bims-supasi 2024-09-15 papers

bims-supasi

Biomed News

on Sulfation pathways and signalling

Issue of 2024–09–15
fourteen papers selected by
Jonathan Wolf Mueller, University of Birmingham

Synthesis of a deuterated disaccharide internal standard for LC-MS/MS quantitation of heparan sulfate in biological samples.
Polysaccharide-based chondroitin sulfate macromolecule loaded hydrogel/scaffolds in wound healing- A comprehensive review on possibilities, research gaps, and safety assessment.
Anti-inflammatory and anti-lung cancer activities of low-molecular-weight and high-sulfate-content sulfated polysaccharides extracted from the edible fungus Poria cocos.
Structural Insights into Endostatin-Heparan Sulfate Interactions Using Modeling Approaches.
Syndecan-1 Plays a Role in the Pathogenesis of Sjögren's Disease by Inducing B-Cell Chemotaxis through CXCL13-Heparan Sulfate Interaction.
Anti-enterovirus 71 activity of native fucosylated chondroitin sulfates and their derivatives.
Chondroitin Sulfate Proteoglycan 4 (CSPG4) as a Potential Biomarker for Fetal Growth Restriction: Insights from an Umbilical Cord Blood Analysis.
Genetic Variance in Heparan Sulfation Is Associated With Salt Sensitivity.
Redesigned chondroitinase ABC degrades inhibitory chondroitin sulfate proteoglycans in vitro and in vivo in the stroke-injured rat brain.
Preparation and characterization of kelp polysaccharide and its research on anti-influenza a virus activity.
Developing and Benchmarking Sulfate and Sulfamate Force Field Parameters via Ab Initio Molecular Dynamics Simulations To Accurately Model Glycosaminoglycan Electrostatic Interactions.
The simultaneous inhibition of SLC6A19 and BCRP transporters leads to an increase of indoxyl sulfate (a uremic toxin) in plasma and kidney.
Cancer-Associated Fibroblasts Expressing Sulfatase 1 Facilitate VEGFA-Dependent Microenvironmental Remodeling to Support Colorectal Cancer.
Acrylamide exposure, sex hormones, and pubertal status in Japanese adolescents.

Carbohydr Res. 2024 Sep 07. pii: S0008-6215(24)00249-0. [Epub ahead of print]545 109270

Synthesis of a deuterated disaccharide internal standard for LC-MS/MS quantitation of heparan sulfate in biological samples.

Yuchen Pan, Qi Qi He, Norbert Wimmer, Vito Ferro.

  High levels of heparan sulfate (HS) are a marker for several mucopolysaccharidosis (MPS) disorders which are lysosomal storage diseases caused by genetic defects in HS-degrading enzymes. Quantitation of HS in biological samples is therefore critical for diagnosis and evaluating the efficacy of new therapies. Herein we present the efficient synthesis of a butylated GlcN-GlcA disaccharide and its deuterated derivative for use as an internal standard in a quantitative mass spectrometry-based assay for analysis of HS following butanolysis. The synthesis features the stereoselective 1,2-cis glycosylation of a GlcA acceptor with a 6-O-benzoyl-2-deoxy-2-azido thioglucoside donor.

Keywords:  Butanolysis; Disaccharide synthesis; Heparan sulfate; Mucopolysaccharidosis

DOI:  https://doi.org/10.1016/j.carres.2024.109270
Int J Biol Macromol. 2024 Sep 06. pii: S0141-8130(24)06218-4. [Epub ahead of print]279(Pt 3): 135410

Polysaccharide-based chondroitin sulfate macromolecule loaded hydrogel/scaffolds in wound healing- A comprehensive review on possibilities, research gaps, and safety assessment.

Shubhrajit Mantry, Ashutosh Behera, Shaktiprasad Pradhan, Lalatendu Mohanty, Ragni Kumari, Ankita Singh, Mahesh Kumar Yadav.

  Wound healing is an intricate multifactorial process that may alter the extent of scarring left by the wound. A substantial portion of the global population is impacted by non-healing wounds, imposing significant financial burdens on the healthcare system. The conventional dosage forms fail to improve the condition, especially in the presence of other morbidities. Thus, there is a pressing requirement for a type of wound dressing that can safeguard the wound site and facilitate skin regeneration, ultimately expediting the healing process. In this context, Chondroitin sulfate (CS), a sulfated glycosaminoglycan material, is capable of hydrating tissues and further promoting the healing. Thus, this comprehensive review article delves into the recent advancement of CS-based hydrogel/scaffolds for wound healing management. The article initially summarizes the various physicochemical characteristics and sources of CS, followed by a brief understanding of the importance of hydrogel and CS in tissue regeneration processes. This is the first instance of such a comprehensive summarization of CS-based hydrogel/scaffolds in wound healing, focusing more on the mechanistic wound healing process, furnishing the recent innovations and toxicity profile. This contemporary review provides a profound acquaintance of strategies for contemporary challenges and future direction in CS-based hydrogel/scaffolds for wound healing.

Keywords:  Chondroitin sulfate; Hydrogel; Macromolecules; Polysaccharides; Scaffolds; Wound healing

DOI:  https://doi.org/10.1016/j.ijbiomac.2024.135410
Int J Biol Macromol. 2024 Sep 10. pii: S0141-8130(24)06291-3. [Epub ahead of print]279(Pt 4): 135483

Anti-inflammatory and anti-lung cancer activities of low-molecular-weight and high-sulfate-content sulfated polysaccharides extracted from the edible fungus Poria cocos.

Wei-Lun Qiu, Chi-Hsein Chao, Mei-Kuang Lu.

  Sulfated polysaccharides (SPSs) have excellent physicochemical properties, attracting research interest in the pharmaceutical industry. A previous study extracted SPS (named Suc40) from the edible fungus, Poria cocos and demonstrated that it exhibited anti-inflammatory and anticancer activities. In this study, three fractions of Suc40, Suc40 F1, Suc40 F2, and Suc40 F3, with different molecular weights and sulfate contents were prepared through gel-filtration column chromatography. The molecular weights of F1, F2, and F3 were approximately 616.23, 82.57, and 6.21 kDa, respectively, and their sulfate content were 0.23, 1.65, and 1.90 mmol/g, respectively. The fractions' anti-inflammatory activities were determined by assessing their ability to suppress inflammatory cytokines in lipopolysaccharide (LPS)-stimulated RAW264.7 cells. Suc40 F2 and Suc40 F3 suppressed interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) production by 60 % and 35 %, respectively. Suc40 F2 and Suc40 F3 suppressed protein kinase B (AKT)/p38 and p38 signaling, which resulted in anti-inflammatory effects. The fractions' anti-lung cancer activity was evaluated by assessing their H1975 cell proliferation inhibition. Suc40 F3 at a concentration of 800 μg/ml exhibited maximal cell proliferation inhibition. The low molecular weight and high sulfate content of Suc40 F3 were associated with its enhanced anti-inflammatory and anti-lung cancer activities.

Keywords:  Molecular weight; Poria cocos; Sulfated polysaccharide; Sulfation

DOI:  https://doi.org/10.1016/j.ijbiomac.2024.135483
Molecules. 2024 Aug 26. pii: 4040. [Epub ahead of print]29(17):

Structural Insights into Endostatin-Heparan Sulfate Interactions Using Modeling Approaches.

Urszula Uciechowska-Kaczmarzyk, Martin Frank, Sergey A Samsonov, Martyna Maszota-Zieleniak.

  Glycosaminoglycans (GAGs) play a key role in a variety of biological processes in the extracellular matrix (ECM) via interactions with their protein targets. Due to their high flexibility, periodicity and electrostatics-driven interactions, GAG-containing complexes are very challenging to characterize both experimentally and in silico. In this study, we, for the first time, systematically analyzed the interactions of endostatin, a proteolytic fragment of collagen XVIII known to be anti-angiogenic and anti-tumoral, with heparin (HP) and representative heparan sulfate (HS) oligosaccharides of various lengths, sequences and sulfation patterns. We first used conventional molecular docking and a docking approach based on a repulsive scaling-replica exchange molecular dynamics technique, as well as unbiased molecular dynamic simulations, to obtain dynamically stable GAG binding poses. Then, the corresponding free energies of binding were calculated and the amino acid residues that contribute the most to GAG binding were identified. We also investigated the potential influence of Zn2+ on endostatin-HP complexes using computational approaches. These data provide new atomistic details of the molecular mechanism of HP's binding to endostatin, which will contribute to a better understanding of its interplay with proteoglycans at the cell surface and in the extracellular matrix.

Keywords:  endostatin; glycosaminoglycans; molecular docking; protein–glycosaminoglycan interactions; repulsive scaling–replica exchange molecular dynamics

DOI:  https://doi.org/10.3390/molecules29174040
Int J Mol Sci. 2024 Aug 29. pii: 9375. [Epub ahead of print]25(17):

Syndecan-1 Plays a Role in the Pathogenesis of Sjögren's Disease by Inducing B-Cell Chemotaxis through CXCL13-Heparan Sulfate Interaction.

Nan Young Lee, Hirut Yadeta Ture, Eun Ju Lee, Ji Ae Jang, Gunwoo Kim, Eon Jeong Nam.

  In Sjögren's disease (SjD), the salivary glandular epithelial cells can induce the chemotaxis of B cells by secreting B-cell chemokines such as C-X-C motif chemokine ligand 13 (CXCL13). Syndecan-1 (SDC-1) is a major transmembrane heparan sulfate proteoglycan (HSPG) predominantly expressed on epithelial cells that binds to and regulates heparan sulfate (HS)-binding molecules, including chemokines. We aimed to determine whether SDC-1 plays a role in the pathogenesis of SjD by acting on the binding of HS to B-cell chemokines. To assess changes in glandular inflammation and SDC-1 concentrations in the submandibular gland (SMG) and blood, female NOD/ShiLtJ and sex- and age-matched C57BL/10 mice were used. In the SMG of NOD/ShiLtJ mice, inflammatory responses were identified at 8 weeks of age, but increased SDC-1 concentrations in the SMG and blood were observed at 6 weeks of age, when inflammation had not yet started. As the inflammation of the SMG worsened, the SDC-1 concentrations in the SMG and blood increased. The expression of the CXCL13 and its receptor C-X-C chemokine receptor type 5 (CXCR5) began to increase in the SMG at 6 weeks of age and continued until 12 weeks of age. Immunofluorescence staining in SMG tissue and normal murine mammary gland cells confirmed the co-localization of SDC-1 and CXCL13, and SDC-1 formed a complex with CXCL13 in an immunoprecipitation assay. Furthermore, NOD/ShiLtJ mice were treated with 5 mg/kg HS intraperitoneally thrice per week for 6-10 weeks of age, and the therapeutic effects in the SMG were assessed at the end of 10 weeks of age. NOD/ShiLtJ mice treated with HS showed attenuated salivary gland inflammation with reduced B-cell infiltration, germinal center formation and CXCR5 expression. These findings suggest that SDC-1 plays a pivotal role in the pathogenesis of SjD by binding to CXCL13 through the HS chain.

Keywords:  B cell; CXCL13; Sjögren’s disease; epithelial cell; syndecan-1

DOI:  https://doi.org/10.3390/ijms25179375
Carbohydr Polym. 2024 Dec 15. pii: S0144-8617(24)00883-X. [Epub ahead of print]346 122657

Anti-enterovirus 71 activity of native fucosylated chondroitin sulfates and their derivatives.

Qingfeng Niu, Han Zhou, Xiaoyao Ma, Yuanyuan Jiang, Chanjuan Liu, Wei Wang, Guangli Yu, Guoyun Li.

  Enterovirus 71 (EV71) is recognized as a major causative agent of hand, foot, and mouth disease (HFMD), posing a significant global public health concern due to its widespread impact and resulting in a major public health issue worldwide. Despite its prevalence, current clinical therapy lacks effective antiviral agents. Fucosylated chondroitin sulfates (FCS) derived from sea cucumber exhibits a range of biological activities including potent antiviral effects. This study provides compelling evidence of the potent antiviral efficacy of FCS against EV71. To further elucidate the impact of structural variations on the anti-EV71 activity, native FCSs with diverse sulfation patterns and a varity of FCS derivatives were prepared and analyzed. Notably, this study presents the detailed structural characterization of FCSs from the sea cucumbers Holothuria scabra Jaege and Holothuria fuscopunctata. Analysis of the structure-activity relationships revealed that molecular weight, sulfated fucose branches, and sulfation pattern were all crucial factors contributing to the potent inhibitory effects of FCS against EV71. Interestingly, molecular weight emerged as the most significant structural determinant of the antiviral potency. These findings suggest the promising potential of utilizing FCS as an innovative EV71 entry inhibitor for the treatment of HFMD.

Keywords:  Antiviral effect; Enterovirus 71; Fucosylated chondroitin sulfate; Structure elucidation; Structure-activity relationship

DOI:  https://doi.org/10.1016/j.carbpol.2024.122657
Clin Lab. 2024 Sep 01. 70(9):

Chondroitin Sulfate Proteoglycan 4 (CSPG4) as a Potential Biomarker for Fetal Growth Restriction: Insights from an Umbilical Cord Blood Analysis.

Zeynep Seyhanli, Burak Bayraktar, Gulsan Karabay, Betul T Cakir, Mevlut Bucak, Gizem Aktemur, Can O Ulusoy, Serap Sucu, Seray Ozturk, Gulcan K Hekimoglu, Sevki Celen.

BACKGROUND: This study aimed to evaluate the umbilical cord blood chondroitin sulfate proteoglycan 4 (CSPG4) concentrations in pregnancies complicated with fetal growth restriction (FGR) and aimed to investigate the rela-tionship between the CSPG4 levels in these pregnancies and adverse neonatal outcomes.
METHODS: This prospective case-control study was conducted between August 2023 and January 2024. The study included 80 singleton pregnancies at 35 to 39 weeks of gestation. Among these, 40 were diagnosed with FGR and 40 served as the control group. After the delivery, samples of the cord blood were collected prior to the placental delivery.
RESULTS: The CSPG4 levels were significantly higher in the study group (FGR), 1,153 (1,059 - 1,261) pg/mL, than in the control group, 1,107 (873 - 1,197) pg/mL (p = 0.024). When all patients were evaluated, the CSPG4 levels showed a positive correlation with the systolic/diastolic (S/D) ratio of the umbilical arteries (r = 0.276, p = 0.013). A statistically significant negative correlation was observed between the levels of CSPG4 in the umbilical cord blood and the Apgar scores at the 1st (r = -0.256, p = 0.022) and 5th (r = -0.250, p = 0.026) minutes. The discriminatory power of the umbilical cord CSPG4 level in the determination of composite adverse neonatal outcomes was evaluated by ROC analysis and a cutoff point of > 1,091.25 pg/mL, showing a sensitivity of 93.3%, a specificity of 46.2%, and an AUC of 0.661 (95% CI: 0.547 - 0.763, p = 0.019).
CONCLUSIONS: Elevated levels of CSPG4 have been observed in the umbilical cord blood in pregnancies complicated by FGR; higher levels are associated with adverse neonatal outcomes.

DOI: https://doi.org/10.7754/Clin.Lab.2024.240436
Hypertension. 2024 Sep 09.

Genetic Variance in Heparan Sulfation Is Associated With Salt Sensitivity.

Jetta J Oppelaar, Bart Ferwerda, Mohamed A Romman, Ghazalah N Sahebdin, Aeilko H Zwinderman, Henrike Galenkamp, S Matthijs Boekholdt, Bert-Jan H van den Born, Rik H G Olde Engberink, Liffert Vogt.

   BACKGROUND: High heritability of salt sensitivity suggests an essential role for genetics in the relationship between sodium intake and blood pressure (BP). The role of glycosaminoglycan genes, which are crucial for salinity tolerance, remains to be elucidated.
METHODS: Interactions between 54 126 variants in 130 glycosaminoglycan genes and daily sodium excretion on BP were explored in 20 420 EPIC-Norfolk (European Prospective Investigation Into Cancer in Norfolk) subjects. The UK Biobank (n=414 132) and the multiethnic HELIUS study (Healthy Life in an Urban Setting; n=2239) were used for validation. Afterward, the urinary glycosaminoglycan composition was studied in HELIUS participants (n=57) stratified by genotype and upon dietary sodium loading in a time-controlled crossover intervention study (n=12).
RESULTS: rs2892799 in NDST3 (heparan sulfate N-deacetylase/N-sulfotransferase 3) showed the strongest interaction with sodium on mean arterial pressure (false discovery rate 0.03), with higher mean arterial pressure for the C allele in high sodium conditions. Also, rs9654628 in HS3ST5 (heparan sulfate-glucosamine 3-sulfotransferase 5) showed an interaction with sodium on systolic BP (false discovery rate 0.03). These interactions were multiethnically validated. Stratifying for the rs2892799 genotype showed higher urinary expression of N-sulfated heparan sulfate epitope D0S0 for the T allele. Conversely, upon dietary sodium loading, urinary D0S0 expression was higher in participants with stable BP after sodium loading, and sodium-induced effects on this epitope were opposite in individuals with and without BP response to sodium.
CONCLUSIONS: The C allele of rs2892799 in NDST3 exhibits higher BP in high sodium conditions when compared with low sodium conditions, whereas no differences were detected for the T allele. Concomitantly, both alleles demonstrate distinct expressions of D0S0, which, in turn, correlates with sodium-mediated BP elevation. These findings underscore the potential significance of genetic glycosaminoglycan variation in human BP regulation.

Keywords:  UK Biobank; blood pressure; glycosaminoglycans; salt tolerance; sodium

DOI:  https://doi.org/10.1161/HYPERTENSIONAHA.124.23421
Biomaterials. 2024 Sep 07. pii: S0142-9612(24)00352-1. [Epub ahead of print]314 122818

Redesigned chondroitinase ABC degrades inhibitory chondroitin sulfate proteoglycans in vitro and in vivo in the stroke-injured rat brain.

Nitzan Letko Khait, Sabrina Zuccaro, Dhana Abdo, Hong Cui, Ricky Siu, Eric Ho, Cindi M Morshead, Molly S Shoichet.

  Injuries to the central nervous system, such as stroke and traumatic spinal cord injury, result in an aggregate scar that both limits tissue degeneration and inhibits tissue regeneration. The aggregate scar includes chondroitin sulfate proteoglycans (CSPGs), which impede cell migration and axonal outgrowth. Chondroitinase ABC (ChASE) is a potent yet fragile enzyme that degrades CSPGs, and thus may enable tissue regeneration. ChASE37, with 37-point mutations to the native enzyme, has been shown to be more stable than ChASE, but its efficacy has never been tested. To answer this question, we investigated the efficacy of ChASE37 first in vitro using human cell-based assays and then in vivo in a rodent model of stroke. We demonstrated ChASE37 degradation of CSPGs in vitro and the consequent cell adhesion and axonal sprouting now possible using human induced pluripotent stem cell (hiPSC)-derived neurons. To enable prolonged release of ChASE37 to injured tissue, we expressed it as a fusion protein with a Src homology 3 (SH3) domain and modified an injectable, carboxymethylcellulose (CMC) hydrogel with SH3-binding peptides (CMC-bp) using inverse electron-demand Diels-Alder chemistry. We injected this affinity release CMC-bp/SH3-ChASE37 hydrogel epicortically to endothelin-1 stroke-injured rats and confirmed bioactivity via degradation of CSPGs and axonal sprouting in and around the lesion. With CSPG degradation shown both in vitro by greater cell interaction and in vivo with local delivery from a sustained release formulation, we lay the foundation to test the potential of ChASE37 and its delivery by local affinity release for tissue regeneration after stroke.

Keywords:  Aggregate scar; Chondroitinase ABC; Human induced pluripotent stem cells; Hydrogel; Stroke

DOI:  https://doi.org/10.1016/j.biomaterials.2024.122818
Int J Biol Macromol. 2024 Sep 09. pii: S0141-8130(24)06314-1. [Epub ahead of print] 135506

Preparation and characterization of kelp polysaccharide and its research on anti-influenza a virus activity.

Tianxiang Pi, Lishan Sun, Wei Li, Wei Wang, Minghui Dong, Xinxing Xu, He Xu, Yuanhui Zhao.

  The beneficial effects of kelp polysaccharide (KPS) have recently attracted attention. In this study, KPS was extracted from kelp using the enzyme hydrolysis combined with freeze-drying, namely, KPS-EF. The structural characterization showed that KPS-EF was a highly sulfated macromolecule with the Mw of 764.2 kDa and the sulfate content of 23.49 %. The antiviral activity of KPS-EF in vitro was verified, and the IC50 value of KPS against the PR8 virus was 0.58 mg/mL. Intranasal administration of KPS-EF significantly inhibited death and weight loss in IAV-infected mice and alleviated virus-induced pneumonia symptoms, meanwhile, KPS-EF (10 mg/kg/day) significantly decreased the production levels of chemokines (CXCL1, RANTES) and inflammatory cytokines (IL-6, TNF-α) in lungs (p < 0.05). KPS-EF could downregulate the activity of viral neuraminidase (NA) primarily in the late stage of viral adsorption with an IC50 value of 0.29 mg/mL. This study provides a theoretical basis for the using KPS as a supplement to NA inhibitors or anti-influenza drugs.

Keywords:  Anti-influenza; Enzyme hydrolysis; Kelp polysaccharide; Neuraminidase

DOI:  https://doi.org/10.1016/j.ijbiomac.2024.135506
J Chem Inf Model. 2024 Sep 09.

Developing and Benchmarking Sulfate and Sulfamate Force Field Parameters via Ab Initio Molecular Dynamics Simulations To Accurately Model Glycosaminoglycan Electrostatic Interactions.

Miguel Riopedre-Fernandez, Vojtech Kostal, Tomas Martinek, Hector Martinez-Seara, Denys Biriukov.

Glycosaminoglycans (GAGs) are negatively charged polysaccharides found on cell surfaces, where they regulate transport pathways of foreign molecules toward the cell. The structural and functional diversity of GAGs is largely attributed to varied sulfation patterns along the polymer chains, which makes understanding their molecular recognition mechanisms crucial. Molecular dynamics (MD) simulations, thanks to their unmatched microscopic resolution, have the potential to be a reference tool for exploring the patterns responsible for biologically relevant interactions. However, the capability of molecular dynamics force fields used in biosimulations to accurately capture sulfation-specific interactions is not well established, partly due to the intrinsic properties of GAGs that pose challenges for most experimental techniques. In this work, we evaluate the performance of molecular dynamics force fields for sulfated GAGs by studying ion pairing of Ca2+ to sulfated moieties─N-methylsulfamate and methylsulfate─that resemble N- and O-sulfation found in GAGs, respectively. We tested available nonpolarizable (CHARMM36 and GLYCAM06) and explicitly polarizable (Drude and AMOEBA) force fields, and derived new implicitly polarizable models through charge scaling (prosECCo75 and GLYCAM-ECC75) that are consistent with our developed "charge-scaling" framework. The calcium-sulfamate/sulfate interaction free energy profiles obtained with the tested force fields were compared against reference ab initio molecular dynamics (AIMD) simulations, which serve as a robust alternative to experiments. AIMD simulations indicate that the preferential Ca2+ binding mode to sulfated GAG groups is solvent-shared pairing. Only our scaled-charge models agree satisfactorily with the AIMD data, while all other force fields exhibit poorer agreement, sometimes even qualitatively. Surprisingly, even explicitly polarizable force fields display a notable disagreement with the AIMD data, likely attributed to difficulties in their optimization and possible inherent limitations in depicting high-charge-density ion interactions accurately. Finally, the underperforming force fields lead to unrealistic aggregation of sulfated saccharides, which qualitatively disagrees with our understanding of the soft glycocalyx environment. Our results highlight the importance of accurately treating electronic polarization in MD simulations of sulfated GAGs and caution against over-reliance on currently available models without thorough validation and optimization.

DOI: https://doi.org/10.1021/acs.jcim.4c00981
Drug Metab Dispos. 2024 Sep 09. pii: DMD-AR-2024-001813. [Epub ahead of print]

The simultaneous inhibition of SLC6A19 and BCRP transporters leads to an increase of indoxyl sulfate (a uremic toxin) in plasma and kidney.

Qingping Wang, Bogdan Munteanu, Alexander Marker, Yongyi Luo, Constanze Holz, John L Kane, Theresa Kuntzweiler, Emma-Jane Poulton, Maja Sedic, Zaid Jayyosi, Jens Riedel, Jennifer Fretland.

  SLC6A19 inhibitors are being studied as therapeutic agents for Phenylketonuria. In this work, a potent SLC6A19 inhibitor (RA836) elevated rat kidney uremic toxin indoxyl sulfate (IDS) levels by intensity (arbitrary unit) of 13.7{plus minus}7.7 compared to vehicle 0.3{plus minus}0.1 (P=0.01) as determined by tissue mass spectrometry imaging (tMSI) analysis. We hypothesized that increased plasma and kidney levels of IDS could be caused by the simultaneous inhibition of both Slc6a19 and a kidney IDS transporter responsible for excretion of IDS into urine. To test this, we first confirmed the formation of IDS through tryptophan metabolism by feeding rats a Trp-free diet. Inhibiting Slc6a19 with RA836 led to increased IDS in these rats. Next, RA836 and its key metabolites were evaluated in vitro for inhibiting kidney transporters OAT1, OAT3 and BCRP. RA836 inhibits BCRP with an IC50 of 0.045 µM but shows no significant inhibition of OAT1 or OAT3. Finally, RA836 analogs with either potent or no inhibition of SLC6A19 and/or BCRP were synthesized and administered to rats fed a normal diet. Plasma and kidney samples were collected to quantify IDS using LC-MS. Neither a SLC6A19 inactive but potent BCRP inhibitor nor a SLC6A19 active but weak BCRP inhibitor raised IDS levels, while compounds inhibiting both transporters caused IDS accumulation in rat plasma and kidney, supporting the hypothesis that rat Bcrp contributes to the excretion of IDS. In summary, we identified that inhibiting Slc6a19 increases IDS formation, while simultaneously inhibiting Bcrp results in IDS accumulation in the kidney and plasma. Significance Statement This is the first publication to decipher the mechanism for accumulation of IDS (a uremic toxin) in rats via inhibition of both Slc6a19 and Bcrp. Specifically, inhibition of Slc6a19 in the GI track increases IDS formation and inhibition of Bcrp in the kidney blocks IDS excretion. Therefore, we should avoid inhibiting both SLC6A19 and BCRP simultaneously in humans to prevent accumulation of IDS, a known risk factor for cardiovascular disease, psychic anxiety, and mortality in chronic kidney disease patients.

Keywords:  Mass spectrometry (MS); drug development/discovery; drug metabolism; drug transport; kidney disease; toxicity; transporters

DOI:  https://doi.org/10.1124/dmd.124.001813
Cancer Res. 2024 Sep 09.

Cancer-Associated Fibroblasts Expressing Sulfatase 1 Facilitate VEGFA-Dependent Microenvironmental Remodeling to Support Colorectal Cancer.

Huijuan Wang, Jiaxin Chen, Xiaoyu Chen, Yingqiang Liu, Jiawei Wang, Qing Meng, Huogang Wang, Ying He, Yujia Song, Jingyun Li, Zhenyu Ju, Peng Xiao, Junbin Qian, Zhangfa Song.

Tumor stroma plays a critical role in fostering tumor progression and metastasis. Cancer-associated fibroblasts (CAFs) are a major component of the tumor stroma. Identifying the key molecular determinants for the pro-tumor properties of CAFs could enable the development of more effective treatment strategies. Herein, through analyses of single-cell sequencing data, we identified a population of CAFs expressing high levels of sulfatase 1 (SULF1), which was associated with poor prognosis in colorectal cancer (CRC) patients. CRC models using mice with conditional SULF1 knockout in fibroblasts revealed the tumor-supportive function of SULF1+ CAFs. Mechanistically, SULF1+ CAFs enhanced the release of vascular endothelial growth factor A (VEGFA) from heparan sulfate proteoglycan (HSPG). The increased bioavailability of VEGFA initiated the deposition of extracellular matrix (ECM) and enhanced angiogenesis. In addition, intestinal microbiota-produced butyrate suppressed SULF1 expression in CAFs through its HDAC inhibitory activity. The insufficient butyrate production in CRC patients increased the abundance of SULF1+ CAFs, thereby promoting tumor progression. Importantly, tumor growth inhibition by HDAC inhibition was dependent on SULF1 expression in CAFs, and CRC patients with more SULF1+ CAFs were more responsive to treatment with the HDAC inhibitor chidamide. Collectively, these findings unveil the critical role of SULF1+ CAFs in CRC and provide a strategy to stratify CRC patients for HDAC inhibitor treatment.

DOI: https://doi.org/10.1158/0008-5472.CAN-23-3987
Int J Environ Health Res. 2024 Sep 10. 1-10

Acrylamide exposure, sex hormones, and pubertal status in Japanese adolescents.

Chisato Nagata, Keiko Wada, Michiyo Yamakawa, Masaaki Sugino, Tomoka Mori, Jun Ueyama, Yoshio Sumoto.

  Acrylamide may affect sex hormone levels and the timing of sexual maturation. The present study cross-sectionally examined interrelationship between the urinary metabolite of acrylamide exposure, serum sex hormone levels, and pubertal status in 408 Japanese adolescents aged 13-14 years. Their caregivers completed a questionnaire concerning the health status of their children, including pubertal maturation, and the lifestyles of children and parents. Pubertal status was queried by the Pubertal Development Scale. A major metabolite of acrylamide, N-acetyl-S-(2-carbamoylethyl)-cysteine (AAMA) in first-void urine samples. In male students, urinary AAMA was significantly inversely associated with testosterone, puberty stage, and facial hair growth after controlling for covariates. Serum testosterone and DHEAS were significantly positively associated with puberty stage. In female students, urinary AAMA was not associated with puberty stage, the indices, or any measured hormones. The data suggest that exposure to acrylamide may impact the pubertal development of boys through the effects on testosterone level.

Keywords:  Acrylamide; adolescents; puberty; testosterone

DOI:  https://doi.org/10.1080/09603123.2024.2401578