bims-supasi 2023-06-11 papers

bims-supasi

Biomed News

on Sulfation pathways and signalling

Issue of 2023‒06‒11
seventeen papers selected by
Jonathan Wolf Mueller
University of Birmingham

Chondroitin 4-O-sulfation regulates hippocampal perineuronal nets and social memory.
Biotechnological advances in the synthesis of modified chondroitin towards novel biomedical applications.
Chondroitin sulfate, dermatan sulfate, and hyaluronic acid differentially modify the biophysical properties of collagen-based hydrogels.
Rational Design and Expedient Synthesis of Heparan Sulfate Mimetics from Natural Aminoglycosides for Structure and Activity Relationship Studies.
Chondroitin Sulfate Alleviated Obesity by Modulating Gut Microbiota and Liver Metabolome in High-Fat-Diet-Induced Obese Mice.
Reduction of Erythema in Moderate-Severe Rosacea by a Low Molecular Weight Heparan Sulfate Analog (HSA).
Targeting heparan sulfate-protein interactions with oligosaccharides and monoclonal antibodies.
Infection-induced epilepsy is caused by increased expression of chondroitin sulfate proteoglycans in hippocampus and amygdala.
Assimilatory sulfate reduction in the marine methanogen Methanothermococcus thermolithotrophicus.
Sex Hormones associated with Temporomandibular Pain on Palpation in Male Adolescents - Results of the Epidemiologic LIFE Child Study.
Site-Specific Incorporation of Sulfotyrosine into Proteins in Mammalian Cells.
Association of size for gestational age and dehydroepiandrosterone sulfate with cardiometabolic risk in central precocious puberty girls.
Direct anti-tumoral effects of sulfated fucans isolated from echinoderms: a possible role of neuropilin-1/β1 integrin endocytosis and focal adhesion kinase (FAK) degradation.
Structural basis for FGF hormone signalling.
Circulating adrenal 11-oxygenated androgens are associated with clinical outcome in endometrial cancer.
Large-scale heparin-based bind-and-elute chromatography identifies two biologically distinct populations of extracellular vesicles.
Heparin Affinity-Based IL-4 Delivery to Modulate Macrophage Phenotype and Endothelial Cell Activity In Vitro.

Proc Natl Acad Sci U S A. 2023 Jun 13. 120(24): e2301312120

Chondroitin 4-O-sulfation regulates hippocampal perineuronal nets and social memory.

Huiqian Huang, Amélie M Joffrin, Yuan Zhao, Gregory M Miller, Grace C Zhang, Yuki Oka, Linda C Hsieh-Wilson.

  Glycan alterations are associated with aging, neuropsychiatric, and neurodegenerative diseases, although the contributions of specific glycan structures to emotion and cognitive functions remain largely unknown. Here, we used a combination of chemistry and neurobiology to show that 4-O-sulfated chondroitin sulfate (CS) polysaccharides are critical regulators of perineuronal nets (PNNs) and synapse development in the mouse hippocampus, thereby affecting anxiety and cognitive abilities such as social memory. Brain-specific deletion of CS 4-O-sulfation in mice increased PNN densities in the area CA2 (cornu ammonis 2), leading to imbalanced excitatory-to-inhibitory synaptic ratios, reduced CREB activation, elevated anxiety, and social memory dysfunction. The impairments in PNN densities, CREB activity, and social memory were recapitulated by selective ablation of CS 4-O-sulfation in the CA2 region during adulthood. Notably, enzymatic pruning of the excess PNNs reduced anxiety levels and restored social memory, while chemical manipulation of CS 4-O-sulfation levels reversibly modulated PNN densities surrounding hippocampal neurons and the balance of excitatory and inhibitory synapses. These findings reveal key roles for CS 4-O-sulfation in adult brain plasticity, social memory, and anxiety regulation, and they suggest that targeting CS 4-O-sulfation may represent a strategy to address neuropsychiatric and neurodegenerative diseases associated with social cognitive dysfunction.

Keywords:  chondroitin sulfate (CS); glycans; glycosaminoglycans (GAGs); perineuronal nets (PNNs); social memory

DOI:  https://doi.org/10.1073/pnas.2301312120
Biotechnol Adv. 2023 Jun 06. pii: S0734-9750(23)00092-7. [Epub ahead of print] 108185

Biotechnological advances in the synthesis of modified chondroitin towards novel biomedical applications.

Donatella Cimini, Emiliano Bedini, Chiara Schiraldi.

  Chondroitin sulfate (CS) is a well-known glycosaminoglycan present in a large variety of animal tissues, with an outstanding structural heterogeneity mainly related to molecular weight and sulfation pattern. Recently, few microorganisms, eventually engineered, proved able to synthesize the CS biopolymer backbone, composed of D-glucuronic acid and N-acetyl-D-galactosamine linked through alternating β-(1-3)- and β-(1-4)-glycosidic bonds, and secrete the biopolymers generally unsulfated and possibly decorated with other carbohydrates/molecules. Enzyme catalyzed/assisted methods and chemical tailored protocols allowed to obtain a variety of macromolecules not only resembling the natural extractive ones, but even enlarging the access to unnatural structural features. These macromolecules have been investigated for their bioactivity in vitro and in vivo establishing their potentialities in an array of novel applications in the biomedical field. This review aims to present an overview of the advancements in: i) the metabolic engineering strategies and the biotechnological processes towards chondroitin manufacturing; ii) the chemical approaches applied to obtain specific structural features and targeted decoration of the chondroitin backbone; iii) the biochemical and biological properties of the diverse biotechnological-sourced chondroitin polysaccharides reported so far, unraveling novel fields of applications.

Keywords:  Biotechnological chondroitin; Enzymatic synthesis; Fucosylated chondroitin sulfate; Inflammatory biomarkers; Osteoarthritis in vitro model; Semi-synthetic chondroitin sulfate; Targeted CS substitution pathways; Tissue engineering; Wound healing

DOI:  https://doi.org/10.1016/j.biotechadv.2023.108185
bioRxiv. 2023 May 23. pii: 2023.05.22.541626. [Epub ahead of print]

Chondroitin sulfate, dermatan sulfate, and hyaluronic acid differentially modify the biophysical properties of collagen-based hydrogels.

Marcos Cortes-Medina, Andrew R Bushman, Peter E Beshay, Jonathan J Adorno, Miles M Menyhert, Riley M Hildebrand, Shashwat S Agarwal, Alex Avendano, Jonathan W Song.

Fibrillar collagens and glycosaminoglycans (GAGs) are structural biomolecules that are natively abundant to the extracellular matrix (ECM). Prior studies have quantified the effects of GAGs on the bulk mechanical properties of the ECM. However, there remains a lack of experimental studies on how GAGs alter other biophysical properties of the ECM, including ones that operate at the length scales of individual cells such as mass transport efficiency and matrix microstructure. Here we characterized and decoupled the effects of the GAG molecules chondroitin sulfate (CS) dermatan sulfate (DS) and hyaluronic acid (HA) on the stiffness (indentation modulus), transport (hydraulic permeability), and matrix microarchitecture (pore size and fiber radius) properties of collagen-based hydrogels. We complement these biophysical measurements of collagen hydrogels with turbidity assays to profile collagen aggregate formation. Here we show that CS, DS, and HA differentially regulate the biophysical properties of hydrogels due to their alterations to the kinetics of collagen self-assembly. In addition to providing information on how GAGs play significant roles in defining key physical properties of the ECM, this work shows new ways in which stiffness measurements, microscopy, microfluidics, and turbidity kinetics can be used complementary to reveal details of collagen self-assembly and structure.

DOI: https://doi.org/10.1101/2023.05.22.541626
Angew Chem Int Ed Engl. 2023 Jun 07. e202304325

Rational Design and Expedient Synthesis of Heparan Sulfate Mimetics from Natural Aminoglycosides for Structure and Activity Relationship Studies.

Joseph Wakpal, Vishaka Pathiranage, Alice R Walker, Hien M Nguyen.

  Heparan sulfate (HS) contains variably repeating disaccharide units organized into high and low-sulfated domains. This rich structural diversity enables HS to interact with many proteins and regulate key signaling pathways. Efforts to understand structure-function relationships and harness the therapeutic potential of HS are hindered by the inability to synthesize an extensive library of well-defined HS structures. We herein report a rational and expedient approach to access a library of 27 oligosaccharides from natural aminoglycosides as HS mimetics in 7 - 12 steps. This strategy significantly reduces the number of steps compared to the traditional synthesis of HS oligosaccharides from monosaccharide building blocks. Combined with computational insights, we identify a new class of four trisaccharide compounds derived from aminoglycoside tobramycin that mimic natural HS and have a strong binding to heparanase but a low affinity for off-target platelet factor-4 protein. This work contributes to a better understanding of HS-protein interactions and provides a useful tool for the development of anti-heparanase therapeutics.

Keywords:  Heparan sulfate; aminoglycosides; and platelet factor 4; heparanase; tobramycin

DOI:  https://doi.org/10.1002/anie.202304325
J Agric Food Chem. 2023 Jun 09.

Chondroitin Sulfate Alleviated Obesity by Modulating Gut Microbiota and Liver Metabolome in High-Fat-Diet-Induced Obese Mice.

Ruichang Gao, Zexiu Qi, Jie Lin, Ge Wang, Ge Chen, Li Yuan, Quancai Sun.

  Chondroitin sulfate (CS) is a special bioactive substance with lipid metabolism regulation functions; its molecular mechanisms, however, need further study. This study aimed to study the role of gut microbiota and liver metabolome in the anti-obesity effects of CS. The results demonstrated that CS significantly reduced body weight gain and alleviated insulin resistance and dyslipidemia induced by high-fat diet treatment. Moreover, CS interestingly increased the content of Firmicutes in intestinal microbiota. Further studies showed that there were 11 different metabolites involved in metabolic pathways, including the unsaturated fatty acid biosynthesis pathway, primary bile acid biosynthesis, and taurine and hypotaurine metabolism. In addition, Spearman's correlation analysis indicated that the anti-obesity effect of CS is closely related to liver metabolic regulation. Overall, these results provide a possible molecular mechanism by which CS reduces body weight and lipid accumulation.

Keywords:  chondroitin sulfate; gut microbiota; metabolic disorders; obesity; sturgeon head cartilage

DOI:  https://doi.org/10.1021/acs.jafc.3c02642
J Drugs Dermatol. 2023 Jun 01. 22(6): 546-553

Reduction of Erythema in Moderate-Severe Rosacea by a Low Molecular Weight Heparan Sulfate Analog (HSA).

Rosalyn George, Richard L Gallo, Joel L Cohen, Madeline Brown, Chidubem A V Okeke, Angel S Byrd.

Rosacea changes are a result of an immune mediated response and the angiogenic properties of the LL-37 peptide. This peptide induces an inflammatory signal that activates the NLRP3-mediated inflammasome, triggering rosacea pathogenesis. Research findings show that LL-37 peptide is inhibited by binding to a cell surface glycosaminoglycan, heparan sulfate. Heparan Sulfate Analog (HSA) is a proprietary low molecular weight analog of heparan sulfate that has been formulated into a Dermal Repair Cream (DRC), specifically to aid in such immune mediated responses. Herein, in vitro studies using human epidermal keratinocytes showed an increase in HSA decreased LL-37 toxicity and IL-8 cytokine release. A single-center, randomized double-blind trial included 16 subjects (Fitzpatrick skin types I-IV) with a clinical diagnosis of type 1 rosacea and moderate to severe facial erythema, who were undergoing Pulsed Dye Laser (PDL) treatment. The clinical improvements of their facial erythema were assessed at baseline, 2 weeks, 4 weeks, and 8 weeks. Results revealed that low molecular weight HSA significantly improves the clinical signs of rosacea during the 8 weeks of use likely resulting from inhibition of LL-37 induced IL-8 cytokine release. These findings support the use of DRC in rosacea topical treatment regimens as it demonstrates visible skin benefits and improves tolerability of PDL therapy in a shorter duration of time as compared with PDL alone.George R, Gallo RL, Cohen JL, et al. Reduction of erythema in moderate-severe rosacea by a low molecular weight Heparan Sulfate Analog (HSA). J Drugs Dermatol. 2023;22(6):546-553. doi:10.36849/JDD.7494.

DOI: https://doi.org/10.36849/JDD.7494
Front Mol Biosci. 2023 ;10 1194293

Targeting heparan sulfate-protein interactions with oligosaccharides and monoclonal antibodies.

Miaomiao Li, Lars C Pedersen, Ding Xu.

  Heparan sulfate-binding proteins (HSBPs) are structurally diverse extracellular and membrane attached proteins that interact with HS under normal physiological conditions. Interactions with HS offer an additional level of control over the localization and function of HSBPs, which enables them to behave in a more refined manner. Because all cell signaling events start at the cell membrane, and cell-cell communication relies on translocation of soluble factors across the extracellular matrix, HS occupies an apical position in cellular signal transduction by interacting with hundreds of growth factors, cytokines, chemokines, enzymes, enzyme inhibitors, receptors and adhesion molecules. These extracellular and membrane proteins can play important roles in physiological and pathological conditions. For most HS-binding proteins, the interaction with HS represents an essential element in regulating their normal physiological functions. Such dependence on HS suggests that manipulating HS-protein interactions could be explored as a therapeutic strategy to selectively antagonize/activate HS-binding proteins. In this review, we will discuss current understanding of the diverse nature of HS-HSBP interactions, and the latest advancements in targeting the HS-binding site of HSBPs using structurally-defined HS oligosaccharides and monoclonal antibodies.

Keywords:  crystallization; drug target; heparan sulfate binding proteins; monoclonal antibody; oligosaccharide; specificity

DOI:  https://doi.org/10.3389/fmolb.2023.1194293
bioRxiv. 2023 May 17. pii: 2023.05.16.541066. [Epub ahead of print]

Infection-induced epilepsy is caused by increased expression of chondroitin sulfate proteoglycans in hippocampus and amygdala.

Dipan C Patel, Nathaniel Swift, Bhanu P Tewari, Jack L Browning, Courtney Prim, Lata Chaunsali, Ian Kimbrough, Michelle L Olsen, Harald Sontheimer.

Alterations in the extracellular matrix (ECM) are common in epilepsy, yet whether they are cause or consequence of disease is unknow. Using Theiler's virus infection model of acquired epilepsy we find de novo expression of chondroitin sulfate proteoglycans (CSPGs), a major ECM component, in dentate gyrus (DG) and amygdala exclusively in mice with seizures. Preventing synthesis of CSPGs specifically in DG and amygdala by deletion of major CSPG aggrecan reduced seizure burden. Patch-clamp recordings from dentate granule cells (DGCs) revealed enhanced intrinsic and synaptic excitability in seizing mice that was normalized by aggrecan deletion. In situ experiments suggest that DGCs hyperexcitability results from negatively charged CSPGs increasing stationary cations (K + , Ca 2+ ) on the membrane thereby depolarizing neurons, increasing their intrinsic and synaptic excitability. We show similar changes in CSPGs in pilocarpine-induced epilepsy suggesting enhanced CSPGs in the DG and amygdala may be a common ictogenic factor and novel therapeutic potential.

DOI: https://doi.org/10.1101/2023.05.16.541066
Nat Microbiol. 2023 Jun 05.

Assimilatory sulfate reduction in the marine methanogen Methanothermococcus thermolithotrophicus.

Marion Jespersen, Tristan Wagner.

Methanothermococcus thermolithotrophicus is the only known methanogen that grows on sulfate as its sole sulfur source, uniquely uniting methanogenesis and sulfate reduction. Here we use physiological, biochemical and structural analyses to provide a snapshot of the complete sulfate reduction pathway of this methanogenic archaeon. We find that later steps in this pathway are catalysed by atypical enzymes. PAPS (3'-phosphoadenosine 5'-phosphosulfate) released by APS kinase is converted into sulfite and 3'-phosphoadenosine 5'-phosphate (PAP) by a PAPS reductase that is similar to the APS reductases of dissimilatory sulfate reduction. A non-canonical PAP phosphatase then hydrolyses PAP. Finally, the F420-dependent sulfite reductase converts sulfite to sulfide for cellular assimilation. While metagenomic and metatranscriptomic studies suggest that the sulfate reduction pathway is present in several methanogens, the sulfate assimilation pathway in M. thermolithotrophicus is distinct. We propose that this pathway was 'mix-and-matched' through the acquisition of assimilatory and dissimilatory enzymes from other microorganisms and then repurposed to fill a unique metabolic role.

DOI: https://doi.org/10.1038/s41564-023-01398-8
J Oral Rehabil. 2023 Jun 05.

Sex Hormones associated with Temporomandibular Pain on Palpation in Male Adolescents - Results of the Epidemiologic LIFE Child Study.

Christian Hirsch, Oliver Schierz, Antje Körner, Wieland Kiess, Ronald Biemann, Annett Schrock, Jens Christoph Türp.

  OBJECTIVE: This study aimed to investigate whether the sex steroid precursor hormone dehydroepiandrosterone sulfate (DHEA-S), sex hormone-binding globulin (SHBG), and testosterone (TT) are associated with temporomandibular (TM) pain on palpation in male adolescents.METHODS: Out of the LIFE Child study dataset containing 1,022 children and adolescents aged 10 to 18 years (496 males, 48.5%), we used a subsample of 273 male adolescents (mean age: 13.8 ± 2.3 years) in advanced pubertal development (PD) to analyze the association between hormones and TM pain. The Tanner scale was applied to describe the stage of PD. Pain on palpation of the temporalis and masseter muscles and the TM joints (palpation pain) was assessed using the Diagnostic Criteria for Temporomandibular Disorders (DC/TMD). Serum levels of sex hormones (DHEA-S, SHBG, TT) were determined using standardized laboratory analyses. Free TT was estimated from the ratio between TT and SHBG (free androgen index = FAI). We calculated the risk of perceived positive palpation pain for male participants as a function of hormone levels (DHEA-S, FAI) taking into account age and body mass index (BMI).
RESULTS: Among more developed (Tanner stage 4-5) male adolescents, 22.7% (n=62) reported palpation pain in the TM region. In these participants, FAI levels were approximately half that of individuals without such pain (p<0.01). DHEA-S levels were about 30% lower in the pain group (p<0.01). In multivariable regression analyses, the odds ratio (OR) for pain on palpation decreased to 0.75 (95% confidence interval [CI]: 0.57-0.98) per 10 units of FAI level compared to those without pain, after controlling for the effects of age and adjusted BMI. We observed the same effect for this subgroup per unit of DHEA-S serum level (OR=0.71; 95% CI: 0.53-0.94).
CONCLUSION: At subclinical lower levels of serum free TT and DHEA-S, male adolescents are more likely to report pain on standardized palpation of the masticatory muscles and/or TM joints. This finding supports the hypothesis that sex hormones may influence pain reporting.

Keywords:  male adolescents; palpation pain; sex hormones; temporomandibular disorders

DOI:  https://doi.org/10.1111/joor.13530
Methods Mol Biol. 2023 ;2676 233-243

Site-Specific Incorporation of Sulfotyrosine into Proteins in Mammalian Cells.

Xinyuan He, Yan Chen, Jiantao Guo, Wei Niu.

  Protein tyrosine O-sulfation (PTS) plays a crucial role in numerous extracellular protein-protein interactions. It is involved in diverse physiological processes and the development of human diseases, including AIDS and cancer. To facilitate the study of PTS in live mammalian cells, an approach for the site-specific synthesis of tyrosine-sulfated proteins (sulfoproteins) was developed. This approach takes advantage of an evolved Escherichia coli tyrosyl-tRNA synthetase to genetically encode sulfotyrosine (sTyr) into any proteins of interest (POI) in response to a UAG stop codon. Here, we give a step-by-step account of the incorporation of sTyr in HEK293T cells using the enhanced green fluorescent protein as an example. This method can be widely applied to incorporating sTyr into any POI to investigate the biological functions of PTS in mammalian cells.

Keywords:  Amber suppression; Genetic code expansion; Green fluorescent protein; Mammalian cells; Protein tyrosine O-sulfation (PTS); Sulfotyrosine incorporation

DOI:  https://doi.org/10.1007/978-1-0716-3251-2_17
Front Endocrinol (Lausanne). 2023 ;14 1131438

Association of size for gestational age and dehydroepiandrosterone sulfate with cardiometabolic risk in central precocious puberty girls.

Guijiao Zhang, Huan Yu, Shengxu Yu, Xiaoping Luo, Yan Liang, Ling Hou, Wei Wu.

  Objective: The aim of this study was to assess whether size for gestational age and dehydroepiandrosterone sulfate (DHEAS) are associated with cardiometabolic risk in central precocious puberty (CPP) girls.Methods: The retrospective study included 443 patients with newly diagnosed CPP. Subjects were categorized by birth weight for gestational age (appropriate [AGA], small [SGA], and large [LGA] for gestational age) and serum DHEAS concentration (high [≥75th percentile] and normal [<75th percentile] DHEAS). Cardiometabolic parameters were examined. Composite cardiometabolic risk (CMR) score was calculated based on BMI, blood pressure, glucose, insulin, triglyceride, and HDL cholesterol. Non-obesity CMR score was computed, omitting the value from BMI. Logistic regression models, general linear models, and partial correlation analyses were used to evaluate associations. Propensity score matching was performed for sensitivity analyses.
Results: Overall, 309 patients (69.8%) were born AGA, 80 (18.1%) were born SGA, and 54 (12.2%) were born LGA. Compared with AGA counterparts, CPP girls born SGA were more prone to have elevated HbA1c (adjusted OR = 4.54; 95% CI, 1.43-14.42) and low HDL cholesterol (adjusted OR = 2.33; 95% CI, 1.18-4.61). In contrast, being born LGA was not associated with increased risk for any glucose or lipid derangements. Despite the fact that elevated CMR score was more common among individuals born LGA than AGA (adjusted OR = 1.84; 95% CI, 1.07-4.35), no significant difference was found on non-obesity CMR score (adjusted OR = 0.75; 95% CI, 0.30-1.88). When controlling for age, birth weight SDS, and current BMI-SDS, individuals with high DHEAS exhibited higher HDL cholesterol and apolipoprotein A-1 concentrations and lower triglyceride level and non-obesity CMR score. Furthermore, DHEAS correlated positively with HDL cholesterol and apolipoprotein A-1 and negatively with triglyceride, prominently in girls born SGA, after adjustments for the three abovementioned confounders. Sensitivity analyses corroborated the findings.
Conclusion: Among CPP girls, those born SGA were more likely to possess cardiometabolic risk factors compared to their AGA peers. The difference we observed in cardiometabolic risk between individuals born LGA and AGA was driven by BMI. High DHEAS was associated with favorable lipid profile in CPP girls, even in subjects born SGA.

Keywords:  birth weight for gestational age; cardiometabolic risk; central precocious puberty; children; dehydroepiandrosterone sulfate

DOI:  https://doi.org/10.3389/fendo.2023.1131438
Glycobiology. 2023 Jun 08. pii: cwad044. [Epub ahead of print]

Direct anti-tumoral effects of sulfated fucans isolated from echinoderms: a possible role of neuropilin-1/β1 integrin endocytosis and focal adhesion kinase (FAK) degradation.

Antonio Gilclêr F Lima, Viviane W Mignone, Francisco Vardiero, Eliene O Kozlowski, Laila R Fernandes, Juliana M Motta, Mauro S G Pavão, Camila C Figueiredo, Paulo A S Mourão, Verônica Morandi.

  Hypercoagulability, a major complication of metastatic cancers, has usually been treated with heparins from natural sources, or with their synthetic derivatives, which are under intense investigation in clinical oncology. However, the use of heparin has been challenging for patients with risk of severe bleeding. While the systemic administration of heparins, in pre-clinical models, has shown primarily attenuating effects on metastasis, their direct effect on established solid tumors has generated contradictory outcomes. We investigated the direct antitumoral properties of two sulfated fucans isolated from marine echinoderms, FucSulf1 and FucSulf2, which exhibit anticoagulant activity with mild hemorrhagic potential. Unlike heparin, sulfated fucans significantly inhibited tumor cell proliferation (by ~30-50 percent), and inhibited tumor migration and invasion in vitro. We found that FucSulf1 and FucSulf2 interacted with fibronectin (FN) as efficiently as heparin, leading to loss of prostate cancer and melanoma cell spreading. The sulfated fucans increased the endocytosis of β1 integrin and neuropilin-1 (NRP-1) chains, two cell receptors implicated in FN-dependent adhesion. The treatment of cancer cells with both sulfated fucans, but not with heparin, also triggered intracellular focal adhesion kinase (FAK) degradation, with a consequent overall decrease in activated FAK levels. Finally, only sulfated fucans inhibited the growth of B16-F10 melanoma cells implanted in the dermis of syngeneic C57/BL6 mice. FucSulf1 and FucSulf2 arise from this study as candidates for the design of possible alternatives to long-term treatments of cancer patients with heparins, with the advantage of also controlling local growth and invasion of malignant cells.

Keywords:  Cancer; FAK; fibronectin; integrin endocytosis; sulfated polysaccharides

DOI:  https://doi.org/10.1093/glycob/cwad044
Nature. 2023 Jun 07.

Structural basis for FGF hormone signalling.

Lingfeng Chen, Lili Fu, Jingchuan Sun, Zhiqiang Huang, Mingzhen Fang, Allen Zinkle, Xin Liu, Junliang Lu, Zixiang Pan, Yang Wang, Guang Liang, Xiaokun Li, Gaozhi Chen, Moosa Mohammadi.

α/βKlotho coreceptors simultaneously engage fibroblast growth factor (FGF) hormones (FGF19, FGF21 and FGF23)1,2 and their cognate cell-surface FGF receptors (FGFR1-4) thereby stabilizing the endocrine FGF-FGFR complex3-6. However, these hormones still require heparan sulfate (HS) proteoglycan as an additional coreceptor to induce FGFR dimerization/activation and hence elicit their essential metabolic activities6. To reveal the molecular mechanism underpinning the coreceptor role of HS, we solved cryo-electron microscopy structures of three distinct 1:2:1:1 FGF23-FGFR-αKlotho-HS quaternary complexes featuring the 'c' splice isoforms of FGFR1 (FGFR1c), FGFR3 (FGFR3c) or FGFR4 as the receptor component. These structures, supported by cell-based receptor complementation and heterodimerization experiments, reveal that a single HS chain enables FGF23 and its primary FGFR within a 1:1:1 FGF23-FGFR-αKlotho ternary complex to jointly recruit a lone secondary FGFR molecule leading to asymmetric receptor dimerization and activation. However, αKlotho does not directly participate in recruiting the secondary receptor/dimerization. We also show that the asymmetric mode of receptor dimerization is applicable to paracrine FGFs that signal solely in an HS-dependent fashion. Our structural and biochemical data overturn the current symmetric FGFR dimerization paradigm and provide blueprints for rational discovery of modulators of FGF signalling2 as therapeutics for human metabolic diseases and cancer.

DOI: https://doi.org/10.1038/s41586-023-06155-9
Front Endocrinol (Lausanne). 2023 ;14 1156680

Circulating adrenal 11-oxygenated androgens are associated with clinical outcome in endometrial cancer.

Cylia Dahmani, Patrick Caron, David Simonyan, Véronique Turcotte, Jean Grégoire, Marie Plante, Chantal Guillemette.

  Context: Recent evidence support that androgens play an important role in the etiology of endometrial cancer (EC). Adrenal-derived 11-oxygenated androgens are highly potent agonists of the androgen receptor (AR), comparable to testosterone (T) and dihydrotestosterone (DHT) that have not been studied in the context of EC.Methodology: We studied a cohort of 272 newly diagnosed postmenopausal EC cases undergoing surgical treatment. Circulating concentrations of seven 11-oxygenated androgens including precursors, potent androgens and their metabolites were established in serum samples collected before and 1 month after surgery using a validated liquid chromatography tandem mass spectrometry method (LC-MS/MS). Free (unconjugated) and total (free + sulfate and glucuronide conjugates following enzymatic hydrolysis) were analyzed in relation to clinicopathological features, recurrence and disease-free survival (DFS).
Results: Levels of 11-oxygenated androgens were weakly correlated to those of canonical androgens such as testosterone (T) and dihydrotestosterone (DHT), with no evidence of their association with clinicopathological features. Levels of 11-oxygenated androgens declined after surgery but remained higher in overweight and obese compared to normal weight cases. Higher levels of preoperative free 11-ketoandrosterone (11KAST) were associated with an increased risk of recurrence (Hazard ratio (HR) of 2.99 (95%CI=1.09-8.18); P=0.03). Postoperative free 11β-hydroxyandrosterone (11OHAST) levels were adversely associated with recurrence and DFS (HR = 3.23 (1.11-9.40); P=0.03 and 3.27 (1.34-8.00); P=0.009, respectively).
Conclusion: 11-oxygenated androgen metabolites emerge as potential prognostic markers of EC.

Keywords:  11-oxygenated androgens; endometrial cancer (EC); mass spectrometry - LC-MS/MS; recurrence; survival

DOI:  https://doi.org/10.3389/fendo.2023.1156680
J Extracell Vesicles. 2023 Jun;12(6): e12327

Large-scale heparin-based bind-and-elute chromatography identifies two biologically distinct populations of extracellular vesicles.

Yijun Zhou, Runjie Yuan, Allaura S Cone, Kyle W Shifflett, Gabriel F Arias, Alice Peng, Meredith G Chambers, Ryan P McNamara, Smaranda Willcox, Justin T Landis, Yue Pan, Jack Griffith, Dirk P Dittmer.

  Purifying extracellular vesicles (EVs) has been challenging because EVs are heterogeneous in cargo yet share similar sizes and densities. Most surface marker-based affinity separation methods are limited to research or diagnostic scales. We report that heparin chromatography can separate purified EVs into two distinct subpopulations as ascertained by MS/MS: a non-heparin-binding (NHB) fraction that contains classical EV markers such as tetraspanins and a heparin-binding (HB) fraction enriched in fibronectins and histones. Both fractions were similarly fusogenic but induced different transcriptional responses in endothelial cells. While EVs that were purified by conventional, non-affinity methods alone induced ERK1/2 phosphorylation and Ki67, the NHB fraction did not. This result suggests heparin chromatography as an additional novel fractionation step that is inherently scalable, does not lead to loss of material, and separates inflammatory and pyrogenic EVs from unreactive EVs, which will improve clinical applications.

Keywords:  exomere; exosome; extracellular particles; extracellular vesicles; fibronectin; heparin

DOI:  https://doi.org/10.1002/jev2.12327
ACS Appl Mater Interfaces. 2023 Jun 05.

Heparin Affinity-Based IL-4 Delivery to Modulate Macrophage Phenotype and Endothelial Cell Activity In Vitro.

Fan Zhang, Jessica M Gluck, Ashley C Brown, David A Zaharoff, Martin W King.

  Macrophages play a pivotal role in wound healing and tissue regeneration, as they are rapidly recruited to the site of injury or implanted foreign material. Depending on their interaction with the material, macrophages can develop different phenotypes, with the M1 pro-inflammatory and M2 pro-regenerative phenotypes being highly involved in tissue regeneration. M2 macrophages mitigate inflammation and promote tissue regeneration and extracellular matrix remodeling. In this study, we engineered a gelatin-heparin-methacrylate (GelMA-HepMA) hydrogel that gradually releases interleukin-4 (IL-4), a cytokine that modulates macrophages to adopt the M2 phenotype. Methacrylation of heparin improved the retention of both heparin and IL-4 within the hydrogel. The GelMA-HepMA hydrogel and IL-4 synergistically downregulated M1 gene expression and upregulated M2 gene expression in macrophages within 48 h of in vitro cell culture. However, the M2-like macrophage phenotype induced by the GelMA-HepMA-IL-4 hydrogel did not necessarily further improve endothelial cell proliferation and migration in vitro.

Keywords:  endothelial cell; gelatin methacryloyl; heparin; hydrogel; immunomodulation; interleukin 4; macrophage

DOI:  https://doi.org/10.1021/acsami.3c00489