bims-supasi 2022-11-27 papers

bims-supasi

Biomed News

on Sulfation pathways and signalling

Issue of 2022–11–27
seventeen papers selected by
Jonathan Wolf Mueller, University of Birmingham

Molecular determinants of the interaction between HSV-1 glycoprotein D and heparan sulfate.
Arthritis and Duchenne Muscular Dystrophy: the role of chondroitin sulfate and its associated proteoglycans in disease pathology and as a diagnostic marker.
The Alterations and Roles of Glycosaminoglycans in Human Diseases.
Chitosan, chondroitin sulfate, and hyaluronic acid based in-situ forming scaffold for efficient cell grafting.
Preparation and Multitarget Anti-AD Activity Study of Chondroitin Sulfate Lithium in AD Mice Induced by Combination of D-Gal/AlCl3.
Structure of the human heparan sulfate polymerase complex EXT1-EXT2.
Simvastatin and Dehydroepiandrosterone Sulfate Effects Against Hypoxic Pulmonary Hypertension Are Not Additive.
Opposing Roles of IGFBP-3 and Heparanase in Regulating A549 Lung Cancer Cell Survival.
Total synthesis of chondroitin sulfate E oligosaccharides and biological study.
Enzymatic Digestion of Cell-surface Heparan Sulfate Alters the Radiation Response in Triple-negative Breast Cancer Cells.
Structure of human placental steroid sulfatase at 2.0 angstrom resolution: catalysis, quaternary association, and a secondary ligand site.
CSP I-plus modified rEndostatin inhibits hepatocellular carcinoma metastasis via down-regulation of VEGFA and integrinβ1.
Chondroitin Sulfate Proteoglycan 4 as a Marker for Aggressive Squamous Cell Carcinoma.
Extracellular Heparan 6-O-Endosulfatases SULF1 and SULF2 in Head and Neck Squamous Cell Carcinoma and Other Malignancies.
The relationship of uremic toxin indoxyl sulfate and intestinal elimination mechanisms in hemodialysis patients.
Fabrication of pH Responsive Hydrogel Blends of Chondroitin Sulphate/Pluronic F-127 for the Controlled Release of Ketorolac: Its Characterization and Acute Oral Toxicity Study.
Synthetic Heparan Sulfate Mimetic Polymer Enhances Corneal Nerve Regeneration and Wound Healing after Experimental Laser Ablation Injury in Mice.

Front Mol Biosci. 2022 ;9 1043713

Molecular determinants of the interaction between HSV-1 glycoprotein D and heparan sulfate.

Lauren A Gandy, Ashley J Canning, Huan Lou, Ke Xia, Peng He, Guowei Su, Tina Cairns, Jian Liu, Fuming Zhang, Robert J Linhardt, Gary Cohen, Chunyu Wang.

  Literature has well-established the importance of 3-O-sulfation of neuronal cell surface glycan heparan sulfate (HS) to its interaction with herpes simplex virus type 1 glycoprotein D (gD). Previous investigations of gD to its viral receptors HVEM and nectin-1 also highlighted the conformational dynamics of gD's N- and C-termini, necessary for viral membrane fusion. However, little is known on the structural interactions of gD with HS. Here, we present our findings on this interface from both the glycan and the protein perspective. We used C-terminal and N-terminal gD variants to probe the role of their respective regions in gD/HS binding. The N-terminal truncation mutants (with Δ1-22) demonstrate equivalent or stronger binding to heparin than their intact glycoproteins, indicating that the first 22 amino acids are disposable for heparin binding. Characterization of the conformational differences between C-terminal truncated mutants by sedimentation velocity analytical ultracentrifugation distinguished between the "open" and "closed" conformations of the glycoprotein D, highlighting the region's modulation of receptor binding. From the glycan perspective, we investigated gD interacting with heparin, heparan sulfate, and other de-sulfated and chemically defined oligosaccharides using surface plasmon resonance and glycan microarray. The results show a strong preference of gD for 6-O-sulfate, with 2-O-sulfation becoming more important in the presence of 6-O-S. Additionally, 3-O-sulfation shifted the chain length preference of gD from longer chain to mid-chain length, reaffirming the sulfation site's importance to the gD/HS interface. Our results shed new light on the molecular details of one of seven known protein-glycan interactions with 3-O-sulfated heparan sulfate.

Keywords:  HSV-1; glycoprotein D; heparan sulfate; heparin; herpes

DOI:  https://doi.org/10.3389/fmolb.2022.1043713
Am J Physiol Cell Physiol. 2022 Nov 21.

Arthritis and Duchenne Muscular Dystrophy: the role of chondroitin sulfate and its associated proteoglycans in disease pathology and as a diagnostic marker.

Marissa L Maciej-Hulme, James Melrose, Brooke L Farrugia.

  Chondroitin sulfate (CS) is a ubiquitous glycosaminoglycan covalently attached to the core proteins of cell surface, extracellular, and intracellular proteoglycans. The multistep and highly regulated biosynthesis of chondroitin sulfate, and its degradation products give rise to a diverse species of molecules with functional regulatory properties in biological systems. This review will elucidate and expand on the most recent advances in understanding the role of chondroitin sulfate and its associate proteoglycans, in arthritis and Duchenne muscular dystrophy (DMD), two different and discrete pathologies. Highlighting not only the biodiverse nature of this family of molecules, but the utilization of CS proteoglycans, CS, and its catabolic fragments as biomarkers and potential therapeutic targets for disease pathologies.

Keywords:  Duchene Muscular Dystrophy; arthritis; chondroitin sulfate

DOI:  https://doi.org/10.1152/ajpcell.00103.2022
Polymers (Basel). 2022 Nov 18. pii: 5014. [Epub ahead of print]14(22):

The Alterations and Roles of Glycosaminoglycans in Human Diseases.

Qingchi Wang, Lianli Chi.

  Glycosaminoglycans (GAGs) are a heterogeneous family of linear polysaccharides which are composed of a repeating disaccharide unit. They are also linked to core proteins to form proteoglycans (PGs). GAGs/PGs are major components of the cell surface and the extracellular matrix (ECM), and they display critical roles in development, normal function, and damage response in the body. Some properties (such as expression quantity, molecular weight, and sulfation pattern) of GAGs may be altered under pathological conditions. Due to the close connection between these properties and the function of GAGs/PGs, the alterations are often associated with enormous changes in the physiological/pathological status of cells and organs. Therefore, these GAGs/PGs may serve as marker molecules of disease. This review aimed to investigate the structural alterations and roles of GAGs/PGs in a range of diseases, such as atherosclerosis, cancer, diabetes, neurodegenerative disease, and virus infection. It is hoped to provide a reference for disease diagnosis, monitoring, prognosis, and drug development.

Keywords:  ECM remodeling; chondroitin sulfate; dermatan sulfate; glycosaminoglycan; heparan sulfate; human disease

DOI:  https://doi.org/10.3390/polym14225014
Int J Biol Macromol. 2022 Nov 21. pii: S0141-8130(22)02716-7. [Epub ahead of print]

Chitosan, chondroitin sulfate, and hyaluronic acid based in-situ forming scaffold for efficient cell grafting.

Shrikant Sitaram Kirwale, Swati Sharma, Aniruddha Roy.

  Current cell grafting techniques are majorly dependent on seeding cells on a pre-formed scaffold. However, cells grow in a 2-dimensional (2D) space in such constructs, not mimicking the tissue's 3-dimensional (3D) architecture. The present study evaluated a unique poly-electrolyte complexation (PEC) based strategy for the 3D engraftment of cells in a porous polymeric scaffold. The scaffold was synthesized using a positively charged polysaccharide chitosan (CH) and negatively charged glycosaminoglycans chondroitin sulfate (CS) and hyaluronic acid (HA). Two different scaffolds were synthesized, one using CH and CS [CH-CS] and another using CH and CS + HA [CH-(CS-HA)]. The physicochemical characterization of both the PECs confirmed electrostatic interactions, leading to a porous and viscoelastic PEC formation. Fibroblast cells were grafted and seeded in both scaffolds to evaluate the effect of different scaffold compositions and the difference between seeded and grafted cells. Imaging studies confirmed that grafting of the fibroblast cells supports cellular proliferation. The qPCR studies demonstrated increased expression of functional markers TGF-β, α-SMA, collagen-I, and fibronectin in the CH-(CS-HA) grafted cells. In summary, it was demonstrated that an in-situ forming PEC of CH, CS, and HA had good physicochemical properties for cell grafting and supported grafted cells with improved function.

Keywords:  Cell-grafting; Chitosan; Chondroitin sulfate; Hyaluronic acid; In-situ scaffold; Wound healing

DOI:  https://doi.org/10.1016/j.ijbiomac.2022.11.157
Oxid Med Cell Longev. 2022 ;2022 9466166

Preparation and Multitarget Anti-AD Activity Study of Chondroitin Sulfate Lithium in AD Mice Induced by Combination of D-Gal/AlCl3.

Debo Gao, Pingli Li, Fei Gao, Yangjun Feng, Xiaolin Li, Delong Li, Yuqin Li, Yuliang Xiao.

Previous studies have demonstrated that both CS and LiCl possess anti-Alzheimer's disease (AD) activities. We prepared chondroitin sulfate-Li (CS-Li) and investigated its effect on AD and explored the possible mechanisms both in vitro and in vivo. We found that CS-Li could inhibit amyloid β (Aβ) aggregation and protect SH-SY5Y cells from Aβ 1-42-induced cytotoxicity in vitro. In D-gal and AlCl3-induced AD mouse model, CS-Li improves the spatial learning and memory abilities of AD mice, reverses the nuclear pyknosis and cell edema, and increases the survival rate of neurons in hippocampus of mice. Moreover, CS-Li significantly increased the levels of GSH-Px, Na+/K+-ATPase, and ChAT and decreased the levels of MDA and AchE in AD mice. Western blot results demonstrated that CS-Li could decrease the hyperphosphorylation of tau (Ser396/Ser404) by regulating the expression of p-GSK-3β (Ser9) and PP2A and inhibit the expression of proinflammatory factors through inhibiting NF-κB nuclear translocation by activating the MAPK signaling pathways. In a word, CS-Li can delay AD development through multitarget processes, including Aβ aggregation inhibition, oxidative stress damage, tau hyperphosphorylation, and inflammatory response, thereby improves learning and memory abilities.

DOI: https://doi.org/10.1155/2022/9466166
Nat Commun. 2022 Nov 19. 13(1): 7110

Structure of the human heparan sulfate polymerase complex EXT1-EXT2.

Francisco Leisico, Juneina Omeiri, Christine Le Narvor, Joël Beaudouin, Michael Hons, Daphna Fenel, Guy Schoehn, Yohann Couté, David Bonnaffé, Rabia Sadir, Hugues Lortat-Jacob, Rebekka Wild.

Heparan sulfates are complex polysaccharides that mediate the interaction with a broad range of protein ligands at the cell surface. A key step in heparan sulfate biosynthesis is catalyzed by the bi-functional glycosyltransferases EXT1 and EXT2, which generate the glycan backbone consisting of repeating N-acetylglucosamine and glucuronic acid units. The molecular mechanism of heparan sulfate chain polymerization remains, however, unknown. Here, we present the cryo-electron microscopy structure of human EXT1-EXT2, which reveals the formation of a tightly packed hetero-dimeric complex harboring four glycosyltransferase domains. A combination of in vitro and in cellulo mutational studies is used to dissect the functional role of the four catalytic sites. While EXT1 can catalyze both glycosyltransferase reactions, our results indicate that EXT2 might only have N-acetylglucosamine transferase activity. Our findings provide mechanistic insight into heparan sulfate chain elongation as a nonprocessive process and lay the foundation for future studies on EXT1-EXT2 function in health and disease.

DOI: https://doi.org/10.1038/s41467-022-34882-6
Physiol Res. 2022 Nov 25.

Simvastatin and Dehydroepiandrosterone Sulfate Effects Against Hypoxic Pulmonary Hypertension Are Not Additive.

K Krása, O Vajnerova, J D Uris Ova, M Minaříkova, D Miková, M Srbová, K Chalupský, B Kaftanová, V Hampl.

Pulmonary hypertension is a group of disorders characterized by elevated mean pulmonary artery pressure (mPAP) and pulmonary vascular resistance. To test our hypothesis that combining two drugs useful in experimental pulmonary hypertension, statins and dehydroepiandrosterone sulfate (DHEA S), is more effective than either agent alone, we induced pulmonary hypertension in adult male rats by exposing them to hypoxia (10%O2) for 3 weeks. We treated them with simvastatin (60 mg/l) and DHEA S (100 mg/l) in drinking water, either alone or in combination. Both simvastatin and DHEA S reduced mPAP (froma mean±s.d. of 34.4±4.4 to 27.6±5.9 and 26.7±4.8 mmHg, respectively), yet their combination was not more effective (26.7±7.9 mmHg). Differences in the degree of oxidative stress (indicated by malondialdehydeplasma concentration),the rate of superoxide production (electron paramagnetic resonance), or blood nitric oxide levels (chemiluminescence) did not explain the lack of additivity of the effect of DHEA S and simvastatin on pulmonary hypertension. We propose that the main mechanism of both drugs on pulmonary hypertension could be their inhibitory effect on 3-hydroxy-3-methyl-glutaryl-coenzyme A reductase, which could explain their lack of additivity.
Cells. 2022 Nov 08. pii: 3533. [Epub ahead of print]11(22):

Opposing Roles of IGFBP-3 and Heparanase in Regulating A549 Lung Cancer Cell Survival.

Hind Al Khashali, Jadziah Wareham, Ravel Ray, Ben Haddad, Kai-Ling Coleman, Robert Ranzenberger, Patrick McCombs, Jeffrey Guthrie, Deborah Heyl, Hedeel Guy Evans.

  In this study, we examined the roles of heparanase and IGFBP-3 in regulating A549 and H1299 non-small-cell lung cancer (NSCLC) survival. We found that H1299 cells, known to be p53-null with no expression of IGFBP-3, had higher heparanase levels and activity and higher levels of heparan sulfate (HS) in the media compared to the media of A549 cells. Inhibiting heparanase activity or its expression using siRNA had no effect on the levels of IGFBP-3 in the media of A549 cells, reduced the levels of soluble HS fragments, and led to decreased interactions between IGFBP-3 and HS in the media. HS competed with HA for binding to IGFBP-3 or IGFBP-3 peptide (215-KKGFYKKKQCRPSKGRKR-232) but not the mutant peptide (K228AR230A). HS abolished the cytotoxic effects of IGFBP-3 but not upon blocking HA-CD44 signaling with the anti-CD44 antibody (5F12). Blocking HA-CD44 signaling decreased the levels of heparanase in the media of both A549 and H1299 cell lines and increased p53 activity and the levels of IGFBP-3 in A549 cell media. Knockdown of p53 led to increased heparanase levels and reduced IGFBP-3 levels in A549 cell media while knockdown of IGFBP-3 in A549 cells blocked p53 activity and increased heparanase levels in the media.

Keywords:  CD44; IGFBP-3; extracellular; heparan sulfate; heparanase; hyaluronan; lung cancer; p53; signaling

DOI:  https://doi.org/10.3390/cells11223533
J Asian Nat Prod Res. 2022 Nov 21. 1-7

Total synthesis of chondroitin sulfate E oligosaccharides and biological study.

Shuang Yang, Guang-Yan Zhang, Jin-Yue Zhang, Tian-Qi Li, Zhe-Hui Zhao, Ying-Hong Wang, Ping-Sheng Lei.

  A total synthesis approach of CS-E oligosaccharides was established and a series of derivatives were synthesized. These oligosaccharides were evaluated for a glycosaminoglycan (GAG)-binding protein interaction against cytokines, midkine, and pleiotrophin, by surface-plasmon resonance (SPR) assay. The binding epitopes of oligosaccharides to midkine were mapped using a saturation transfer difference (STD) NMR technique. The groups on the reducing end contributed to binding affinity, and should not be ignored in biological assays. These findings contribute to the structure and activity relationship research and a foundation of understanding that will underpin potential future optimization of this class of oligosaccharides as pharmaceutical agents.

Keywords:  Chondroitin sulfate; midkine; oligosaccharides; pleiotrophin

DOI:  https://doi.org/10.1080/10286020.2022.2146583
Arch Med Res. 2022 Nov 18. pii: S0188-4409(22)00146-1. [Epub ahead of print]

Enzymatic Digestion of Cell-surface Heparan Sulfate Alters the Radiation Response in Triple-negative Breast Cancer Cells.

Stefan Lennard Krautschneider, Fabian M Troschel, Eduardo Vadillo, Hans Theodor Eich, Martin Götte, Nancy Adriana Espinoza-Sánchez, Burkhard Greve.

   BACKGROUND AND AIM: Radiation resistance represents a major challenge in the treatment of breast cancer. As heparan sulfate (HS) chains are known to contribute to tumorigenesis, we aimed to investigate the interplay between HS degradation and radiation response in triple-negative breast cancer (TNBC) cells.
METHODS: HS chains were degraded in vitro as TNBC cells MDA-MB-231 and HCC1806 were treated with heparinase I and III. Subsequently, radioresistance was determined via colony formation assay after doses of 2, 4 and 6 Gy. Cell cycle profile, stem cell characteristics, expression of HS, activation of beta integrins, and apoptosis were determined by flow cytometry. Additionally, cell motility was analyzed via wound-healing assays, and expression and activation of FAK, CDK-6, Src, and Erk1/2 were quantified by western blot pre- and post-irradiation. Finally, the expression of cytokines was analyzed using a cytokine array.
RESULTS: Radiation promoted cell cycle changes, while heparinase treatment induced apoptosis in both cell lines. Colony formation assays showed significantly increased radio-resistance for both cell lines after degradation of HS. Cell migration was similarly upregulated after degradation of HS compared to controls. This effect was even more prominent after irradiation. Interestingly, FAK, a marker of radioresistance, was significantly activated in the heparinase-treated group. Additionally, we found Src to be dysregulated in MDA-MB-231 cells. Finally, we observed differential secretion of GRO, CXCL1, IGFBP1, IL8, Angiogenin, and Osteoprotegerin after HS degradation and radiotherapy.
CONCLUSION: Our results suggest an influence of HS chains on the development of radioresistance in TNBC.

Keywords:  Cell-surface heparan sulfate; Heparan sulfate chains; Heparanase; Radioresistance; Triple-negative Breast cancer

DOI:  https://doi.org/10.1016/j.arcmed.2022.11.004
J Steroid Biochem Mol Biol. 2022 Nov 22. pii: S0960-0760(22)00179-0. [Epub ahead of print] 106228

Structure of human placental steroid sulfatase at 2.0 angstrom resolution: catalysis, quaternary association, and a secondary ligand site.

Debashis Ghosh.

  Human placental estrone (E1)/dehydroepiandrosterone (DHEA) sulfatase (human placental steroid sulfatase; hSTS) is an integral membrane protein of the endoplasmic reticulum. This Ca2+-dependent enzyme catalyzes the hydrolysis of sulfate esters of E1 and DHEA to yield the respective unconjugated steroids, which then act as precursors for the biosynthesis of 17β-estradiol (E2) and dihydrotestosterone (DHT), respectively, the most potent forms of estrogens and androgens. hSTS is a key enzyme for the local production of E2 and DHT in the breast and the prostate. The enzyme is known to be responsible for maintaining high levels of estrogens in the breast tumor cells. The crystal structure of hSTS purified from human placenta has previously been reported at 2.6Å resolution. Here we present the structure of hSTS determined at the superior 2.0Å resolution bringing new clarity to the atomic architecture of the active site. The molecular basis of catalysis and steroid-protein interaction are revisited in light of the new data. We also reexamine the enzyme's quaternary association and its implication on the membrane integration. A secondary ligand binding pocket at the intermolecular interface and adjacent to the active site access channel, buried into the gill of the mushroom-shaped molecule, has been identified. Its role as well as that of a phosphate ion bound to an exposed histidine side chain are examined from the structure-function perspective. Higher resolution data also aids in the tracing of an important loop missing in the previous structure.

Keywords:  breast cancer; conjugated steroid; dehydroepiandrosterone; dihydrotestosterone; endometriosis; estradiol; estrone; prostate cancer; reproductive steroid; steroid sulfate

DOI:  https://doi.org/10.1016/j.jsbmb.2022.106228
BMC Cancer. 2022 Nov 22. 22(1): 1200

CSP I-plus modified rEndostatin inhibits hepatocellular carcinoma metastasis via down-regulation of VEGFA and integrinβ1.

Xueqin Chen, Yan Wang, Hancong Liu, Jingjing Zhang, Jie Wang, Xiaobao Jin, Yan Ma.

   BACKGROUND: In our previous study, N end of the Circumsporozoite protein (CSP I-plus) modified recombinant human Endostatin (rEndostatin, endostar) (rES-CSP) was constructed, which had antiangiogenic capability and bound to hepatocellular carcinoma in vivo and in vitro. In this study, the inhibition of rES-CSP on hepatocellular carcinoma metastasis was verified in vivo and in vitro, and its possible mechanism was explored.
METHODS: Firstly, the impact of rES-CSP on the migration, adhesion of hepatoma cell HCCLM3 was identified by wound healing, transwell, and on metastasis of orthotopic xenograft model was identified in nude mouse. Then the expression of metastasis-associated molecules (MMP2, E-cadherin, integrinβ1) and angiogenesis-related factors (VEGFA) in vitro and in vivo were detected by real-time PCR, western blotting, immunohistochemistry.
RESULTS: Finally, we found that rES-CSP could inhibit the migration and invasion of HCCLM3, and decrease tumor metastasis and growth in nude mouse orthotopic xenograft models. The tumor inhibiting rates of rES-CSP and Endostar were 42.46 ± 5.39% and 11.1 ± 1.88%. The lung metastasis rates of the control, Endostar and rES-CSP were 71, 50, and 42.8%, respectively. Compared with Endostar, rES-CSP significantly down-regulated the expression of VEGFA and integrinβ1. Heparin, a competitive inhibitor of CSP I-plus, which can be bind to the highly-sulfated heparan sulfate proteoglycans (HSPGs) over-expressed in liver and hepatocellular carcinoma, alleviated the down-regulation of VEGFA and integrinβ1.
CONCLUSIONS: These indicate that rES-CSP may play a role in inhibiting tumor growth and metastasis by down-regulating the angiogenic factor VEGF and the metastasis-related molecules or by interfering with HSPGs-mediated tumor metastasis.

Keywords:  Hepatocellular carcinoma; Liver-targeting peptide CSP I-plus; Tumor metastasis; rEndostatin

DOI:  https://doi.org/10.1186/s12885-022-10318-8
Cancers (Basel). 2022 Nov 13. pii: 5564. [Epub ahead of print]14(22):

Chondroitin Sulfate Proteoglycan 4 as a Marker for Aggressive Squamous Cell Carcinoma.

Kathryn Chen, Joel Yong, Roland Zauner, Verena Wally, John Whitelock, Mila Sajinovic, Zlatko Kopecki, Kang Liang, Kieran Francis Scott, Albert Sleiman Mellick.

  Chondroitin sulfate (CS) proteoglycan 4 (CSPG4) is a cell surface proteoglycan that is currently under investigation as a marker of cancer malignancy, and as a potential target of anticancer drug treatment. CSPG4 acts as a driver of tumourigenesis by regulating turnover of the extracellular matrix (ECM) to promote tumour cell invasion, migration as well as inflammation and angiogenesis. While CSPG4 has been widely studied in certain malignancies, such as melanoma, evidence is emerging from global gene expression studies, which suggests a role for CSPG4 in squamous cell carcinoma (SCC). While relatively treatable, lack of widely agreed upon diagnostic markers for SCCs is problematic, especially for clinicians managing certain patients, including those who are aged or infirm, as well as those with underlying conditions such as epidermolysis bullosa (EB), for which a delayed diagnosis is likely lethal. In this review, we have discussed the structure of CSPG4, and quantitatively analysed CSPG4 expression in the tissues and pathologies where it has been identified to determine the usefulness of CSPG4 expression as a diagnostic marker and therapeutic target in management of malignant SCC.

Keywords:  cancer biomarker; chondroitin sulfate proteoglycan; cutaneous squamous cell carcinoma; gene expression; head and neck squamous cell carcinoma

DOI:  https://doi.org/10.3390/cancers14225564
Cancers (Basel). 2022 Nov 11. pii: 5553. [Epub ahead of print]14(22):

Extracellular Heparan 6-O-Endosulfatases SULF1 and SULF2 in Head and Neck Squamous Cell Carcinoma and Other Malignancies.

Yang Yang, Jaeil Ahn, Nathan J Edwards, Julius Benicky, Aaron M Rozeboom, Bruce Davidson, Christina Karamboulas, Kevin C J Nixon, Laurie Ailles, Radoslav Goldman.

  Pan-cancer analysis of TCGA and CPTAC (proteomics) data shows that SULF1 and SULF2 are oncogenic in a number of human malignancies and associated with poor survival outcomes. Our studies document a consistent upregulation of SULF1 and SULF2 in HNSC which is associated with poor survival outcomes. These heparan sulfate editing enzymes were considered largely functional redundant but single-cell RNAseq (scRNAseq) shows that SULF1 is secreted by cancer-associated fibroblasts in contrast to the SULF2 derived from tumor cells. Our RNAScope and patient-derived xenograft (PDX) analysis of the HNSC tissues fully confirm the stromal source of SULF1 and explain the uniform impact of this enzyme on the biology of multiple malignancies. In summary, SULF2 expression increases in multiple malignancies but less consistently than SULF1, which uniformly increases in the tumor tissues and negatively impacts survival in several types of cancer even though its expression in cancer cells is low. This paradigm is common to multiple malignancies and suggests a potential for diagnostic and therapeutic targeting of the heparan sulfatases in cancer diseases.

Keywords:  SULF1; SULF2; cancer associated fibroblast; head and neck squamous cell carcinoma; heparan sulfate 6-O-endosulfatase

DOI:  https://doi.org/10.3390/cancers14225553
Kidney Blood Press Res. 2022 Nov 24.

The relationship of uremic toxin indoxyl sulfate and intestinal elimination mechanisms in hemodialysis patients.

Adéla Tomášová, Alena Tichá, Sylvie Dusilová Sulková, Roman Safránek, Petr Moučka, Marcela Chmelařová, Ivana Baranová, Helena Párová, Zora Nývltová, Karolína Štochlová, Vladimír Palička, Ladislava Pavlíková, Zdeněk Zadák, Radomír Hyšpler.

INTRODUCTION: High indoxyl sulfate (IS) concentration is a serious problem for patients with CKD increasing the risk of cardiovascular diseases and CKD progression. Thus, the methods of decreasing the toxin concentrations are highly desired. The study aimed to discover the role of selected intestine related factors on IS concentration.
METHODS: We evaluated the impact of ABCG2 and ABCC2 polymorphisms influencing activity and protein intake by normalized protein catabolic rate. Additionally, we examined the relation of IS and uric acid (UA), that can share common elimination transporters. A monocentric, prospective, open cohort pilot study was performed on 108 patients undergoing dialysis treatment.
RESULTS: The positive effect of residual diuresis on the reduction of IS levels was confirmed (p = 0.005). Also, an increase in IS depending on the dietary protein intake was confirmed (p = 0.040). No significant correlation between ABC gene polymorphisms was observed either, suggesting the negligible role of ABCG2 and ABCC2 in the elimination of IS in small bowel. The significant difference was observed for UA where ABCG2 421C>A (rs72552713) gene polymorphism was higher (505.3 µmol/L) in comparison with a wild type genotype (360.5 µmol/L). Discussion/ Conclusion: No evidence of bowel elimination pathway via ABCC2 and ABCG2 transporters was found in renal replacement therapy patients.

DOI: https://doi.org/10.1159/000528130
Drug Dev Ind Pharm. 2022 Nov 24. 1-29

Fabrication of pH Responsive Hydrogel Blends of Chondroitin Sulphate/Pluronic F-127 for the Controlled Release of Ketorolac: Its Characterization and Acute Oral Toxicity Study.

Saba Naeem, Kashif Barkat, Maryam Shabbir, Ikrima Khalid, Irfan Anjum, Nadia Shamshad, Yasir Mehmood, Daulat Haleem Khan, Syed Faisal Badshah, Muhammad Ali Syed, Sana Afzal, Zainab Abdullah.

   OBJECTIVE: Ketorolac tromethamine (KT), selected as a model drug, is used in management of moderate to severe acute pain. It has a short half-life (∼5.5 hrs) and requires frequent dose administration when needed for longer period of time. In our current project we designed pH responsive hydrogel blends of chondroitin sulphate/pluronic F-127 for the controlled release of ketorolac.
METHODS: Hydrogel blends were fabricated using free radical polymerization reaction technique utilizing different ratios of chondroitin sulfate (CS) (polymer) and pluronic F-127 (polymer), acrylic acid (monomer), N-N'-methyl-bisacrylamide (MBA) (cross-linker), initiator ammonium persulfate (APS) and tween-80 (surfactant). The fabricated hydrogel blends were studied and evaluated for pH responsiveness, swelling, water absorbency, in-vitro drug release and morphological characteristics such as SEM, XRD, FTIR and TGA/DSC. Acute toxicity study was performed on rabbits.
RESULTS: Maximum swelling and water absorbency was shown by chondroitin sulfate/pluronic F-127 (CS/Pl) blends being significantly greater at 7.4 (basic pH) than in 1.2 (acidic pH). In-vitro dissolution demonstrated pH responsive controlled KT release following zero order at higher pH (7.4) medium up to 36 hours. FTIR studies confirmed the structures of our blends, SEM results showed porous framework, thermal studies revealed higher stability of hydrogels than the individual polymers, XRD confirmed the nature of our blends. Toxicity study revealed the non-toxic nature of the hydrogel blends.
CONCLUSION: The prepared CS/Pl hydrogels demonstrated stimuli-controlled release with delivery of drug for prolonged period of time and thus can minimize dosing frequency, safe drug delivery, increased patient compliance and easiness.

Keywords:  Chondroitin sulfate; Controlled release; Free radical polymerization; Ketorolac tromethamine; pH responsive

DOI:  https://doi.org/10.1080/03639045.2022.2150773
Polymers (Basel). 2022 Nov 15. pii: 4921. [Epub ahead of print]14(22):

Synthetic Heparan Sulfate Mimetic Polymer Enhances Corneal Nerve Regeneration and Wound Healing after Experimental Laser Ablation Injury in Mice.

Ignacio Alcalde, Cristina Sánchez-Fernández, Susana Del Olmo-Aguado, Carla Martín, Céline Olmiere, Enol Artime, Luis M Quirós, Jesús Merayo-Lloves.

  (1) Background: Abnormal corneal wound healing compromises visual acuity and can lead to neuropathic pain. Conventional treatments usually fail to restore the injured corneal tissue. In this study, we evaluated the effectiveness of a synthetic heparan sulfate mimetic polymer (HSmP) in a mouse model of corneal wound healing. (2) Methods: A surgical laser ablation affecting the central cornea and subbasal nerve plexus of mice was used as a model of the wound-healing assay. Topical treatment with HSmP was contrasted to its vehicle and a negative control (BSS). Corneal repair was studied using immunofluorescence to cell proliferation (Ki67), apoptosis (TUNEL assay), myofibroblast transformation (αSMA), assembly of epithelial cells (E-cadherin) and nerve regeneration (β-tubulin III). (3) Results: At the end of the treatment, normal epithelial cytoarchitecture and corneal thickness were achieved in HSmP-treated animals. HSmP treatment reduced myofibroblast occurrence compared to eyes irrigated with vehicle (p < 0.01) or BSS (p < 0.001). The HSmP group showed 50% more intraepithelial nerves than the BSS or vehicle groups. Only HSmP-treated corneas improved the visual quality to near transparent. (4) Conclusions: These results suggest that HSmP facilitates the regeneration of the corneal epithelium and innervation, as well as restoring transparency and reducing myofibroblast scarring after laser experimental injury.

Keywords:  cornea; extracellular matrix; heparan sulfate mimetic polymer; nerve regeneration; transparency; wound healing

DOI:  https://doi.org/10.3390/polym14224921