide units in CSE contributed to the enzyme reactivity. Although enough substrate (CSA) was present in mixtures of CSA and CSE for reaction, the reactivity was reduced depending on the amount of CSE in the mixture. These results suggested that CSE is not only resistant to enzyme digestion but also attenuates enzyme activity. To understand the mechanism of action, crystallography of cABC-I in complex with unsaturated CSE-disaccharide, ΔDi-(4,6)S, was performed. Both 4-O- and 6-O-sulfate groups in ΔDi-(4,6)S interact with Arg500, suggesting that there was a greater interaction between ΔDi-(4,6)S and Arg500 than between mono-sulfated disaccharides and Arg500. Besides, this interaction attenuated enzyme activity by interfering with a function of Arg500, which is the charge neutralization of the carboxy group of D-glucuronic acid (GlcA) residues in CS. When interacting with the CSE-disaccharide unit [GlcAβ1-3GalNAc(4S,6S)] in CS, cABC-I cannot interact with other CS-disaccharide units until it has digested the CSE-disaccharide unit. The low reactivity of cABC-I with CSE is attributable to two suggested factors: (a) resistance of E-units in CSE molecules to digestion by cABC-I, and (b) tendency of E-units in CSE molecules to attenuate cABC-I activity.

Keywords:  4,6-O-disulfated; chondroitin sulfate E; condoliase; crystal structure; substrate reactivity

DOI:  https://doi.org/10.1111/febs.16685
Front Bioeng Biotechnol. 2022 ;10 934997

Evaluation of unsulfated biotechnological chondroitin in a knee osteoarthritis mouse model as a potential novel functional ingredient in nutraceuticals and pharmaceuticals.

Donatella Cimini, Serena Boccella, Alberto Alfano, Antonietta Stellavato, Salvatore Paino, Chiara Schiraldi, Francesca Guida, Michela Perrone, Maria Donniacuo, Virginia Tirino, Vincenzo Desiderio, Barbara Rinaldi.

  Osteoarthritis is a very disabling disease that can be treated with both non-pharmacological and pharmacological approaches. In the last years, pharmaceutical-grade chondroitin sulfate (CS) and glucosamine emerged as symptomatic slow-acting molecules, effective in pain reduction and improved function in patients affected by osteoarthritis. CS is a sulfated glycosaminoglycan that is currently produced mainly by extraction from animal tissues, and it is commercialized as a pharmaceutical-grade ingredient and/or food supplement. However, public concern on animal product derivatives has prompted the search for alternative non-extractive production routes. Thus, different approaches were established to obtain animal-free natural identical CS. On the other hand, the unsulfated chondroitin, which can be obtained via biotechnological processes, demonstrated promising anti-inflammatory properties in vitro, in chondrocytes isolated from osteoarthritic patients. Therefore, the aim of this study was to explore the potential of chondroitin, with respect to the better-known CS, in an in vivo mouse model of knee osteoarthritis. Results indicate that the treatment with biotechnological chondroitin (BC), similarly to CS, significantly reduced the severity of mechanical allodynia in an MIA-induced osteoarthritic mouse model. Decreased cartilage damage and a reduction of inflammation- and pain-related biochemical markers were also observed. Overall, our data support a beneficial activity of biotechnological unsulfated chondroitin in the osteoarthritis model tested, thus suggesting BC as a potential functional ingredient in pharmaceuticals and nutraceuticals with the advantage of avoiding animal tissue extraction.

Keywords:  chondroitin; chondroitin sulfate; functional ingredient; mouse model; osteoarthritis

DOI:  https://doi.org/10.3389/fbioe.2022.934997
BMC Chem. 2022 Dec 06. 16(1): 110

Extended-release of doxorubicin through green surface modification of gold nanoparticles: in vitro and in ovo assessment.

Maryam Asariha, Seyed Hossein Kiaie, Sepideh Izadi, Faezeh H Pirhayati, Mehdi Fouladi, Maryam Gholamhosseinpour.

  In the present study, a green surface modification of gold nanoparticles (GNPs) using chondroitin sulfate (CHS) and chitosan (CS) to deliver an extended-release of doxorubicin (DOX) was proposed. Following synthesis of each step of unconjugated counterpart, including CHS-GNPs, DOX-CHS-GNP, and conjugated construct DOX-CHS-GNP-CS, physicochemical properties of the nanoparticles (NPs) were characterized by FT-IR, DLS, and TEM analyses, and the release of DOX was determined by using UV-Vis spectrometry. Then, NPs were effectively taken up by MDA-MB-468, βTC-3, and human fibroblast (HFb) cell lines with high release percent and without significant cytotoxicity. The DOX-CHS-GNPs and DOX-CHS-GNP-CS NPs showed a mean size of 175.8 ± 1.94 and 208.9 ± 2.08 nm; furthermore, a zeta potential of - 34 ± 5.6 and - 25.7 ± 5.9 mV, respectively. The highest release of DOX was 73.37% after 45 h, while in the absence of CS, the release of DOX was 76.05% for 24 h. Compared to CHS-GNPs, the presence of CS decreased the rate of sustained release of DOX and improved the drug release efficiency. The results demonstrated an excellent release and negligible cytotoxicity at high concentrations of CHS-GNP-CS. Consequently, in ovo assessment corroborated the efficacy of the green fabricated NPs proposed effective targeted delivery of DOX for anti-tumor therapy in vitro.

Keywords:  Chitosan; Chondroitin sulfate; Cytotoxicity; Doxorubicin; Drug delivery; Gold nanoparticles; Green synthesis

DOI:  https://doi.org/10.1186/s13065-022-00895-x
J Biosci Bioeng. 2022 Dec 06. pii: S1389-1723(22)00343-7. [Epub ahead of print]

Preparation and properties of fucoxanthin-loaded liposomes stabilized by sea cucumber derived cholesterol sulfate instead of cholesterol.

Wang Yunling, Yang Jinyue, Wang Yuming, Chang Yaoguang, Xue Changhu, Zhang Tiantian.

  The preparation of steady-state phospholipid liposomes requires cholesterol as a stabilizer, but excessive intake of cholesterol may increase the risk of cardiovascular disease. The sulfated sterols extracted from sea cucumber, mainly including sulfated 24-methylene cholesterol and cholesterol sulfate, have been reported to have a variety of physiological activities. Sulfated sterols are similar to cholesterol in structure and have the potential to replace cholesterol to prepare novel stable multifunctional liposomes, allowing the liposomes to act as carriers for the delivery of less bioavailable nutrients while allowing sulfated sterols in the lipid bilayer to exert physiologically active effects. This study aimed to prepare a novel multifunctional nanoliposome stabilized with sulfated sterols from sea cucumber instead of cholesterol by ultrasound-assisted thin-film dispersion method. The results showed that stable and uniformly dispersed nanoliposomes could be formed when the substitution ratio of sea cucumber-derived cholesterol sulfate was 100% and the ratio of lecithin to cholesterol sulfate was 3:1. Fucoxanthin encapsulated liposome with egg yolk lecithin/sea cucumber-derived cholesterol sulfate/fucoxanthin mass ratio of 6:2:3 was successfully prepared, with an average particle size of 214 ± 3 nm, polydispersity index (PDI) value of 0.297 ± 0.006, the zeta potential of -57.2 ± 1.10 mV, and the encapsulation efficiency of 85.5 ± 0.8%. The results of digestion and absorption in vitro and in vivo showed that liposomes could significantly improve the bioavailability of fucoxanthin and prolong its residence time in serum. As an efficient multifunctional carrier, this novel liposome has great potential for applications in functional foods and biomedicine.

Keywords:  Absorption; Cholesterol sulfate; Digestion; Fucoxanthin; Liposome; Sea cucumber sulfated sterols

DOI:  https://doi.org/10.1016/j.jbiosc.2022.11.004
Biomed Pharmacother. 2022 Dec 05. pii: S0753-3322(22)01467-6. [Epub ahead of print]157 114078

Interaction of luteolin, naringenin, and their sulfate and glucuronide conjugates with human serum albumin, cytochrome P450 (CYP2C9, CYP2C19, and CYP3A4) enzymes and organic anion transporting polypeptide (OATP1B1 and OATP2B1) transporters.

Hana Kaci, Slávka Bodnárová, Eszter Fliszár-Nyúl, Beáta Lemli, Helena Pelantová, Kateřina Valentová, Éva Bakos, Csilla Özvegy-Laczka, Miklós Poór.

  Luteolin and naringenin are flavonoids found in various foods/beverages and present in certain dietary supplements. After a high intake of these flavonoids, their sulfate and glucuronide conjugates reach micromolar concentrations in the bloodstream. Some pharmacokinetic interactions of luteolin and naringenin have been investigated in previous studies; however, only limited data are available in regard to their metabolites. In this study, we aimed to investigate the interactions of the sulfate and glucuronic acid conjugates of luteolin and naringenin with human serum albumin, cytochrome P450 (CYP2C9, 2C19, and 3A4) enzymes, and organic anion transporting polypeptide (OATP1B1 and OATP2B1) transporters. Our main findings are as follows: (1) Sulfate conjugates formed more stable complexes with albumin than the parent flavonoids. (2) Luteolin and naringenin conjugates showed no or only weak inhibitory action on the CYP enzymes examined. (3) Certain conjugates of luteolin and naringenin are potent inhibitors of OATP1B1 and/or OATP2B1 enzymes. (4) Conjugated metabolites of luteolin and naringenin may play an important role in the pharmacokinetic interactions of these flavonoids.

Keywords:  Cytochrome P450 enzymes; Human serum albumin; Luteolin; Naringenin; OATP transporters; Sulfate/glucuronide metabolites

DOI:  https://doi.org/10.1016/j.biopha.2022.114078
Anal Bioanal Chem. 2022 Dec 05.

Rapid and sustainable HPLC method for the determination of uremic toxins in human plasma samples.

Luís A P Silva, Stefano Campagnolo, Sara R Fernandes, Sara S Marques, Luisa Barreiros, Benedita Sampaio-Maia, Marcela A Segundo.

  Protein-bound uremic toxins, mainly indoxyl sulfate (3-INDS), p-cresol sulfate (pCS), and indole-3-acetic acid (3-IAA) but also phenol (Pol) and p-cresol (pC), are progressively accumulated during chronic kidney disease (CKD). Their accurate measurement in biomatrices is demanded for timely diagnosis and adoption of appropriate therapeutic measures. Multianalyte methods allowing the establishment of a uremic metabolite profile are still missing. Hence, the aim of this work was to develop a rapid and sensitive method based on high-performance liquid chromatography with fluorescence detection for the simultaneous quantification of Pol, 3-IAA, pC, 3-INDS, and pCS in human plasma. Separation was attained in 12 min, using a monolithic C18 column and isocratic elution with acetonitrile and phosphate buffer containing an ion-pairing reagent, at a flow rate of 2 mL min-1. Standards were prepared in plasma and quantification was performed using the background subtraction approach. LOQ values were ≤ 0.2 µg mL-1 for all analytes except for pCS (LOQ of 2 µg mL-1). The method proved to be accurate (93.5-112%) and precise (CV ≤ 14.3%). The multianalyte application of the method, associated to a reduced sample volume (50 µL), a less toxic internal standard (eugenol) in comparison to the previously applied 2,6-dimethylphenol and 4-ethylphenol, and a green extraction solvent (ethanol), resulted in the AGREE score of 0.62 which is in line with the recent trend of green and sustainable analytical chemistry. The validated method was successfully applied to the analysis of plasma samples from control subjects exhibiting normal levels of uremic toxins and CKD patients presenting significantly higher levels of 3-IAA, pC, 3-INDS, and pCS that can be further investigated as biomarkers of disease progression.

Keywords:  3-Indoxyl sulfate; Chronic kidney disease; Indole-3-acetic acid; Uremic toxin; p-Cresol; p-Cresol sulfate

DOI:  https://doi.org/10.1007/s00216-022-04458-w
Arterioscler Thromb Vasc Biol. 2022 Dec 08.

Heparin Does Not Regulate Circulating Human PCSK9 (Proprotein Convertase Subtilisin-Kexin Type 9) in a General Population.

Vivian Q Xia, Chui Mei Ong, Lucas S Zier, John S MacGregor, Alan H B Wu, John S Chorba.

  BACKGROUND: PCSK9 (proprotein convertase subtilisin-kexin type 9) chaperones the hepatic LDLR (low-density lipoprotein receptor) for lysosomal degradation, elevating serum LDL (low-density lipoprotein) cholesterol and promoting atherosclerotic heart disease. Though the major effect on the hepatic LDLR comes from secreted PCSK9, the details of PCSK9 reuptake into the hepatocyte remain unclear. In both tissue culture and animal models, HSPGs (heparan sulfate proteoglycans) on hepatocytes act as co-receptors to promote PCSK9 reuptake. We hypothesized that if this PCSK9:HSPG interaction is important in humans, disrupting it with unfractionated heparin (UFH) would acutely displace PCSK9 from the liver and increase plasma PCSK9.METHODS: We obtained remnant plasma samples from 160 subjects undergoing cardiac catheterization before and after administration of intravenous UFH. PCSK9 levels were determined using a commercial enzyme-linked immunosorbent assay.
RESULTS: Median plasma PCSK9 was 113 ng/mL prior to UFH and 119 ng/mL afterward. This difference was not significantly different (P=0.83 [95% CI, -6.23 to 6.31 ng/mL]). Equivalence testing provided 95% confidence that UFH would not raise plasma PCSK9 by > 4.7%. Among all subgroups, only subjects with the lowest baseline PCSK9 concentrations exhibited a response to UFH (8.8% increase, adj. P=0.044). A modest correlation was observed between baseline plasma PCSK9 and the change in plasma PCSK9 due to UFH (RS=-0.3634; P<0.0001).
CONCLUSIONS: Administration of UFH does not result in a clinically meaningful effect on circulating PCSK9 among an unselected population of humans. The results cast doubt on the clinical utility of disrupting the PCSK9:HSPG interaction as a general therapeutic strategy for PCSK9 inhibition. However, the observations suggest that in selected populations, disrupting the PCSK9:HSPG interaction could still affect PCSK9 reuptake and offer a therapeutic benefit.

Keywords:  animal models; cholesterol; heart disease; hepatocyte; liver

DOI:  https://doi.org/10.1161/ATVBAHA.122.318556
Toxicol In Vitro. 2022 Dec 03. pii: S0887-2333(22)00231-4. [Epub ahead of print] 105533

Novel insights into bile acid detoxification via CYP, UGT and SULT enzymes.

Vlasia Kastrinou Lampou, Birk Poller, Felix Huth, Audrey Fischer, Gerd A Kullak-Ublick, Michael Arand, Heiko S Schadt, Gian Camenisch.

  Bile acid (BA) homeostasis is a complex and precisely regulated process to prevent impaired BA flow and the development of cholestasis. Several reactions, namely hydroxylation, glucuronidation and sulfation are involved in BA detoxification. In the present study, we employed a comprehensive approach to identify the key enzymes involved in BA metabolism using human recombinant enzymes, human liver microsomes (HLM) and human liver cytosol (HLC). We showed that CYP3A4 was a crucial step for the metabolism of several BAs and their taurine and glycine conjugated forms and quantitatively described their metabolites. Glucuronidation and sulfation were also identified as important drivers of the BA detoxification process in humans. Moreover, lithocholic acid (LCA), the most hydrophobic BA with the highest toxicity potential, was a substrate for all investigated processes, demonstrating the importance of hepatic metabolism for its clearance. Collectively, this study identified CYP3A4, UGT1A3, UGT2B7 and SULT2A1 as the major contributing (metabolic) processes in the BA detoxification network. Inhibition of these enzymes by drug candidates is therefore considered as a critical mechanism in the manifestation of drug-induced cholestasis in humans and should be addressed during the pre-clinical development.

Keywords:  Bile acid metabolism; CYP3A4; Glucuronidation; LCA; Lithocholic acid; Sulfation

DOI:  https://doi.org/10.1016/j.tiv.2022.105533
Food Chem. 2022 Nov 21. pii: S0308-8146(22)02972-7. [Epub ahead of print]406 135010

In vitro fermentation of heparin by the human gut microbiota: Changes in the microbiota community and metabolic functions.

Chuan Zhang, Leilei Yu, Qixiao Zhai, Ruohan Zhao, Jianxin Zhao, Hao Zhang, Wei Chen, Fengwei Tian.

  Levels of its utilization suggest that the host glycosaminoglycan heparin is an important carbohydrate in the human gut microbiota. However, the interaction between heparin and the gut microbiota is not well understood. In this study, an in vitro fermentation system combined with microbiome and metabolome technologies was used to study the interaction between heparin and the gut microbiota. Interestingly, we found that heparin can be used by the gut microbiota, which produce large amounts of short chain fatty acids leading to a decrease in pH. In addition, the addition of heparin increased the relative abundance of Bacteroides and Bifidobacterium and decreased the relative abundance of Escherichia-Shigella. Correlation analysis of the microbiome and metabolome revealed that the catabolism of heparin was accompanied by the biosynthesis of bile acids and tryptophan metabolism. Overall, this study provides new evidence on the role of heparin as a stable carbon source for the gut microbiota and forms a strong basis for the use of heparin to condition the gut microbiota.

Keywords:  Fermentation; Gut microbiota; Heparin; Metabolome; Microbiome

DOI:  https://doi.org/10.1016/j.foodchem.2022.135010
J Clin Endocrinol Metab. 2022 Dec 08. pii: dgac716. [Epub ahead of print]

Dehydroepiandrosterone sulphate and mortality in middle-aged and older men and women - a J-shaped relationship.

Trasias Mukama, Theron Johnson, Verena Katzke, Rudolf Kaaks.

  CONTEXT: The relationship between DHEAS and mortality is of scientific and public health interest, yet remains poorly understood.OBJECTIVE: Examine the association between DHEAS and cancer, cardiovascular and all-cause mortality in middle-aged and older men and women.
DESIGN: Case-cohort nested within EPIC-Heidelberg. DHEAS was measured in 7,370 stored serum collected from 1994 to 1998. Median follow-up for incident mortality events: 17.7 years.
SETTING: General community.
PARTICIPANTS: The case-cohort included 7,370 men (mean age = 55.0) and women (mean age = 52.4 years). All deaths due to cancer (n = 1040), cardiovascular diseases (n = 598) and all causes (n = 2407) which occurred in EPIC-Heidelberg until end of 2014 were included.
RESULTS: The association between DHEAS and mortality was non-linear such that both participants in the lowest (Q1) and highest (Q5) sex- and 5-year age-group specific quintiles of DHEAS were at increased hazards of mortality from cardiovascular [Q1: HR = 1.83 95%CI: (1.33-2.51), Q5: 1.39 (1.00-1.94)], cancer [Q1: 1.27 (1.01-1.60), Q5: 1.27 (1.02-1.60)] and all causes [Q1: 1.51 (1.25-1.82), Q5: 1.31 (1.08-1.58)], compared to participants in Q3. In men and women with below median DHEAS levels, doubling of DHEAS was associated with lower hazards of cardiovascular [0.87, (0.78-0.96)], cancer [0.90, (0.83-0.97)], and total mortality [0.89, (0.83-0.95)]. In contrast, a doubling in DHEAS among participants with above median levels was associated with 1.20, (1.01-1.42), 1.28, (1.01-1.62) and 1.19 (1.03-1.37) higher hazards of mortality from cancer, cardiovascular, and all-causes respectively.
CONCLUSION: In this large population based study, DHEAS showed a J-shaped association with mortality. Both participants with lowest and highest levels experienced higher hazards of mortality from cancer, cardiovascular and all causes.

Keywords:  DHEAS; all-cause; cancer; cardiovascular disease; mortality

DOI:  https://doi.org/10.1210/clinem/dgac716