bims-supasi 2025-02-23 papers

bims-supasi

Biomed News

on Sulfation pathways and signalling

Issue of 2025–02–23
sixteen papers selected by
Jonathan Wolf Mueller, University of Birmingham

Utilizing 13C-Labeled Internal Standards to Advance the Analysis of Heparan Sulfate.
Effect of vgb gene on microbial chondroitin sulfate production in recombinant Escherichia coli pETM6-PACF-vgb and physicochemical characterization of produced chondroitin sulfate.
Serum amyloid A binding to glycosaminoglycans is synergistic with amyloid formation: Therapeutic targeting in the inflammation-linked amyloidosis.
A novel endo-1,3-fucanase in glycoside hydrolase family 187 provided a biotechnological tool for preparing sulfated fucan oligosaccharides.
Development of a method for simultaneous analysis of glycosaminoglycan disaccharides and evaluating the quality of chondroitin sulfate and hyaluronic acid in food raw materials.
Sustainable and biocompatible hybrid materials-based sulfated polysaccharides for biomedical applications: a review.
Presentation of CXCL12γ by heparan sulfate proteoglycans activates CXCR4 without desensitization in B cells.
Injectable Chondroitin Sulfate Microspheres with Gallic Acid-Magnesium MOF for Anti-Inflammatory and Cartilage Degeneration Alleviation in Osteoarthritis Treatment.
A CD25-CCR7 complex initiates non-canonical IL-2 signaling.
Unveiling the role of sex in the metabolism of indoxyl sulfate and apixaban.
Variant-Specific Interactions at the Plasma Membrane: Heparan Sulfate's Impact on SARS-CoV-2 Binding Kinetics.
Microneedle Assisted Melittin-Chondroitin Sulfate Administration for the Transdermal Therapy of Rheumatoid Arthritis.
Cationic Liposomes as Broad-spectrum Antidotes for Heparin-based Anticoagulants.
Heparan sulfate regulates myofibroblast heterogeneity and function to mediate niche homeostasis during alveolar morphogenesis.
Rosmarinic acid-chondroitin sulfate nanoconjugate for targeted treatment of ulcerative colitis.
Validation of a quantitation method for conjugated quercetin in human plasma.

Am J Physiol Cell Physiol. 2025 Feb 19.

Utilizing 13C-Labeled Internal Standards to Advance the Analysis of Heparan Sulfate.

Zhangjie Wang, Jian Liu.

  Heparan sulfate is a highly sulfated and structurally heterogeneous polysaccharide that plays key roles in numerous biological processes. Due to its complex structure and variable sulfation patterns, accurately characterizing and quantifying HS in biological samples poses significant analytical challenges. This review presents an advanced LC-MS/MS methodology that utilizes isotope-labeled internal standards for the precise quantification of HS disaccharides and rare 3-O-sulfated tetrasaccharides, alongside monitoring 6-O-endosulfatase enzyme activity and the metabolism of synthetic HS oligosaccharides in biological systems. The combination of isotope-labeled standards with LC-MS/MS technology provides a powerful and sensitive approach for comprehensive analysis of HS modifications, offering valuable insights into HS metabolism and its alterations across various biological contexts.

Keywords:  13C-labeled internal standards; 6-O-Sulfatase; HS disaccharide/ 3-O-sulfated tetrasaccharide; HS metabolism; Heparan sulfate; LC-MS/MS

DOI:  https://doi.org/10.1152/ajpcell.00944.2024
Cell Mol Biol (Noisy-le-grand). 2025 Feb 20. 71(2): 21-27

Effect of vgb gene on microbial chondroitin sulfate production in recombinant Escherichia coli pETM6-PACF-vgb and physicochemical characterization of produced chondroitin sulfate.

Ayse Sebnem Erenler, Tuba Unver, Ahmet Faruk Ceylan, Imren Ozcan, Hikmet Geckil.

Chondroitin Sulfate (CS) is an essential component of the extracellular matrix and is a sulfated glycosaminoglycan structurally composed of a polysaccharide chain consisting of N-acetyl galactosamine and glucuronic acid. The use of CS of animal origin is common in pharmacological research. The disadvantages of traditional sources and methods used in the production of CS, which is used in various applications in the medicine, veterinary, pharmacy, and cosmetic sectors, have made microbial production a vital alternative. In this study, recombinant Escherichia coli (pETM6-PACF-vgb) strain, in which kfoA, kfoC, kfoF and vgb gene regions are co-expressed, and E. coli pETM6-PACF strain, which does not contain the vgb gene, were used in the microbial production of CS. The vgb gene is the region responsible for expressing the bacterial protein Vitreoscilla hemoglobin (VtHb). This study investigated the effect of the expression of VtHb in E. coli on increasing bacterial cell respiration and, therefore, how ATP production would affect cell growth and the acquisition of chondroitin and microbial chondroitin sulfate (MCS) from biomass. The analysis results determined a 23.07% increase in the amount of MCS produced from the vgb+ strain. The presence of vgb had positively affected culture age and reproductive kinetics. Spectrophotometric measurements, NMR, HPLC, FT-IR, TGA, DTA, and DSC analyses for the reproductive values and physicochemical characterization of the obtained MCS were applied to discuss this production process. For more detailed results on this subject, future research focused on optimization is needed.

DOI: https://doi.org/10.14715/cmb/2025.71.2.4
J Mol Biol. 2025 Feb 13. pii: S0022-2836(25)00073-7. [Epub ahead of print] 169007

Serum amyloid A binding to glycosaminoglycans is synergistic with amyloid formation: Therapeutic targeting in the inflammation-linked amyloidosis.

Shobini Jayaraman, Angela Urdaneta, Marcus Fandrich, Olga Gursky.

  Serum amyloid A (SAA), a small lipophilic plasma protein elevated in inflammation, is a precursor of amyloid A (AA) amyloidosis, the major life-threatening complication of chronic inflammation in animals and humans. Although heparan sulfate (HS) is a potent amyloid agonist, particularly in AA amyloidosis, therapeutic targeting of SAA-HS interactions using a small-molecule HS/heparin decoy was unsuccessful. To understand molecular underpinnings, we used recombinant lipid-free human and murine SAA1 and human SAA2 to explore their interactions with various glycosaminoglycans at pH 5.5-7.4 during amyloid formation, from native protein to amyloid oligomers and fibrils. Effects of pH and glycosaminoglycan sulfation/charge supported by prior computational studies indicate electrostatic origin of SAA-glycosaminoglycan interactions. HS/heparin can promote amyloidogenesis by inducing non-native β -sheet and apparently causing liquid droplet formation in SAA in solution. Structural and binding studies by spectroscopy and ELISA reveal previously unknown synergy between amyloid formation and heparin/HS binding by SAA. We propose that this synergy potentially extends to other protein amyloids and stems from longitudinal binding of HS polyanions to basic residue arrays on amyloid oligomers or fibrils. This binding mode explains our finding that a minimal heparin chain length exceeding 20 monosaccharides is necessary to compete with HS for binding to amyloid oligomers. The results help explain prior failure of a small-molecule drug in targeting of SAA-HS interactions and consider alternative HS-targeting approaches for AA and, potentially, other amyloid diseases.

Keywords:  AA amyloidosis; electrostatic interactions; liquid droplets; short-chain and small-molecule heparin mimetics; sulfated glycosaminoglycans

DOI:  https://doi.org/10.1016/j.jmb.2025.169007
Int J Biol Macromol. 2025 Feb 16. pii: S0141-8130(25)01720-9. [Epub ahead of print] 141171

A novel endo-1,3-fucanase in glycoside hydrolase family 187 provided a biotechnological tool for preparing sulfated fucan oligosaccharides.

Jingjing Shen, Long Zheng, Guangning Chen, Achiraya Khamleng, Changhu Xue, Yaoguang Chang.

  Sulfated fucan, an important marine polysaccharide frequently presented in echinoderms and brown algae, has gained growing attention owing to its various biological activities. Fucanases are essential tools for degrading sulfated fucan to produce corresponding oligosaccharides. In this context, an endo-1,3-fucanase (Fun187Al) belonging to the GH187 family was successfully expressed in Escherichia coli. Fun187Al showed the highest activity at temperatures of 30-40 °C and pH 7.5. It hydrolyzed sulfated fucans in a random endo-acting pattern, and displayed a substrate specificity different from the endo-1,3-fucanases of other glycoside hydrolase family. Analyses of ultra-performance liquid chromatography coupled with high-resolution mass spectrometry revealed that tetrasaccharide with two sulfate groups (Fuc4S2), Fuc4S3, and Fuc4S4 were respectively the major components in the end products of Fun187Al against sulfated fucans from Acaudina molpadioides, Thelonota ananas, and Holothuria tubulosa. The capability of Fun187Al to produce oligosaccharides with different degrees of polymerization and sulfation patterns demonstrated that it could be regarded as a favorable tool for establishing the structure-activity relationship of sulfated fucan and its oligosaccharides.

Keywords:  Endo-1,3-fucanase; GH187; Sulfated fucan

DOI:  https://doi.org/10.1016/j.ijbiomac.2025.141171
Food Chem X. 2025 Jan;25 102239

Development of a method for simultaneous analysis of glycosaminoglycan disaccharides and evaluating the quality of chondroitin sulfate and hyaluronic acid in food raw materials.

Kengo Hirai, Takamasa Ishii, Ayaka Aijima, Nae Yokota, Yasuhisa Miyamoto, Kyohei Higashi, Yusuke Iwasaki, Rie Ito, Nobuaki Higashi, Hiroshi Akiyama.

  We developed a comprehensive liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for the simultaneous analysis of 19 glycosaminoglycan disaccharide components. Although structural similarity among components can hinder MS identification, we optimized the LC conditions to successfully separate all components. Specifically, we quantitatively and qualitatively evaluated chondroitin sulfate (CS) and hyaluronic acid (HA) in food ingredients. The method involves enzymatic degradation of CS and HA by Chondroitinase ABC into their constituent disaccharides, followed by LC-MS/MS quantification. We identified conditions that ensure optimal enzymatic digestion of CS and HA. Calibration curves showed excellent precision, accuracy, and linearity in validation tests. Quantitative analyses of CS and HA in commercially available raw materials were consistent with labeled purity. Disaccharide composition analysis suggested that the origin stated on the label was accurate. The findings suggest that the proposed method is useful for the quantitative and qualitative evaluation of the functional food ingredients CS and HA.

Keywords:  Chondroitin sulfate; Enzymatic digestion; Glycosaminoglycan; Hyaluronic acid; LC-MS/MS; Unsaturated disaccharide

DOI:  https://doi.org/10.1016/j.fochx.2025.102239
RSC Adv. 2025 Feb 06. 15(6): 4708-4767

Sustainable and biocompatible hybrid materials-based sulfated polysaccharides for biomedical applications: a review.

Reem S Alfinaikh, Khalid A Alamry, Mahmoud A Hussein.

Sustainable biomaterials that are both efficient and environmentally friendly are the subject of research and development efforts among scientists and academics from a variety of contemporary scientific disciplines. Due to their significant involvement in several physiological and pathological processes, sulfated polysaccharides (SPs) have garnered growing interest across various application domains, including biomedicine. Nevertheless, mechanical and thermal stability are issues for unmodified polysaccharide materials. Interactions between polymers, such as the mixing of biopolymers with synthetic or biopolymers through chemical interaction or grafting into the main chain structure of raw materials to enhance their therapeutic effects, are essential to meet the high standards of biomedical features. Another way to improve the mechanical and thermal properties is to graft appropriate fillers onto the polysaccharide backbone. The characteristics of polysaccharide bio-nanocomposites in comparison to more traditional polymers have attracted a lot of interest. With an emphasis on anti-inflammatory, anticancer, antiviral, immunoregulatory, and anticoagulant properties, this review delves into the most recent biological uses of sulfated polysaccharides. As well as thoroughly outlining the factors that impact the biological properties, such as the extraction process, molecular weight (Mw), the degree of sulfation, distribution/position, modification procedures, and the filler size, etc., this review aims to: (1) provide a systematic and critical overview of the cutting-edge research on SPs and hybrid sulfated polysaccharide bio-nanocomposites; (2) identify the key factors, mechanisms, methods, and challenges impacting SPs bio-nanocomposites; (3) elucidate the current and potential biomedical applications, advantages, manufacturing challenges, and opportunities associated with SPs bio-nanocomposites; (4) offer insights into future research directions by suggesting improvements for bio-nanocomposites, including novel materials, and advanced processing techniques.

DOI: https://doi.org/10.1039/d4ra07277d
Blood Adv. 2025 Feb 19. pii: bloodadvances.2024014396. [Epub ahead of print]

Presentation of CXCL12γ by heparan sulfate proteoglycans activates CXCR4 without desensitization in B cells.

Hildo C Lantermans, Fangxue Ma, Annemieke Kuil, Martin F M de Rooij, Nick Bergkamp, Werner Jack van der Meer, Jaap Diederik van Buul, Martine Smit, Marie José Kersten, Marcel Spaargaren, Steven T Pals.

CXCL12-CXCR4 signaling is involved in a wide variety of homeostatic and pathological processes, but the role of specific CXCL12 isoforms has remained largely unexplored. We have recently shown that the CXCL12γ isoform, which holds an exceptionally high affinity for heparan sulfate (HS), is produced by human bone marrow stromal cells (BMSCs) and remains cell-surface immobilized by HS proteoglycans (HSPGs). This HS-bound CXCL12γ is critical for the adhesion of multiple myeloma cells to BMSCs and for BMSC-mediated drug resistance. Here, we investigated how CXCL12γ activates and regulates CXCR4, by employing a variety of biosensors in HEK293T cells and endogenous CXCR4-expressing B-lymphoma and myeloma cell lines as well as primary B cells. We show that CXCL12γ and CXCL12α bind CXCR4 with a similar affinity and that the cumulative activation of CXCR4 over time is equal for both ligands, although CXCL12α activates CXCR4 more rapidly. Whereas non-bound CXCL12γ and CXCL12α equally induce CXCR4 internalization, cell- or heparin-bound CXCL12γ hardly induces CXCR4 internalization or desensitization. CXCL12γ presented by HSPGs on the membrane of human bone marrow endothelial cells (HBMECs) induces potent cell adhesion to the endothelium under physiological flow, while, upon encountering HBMEC-bound CXCL12γ, cells retain the ability to migrate towards CXCL12α. Taken together, our data demonstrate that CXCL12γ and CXCL12α differentially modulate CXCR4 trafficking and that CXCL12γ, immobilized and presented by HSPGs on the cell surface of HBMECs, can efficiently arrest circulating cells without causing CXCR4 internalization or desensitization, thus allowing subsequent cell migration towards a CXCL12α gradient.

DOI: https://doi.org/10.1182/bloodadvances.2024014396
ACS Appl Mater Interfaces. 2025 Feb 17.

Injectable Chondroitin Sulfate Microspheres with Gallic Acid-Magnesium MOF for Anti-Inflammatory and Cartilage Degeneration Alleviation in Osteoarthritis Treatment.

Jiachen He, Jianjun Wu, Jingcheng Zheng, Yidan Xu, Keyun Li, Siwei Yin, Yanyun Liu, Yuelin Hu, Chaoming Xie, Limin Cai, Yikuan Du, Xiong Lu.

  Inflammation and cartilage degeneration are critical challenges in osteoarthritis (OA) treatment. Achieving sustained drug efficacy while mitigating the adverse effects of inflammation and reactive oxygen species remains a significant challenge. This study synthesizes a gallic acid-magnesium (GA-Mg) metal-organic framework (MOF) as a drug carrier for puerarin (PA). The PA-loaded GA-Mg MOF (pGM) is encapsulated within chondroitin sulfate methacrylate, forming monodisperse hybrid microspheres (CM@pGM) under ultraviolet light using microfluidic technology. The pGM is physically confined within the microspheres through a network of structural obstructions and noncovalent interactions. During degradation, GA and Mg2+ ions release from pGM, improving the inflammatory microenvironment of the articular cavity and mitigating oxidative stress. The sustained release of Mg2+ and PA supports chondrocyte anabolism and facilitates cartilage repair. In vitro studies confirm that injectable microspheres extend the drug release period to over 2 weeks. In vivo experiments demonstrate that CM@pGM significantly reduces osteophyte formation, alleviates degenerative changes in articular cartilage, and delays OA progression. In conclusion, CM@pGM, as a drug delivery platform that ameliorates the inflammatory microenvironment, alleviates oxidative stress, and promotes cartilage repair, holds significant potential for OA treatment.

Keywords:  anti-inflammation; cartilage degeneration; metal−organic frameworks; microspheres; osteoarthritis

DOI:  https://doi.org/10.1021/acsami.4c22415
bioRxiv. 2025 Feb 04. pii: 2025.02.03.636356. [Epub ahead of print]

A CD25-CCR7 complex initiates non-canonical IL-2 signaling.

Sarah Hyun Ji Kim, Hosup Lee, Alexandre Gingras, Klaus Ley, Jamie B Spangler, Mark H Ginsberg.

IL-2, a central regulator of immune function, binds to its receptor subunit CD25 (IL-2Rα), promoting IL-2 interaction with β and γ subunits to trigger the canonical IL-2 signaling pathway. An anti-mouse CD25 antibody, PC61, triggers alternative IL-2 signaling, leading to integrin activation. PC61 induces a complex formed by the IL-2-dependent association of CD25 with CCR7, suggesting that the formation of this complex initiates alternative IL-2 signaling. Here, we used structure-based design together with combinatorial screening to identify an IL-2 mutant (denoted IL-2(E52K)) that spares canonical IL-2 signaling but disrupts both PC61-induced complex formation and integrin activation while retaining the full CD25 affinity of the parent molecule. We also report that heparan sulfate (HS), a physiological ligand of IL-2 that triggers alternative signaling, induced IL-2-dependent CD25-CCR7 association, whereas IL-2(E52K) failed to support both HS-induced CD25-CCR7 complex formation and integrin activation. Thus, both anti-CD25 antibody and HS require common features of IL-2 needed for CD25-CCR7 complex assembly and resulting integrin activation. Collectively, these data show that IL-2 promotes CD25 interaction with CCR7, thereby forming the signal initiating complex. Furthermore, canonical and alternative IL-2 signaling can be decoupled by an IL-2 mutation, creating a tool to specify the biological role of alternative IL-2 signaling in immune responses.

DOI: https://doi.org/10.1101/2025.02.03.636356
Sci Rep. 2025 Feb 19. 15(1): 6075

Unveiling the role of sex in the metabolism of indoxyl sulfate and apixaban.

Blanca Pina-Beltran, Daniel Dimitrov, Nathalie McKay, Matthieu Giot, Zbyněk Zdráhal, David Potěšil, Václav Pustka, Jorge Peinado-Izaguerri, Julio Saez-Rodriguez, Stéphane Poitevin, Stéphane Burtey.

Chronic Kidney Disease (CKD) is associated with heightened risk of thrombosis. Prescription of anticoagulants is key to manage it; however, CKD patients have shown an increased risk of bleeding under anticoagulation therapy compared to non-CKD patients. We hypothesized that the sex could modify the metabolism of indoxyl sulfate (IS), a uremic toxin and Apixaban. Our intoxication model shows that higher doses of IS and apixaban accumulate in the plasma of female mice because of expression differences in efflux transporters and cytochromes in the liver, ileum and kidneys, when compared to males. Furthermore, we found that accumulation of apixaban in females contributes to increased bleeding. Transcriptional analysis of liver samples revealed elevated Sult1a1 but reduced Abcg2 and Cyp3a11 in female mice, while in the kidneys the expression rates of Oat1 and Oat3 were respectively lower and higher than those observed in males, potentially affecting drug clearance. Whole proteomics liver analysis confirmed the previous transcriptional results at the protein level and revealed that sex had a major influence in regulating both coagulation and drug metabolism pathways. Thus, our findings underline the need for inclusive clinical and preclinical trials to accurately reflect sex-specific metabolic variations, and to consider CKD-specific changes to optimize dosing, minimize side effects, and improve patient outcomes.

DOI: https://doi.org/10.1038/s41598-025-90405-5
Anal Chem. 2025 Feb 20.

Variant-Specific Interactions at the Plasma Membrane: Heparan Sulfate's Impact on SARS-CoV-2 Binding Kinetics.

Dario Valter Conca, Fouzia Bano, Małgorzata Graul, Julius von Wirén, Lauriane Scherrer, Hudson Pace, Himanshu Sharma, Justas Svirelis, Konrad Thorsteinsson, Andreas Dahlin, Marta Bally.

The spread of SARS-CoV-2 led to the emergence of several variants of concern (VOCs). The spike glycoprotein, responsible for engaging the viral receptor, exhibits the highest density of mutations, suggesting an ongoing evolution to optimize viral entry. This study characterizes the bond formed by virion mimics carrying the SARS-CoV-2 spike protein and the plasma membrane of host cells in the early stages of virus entry. Contrary to the traditional analysis of isolated ligand-receptor pairs, we utilized well-defined biomimetic models and biochemical and biophysical techniques to characterize the multivalent interaction of VOCs with the complex cell membrane. We observed an overall increase in the binding affinity for newer VOCs. By progressively reducing the system complexity, we identify heparan sulfate (HS) as a main driver of this variation, with a 10-fold increase in affinity for Omicron BA.1 over that of the original strain. These results demonstrate the essential role of coreceptors, particularly HS, in the modulation of SARS-CoV-2 infection and highlight the importance of multiscale biophysical and biochemical assays that account for membrane complexity to fully characterize and understand the role of molecular components and their synergy in viral attachment and entry.

DOI: https://doi.org/10.1021/acs.analchem.4c04283
Adv Healthc Mater. 2025 Feb 19. e2400543

Microneedle Assisted Melittin-Chondroitin Sulfate Administration for the Transdermal Therapy of Rheumatoid Arthritis.

Lili Jin, Zhenhui Wang, Cheng Peng, Miao He, Feng Wang, Hongyan He, Changsheng Liu.

  Rheumatoid arthritis (RA), a persistent and debilitating chronic disease characterize by joint damage and deformity, significantly impairs the life quality of patients and presents challenges for conventional drug administration due to organ damage and unsatisfactory therapeutic outcomes. To address these challenges, this study introduces an innovative hydrogel cryo-microneedle patch (CMNP)-mediated local administration system, primarily composed of chondroitin sulfate (CS), to deliver the potent anti-inflammatory drug melittin (MEL). This innovative approach not only circumvents organ impairment but also enhances patient compliance. The acute toxicity of MEL is effectively mitigated by electrostatic binding with CS molecules, forming MC complexes that induce apoptosis in fibroblast-like synoviocytes (FLS). The MC-loaded CMNPs (MC@CCMNPs) exhibit remarkable therapeutic capacity with a notably meliorated joint damage and suppressed arthritis severity in the RA rat model. Therefore, MC@CCMNPs emerge as a promising anti-inflammatory and safe therapy for RA treatment, as well as for other inflammation related chronic diseases.

Keywords:  chondroitin sulphate; melittin; microneedles; rheumatoid arthritis; surface‐enhanced Raman scattering

DOI:  https://doi.org/10.1002/adhm.202400543
Acta Biomater. 2025 Feb 19. pii: S1742-7061(25)00131-X. [Epub ahead of print]

Cationic Liposomes as Broad-spectrum Antidotes for Heparin-based Anticoagulants.

Jiarui Du, Yang Liu, Zhenzhen Dong, Yubiao Huang, Yang An, Xiaoyu Cheng, Ge Sun, Chong Du, Guangjun Nie, Xueqin Hou, Yinlong Zhang.

  Heparin-based anticoagulants have been widely used for the prevention and treatment of venous thrombotic diseases, as well as for anticoagulation during cardiopulmonary bypass and hemodialysis. However, excessive heparin usage brings serious bleeding risk, necessitating immediate reversal of their anticoagulant activity. Additionally, to prevent bleeding during surgery and restore hemostatic function post-cardiopulmonary bypass and hemodialysis, it is also crucial to reverse heparin's anticoagulant effects. Currently, protamine sulfate (PS) is the only clinically approved antidote for heparin. However, its effectiveness against low molecular weight heparin (LMWH) and fondaparinux sodium is limited. Moreover, PS has great potential to trigger fatal allergic reactions. Despite these concerns, no successful clinical substitutes for PS have been developed. In the current work, drawing inspiration from the mechanism by which PS efficiently reverses heparin, we modified the cationic liposome with cationic amino acids, arginine and lysine, to serve as a broad-spectrum antidote (CRKRK-Lipo) for heparin-based anticoagulants. This modification not only enhances their reversal efficiency but also reduces the overall surface charge, potentially improving their biocompatibility. In the tail bleeding and liver injury mouse models, CRKRK-Lipo demonstrated reversal efficiency comparable to PS for heparin and superior reversal efficiency for LMWH and fondaparinux sodium. Notably, CRKRK-Lipo exhibited a wider therapeutic dose window and did not exhibit severe cytotoxicity or immunogenicity, in contrast to PS. It is worth noting that cationic liposomes without polypeptide modification also displayed a significant heparin reversal effect. Our findings not only offer a potential alternative for PS but also broaden the application fields of cationic liposome. STATEMENT OF SIGNIFICANCE: This study introduces the cationic liposomes as a novel and effective alternative to protamine sulfate (PS) for the functional reversal of heparin-based anticoagulants. The results reveal that both CRKRK-modified cationic liposomes (CRKRK-Lipo) and unmodified cationic liposomes (Lipo) showed comparable reversal efficiency to PS for UFH and superior reversal efficiency for LMWH and fondaparinux sodium, with a wider therapeutic dose window and reduced toxicity. This work offers an alternative strategy for detoxifying heparin-based anticoagulants and expands the biomedical applications of cationic liposomes.

Keywords:  Cationic liposomes; Heparin-based anticoagulants; Immunogenicity; Protamine sulfate (PS); Reversal agents

DOI:  https://doi.org/10.1016/j.actbio.2025.02.037
Nat Commun. 2025 Feb 21. 16(1): 1834

Heparan sulfate regulates myofibroblast heterogeneity and function to mediate niche homeostasis during alveolar morphogenesis.

Hua He, Chong Ma, Wei Wei, Haonan Wang, Yutian Lai, Ming Liu, Shenfei Sun, Qing Ma, Jiashuang Lai, Hanxiang Liu, Hanmin Liu, Fei Sun, Xinhua Lin.

Postnatal respiration requires bulk formation of alveoli that produces extensive surface area for gas diffusion from epithelium to the circulatory system. Alveolar morphogenesis initiates at late gestation or postnatal stage during mammalian development and is mediated by coordination among multiple cell types. Here we show that fibroblast-derived Heparan Sulfate Glycosaminoglycan (HS-GAG) is essential for maintaining a niche that supports alveolar formation by modulating both biophysical and biochemical cues. Gli1-CreER mediated deletion of HS synthase gene Ext1 in lung fibroblasts results in enlarged and simplified alveolar structures. Ablation of HS results in loss of a subset of PDGFRαhi αSMA+ alveolar myofibroblasts residing in the distal alveolar region, which exhibit contractile properties and maintain WNT signaling activity to support normal proliferation and differentiation of alveolar epithelial cells. HS is essential for proliferation while preventing precocious apoptosis of alveolar myofibroblasts. We show that these processes are dependent upon FGF/MAPK signaling and forced activation of MAPK/ERK signaling partially corrected alveolar simplification and restored alveolar myofibroblast number and AT2 cell proliferation in HS deficient mice. These data reveal HS-dependent myofibroblast heterogeneity and function as an essential orchestrator for developing alveolar niche critical for the generation of gas exchange units.

DOI: https://doi.org/10.1038/s41467-025-57163-4
Int J Biol Macromol. 2025 Feb 17. pii: S0141-8130(25)01557-0. [Epub ahead of print] 141008

Rosmarinic acid-chondroitin sulfate nanoconjugate for targeted treatment of ulcerative colitis.

Miaomiao Long, Jie Li, Meiyang Yang, Weijun Chen, Lipeng Qiu, Xian Cheng, Liangwu Bi.

  Rosmarinic acid (RA) is an attractive candidate for ulcerative colitis (UC) application due to its bioactive properties, including antioxidant and anti-inflammatory functions, however, the poor water solubility and on-targeting hamper its therapeutic outcome. Therefore, this work reported the synthesis and preparation of novel water-soluble rosmarinic acid-chondroitin sulfate A (RA-CSA) nanoconjugate, which was used for the treatment of UC in dextran sulfate sodium (DSS)-induced acute colitis mouse model. RA was functionalized with CSA as confirmed by FTIR and 1H NMR, and self-assembled to form nanoassemblies with a diameter of 247.3 ± 2.99 nm. RA-CSA nanoassemblies exhibited radical scavenging and antioxidant capacity. RA-CSA remarkably inhibited lipopolysaccharide-induced nitric oxide and TNF-α production in RAW 264.7 cells without cytotoxicity, whose inhibition rate was <5 % at 200 μg mL-1. Oral administration of RA-CSA nanoassemblies significantly attenuated colonic inflammation compared to the parent RA, as evidenced by significantly reduced the shortening of colon length (4.20 ± 0.15 cm), body weight loss, and colonic inflammatory damage in DSS-induced colitis mice. In addition, RA-CSA nanoassemblies suppressed the expression and production of typical pro-inflammatory cytokines of ulcerative colitis. These results suggest that RA-CSA nanoassemblies deserve further consideration as a potential therapeutic drug for the treatment of UC.

Keywords:  Chondroitin sulfate; Rosmarinic acid; Ulcerative colitis

DOI:  https://doi.org/10.1016/j.ijbiomac.2025.141008
J Pharm Biomed Anal. 2025 Feb 12. pii: S0731-7085(25)00079-2. [Epub ahead of print]258 116738

Validation of a quantitation method for conjugated quercetin in human plasma.

Yui Sudaka, Takafumi Mitsui, Hiroaki Kida, Mst Julia Sultana, Miyu Nishikawa, Shinichi Ikushiro, Naoto Yamaguchi.

  Since the type of glycoside affects the pharmacokinetic profile of the aglycon after oral ingestion of quercetin glycosides, clinical studies on the pharmacokinetics of quercetin glycosides are required. However, a suitable method to determine the concentrations of quercetin phase II metabolites in human plasma and urine is lacking. Therefore, we developed and validated an LC-MS method for the quantitation of conjugated quercetin using relevant reference standards, including hetero-conjugates with glucuronic acid and sulfonic acid (QC-GA/S). Quercetin hetero-conjugates extracted from rat serum were used for the method development, and reference standards were biosynthesized for the quantitation. The use of a solid-phase extraction (SPE) column in a 96 well format enabled high-throughput analysis of up to 96 tests in a day, without compromising recovery and sensitivity. The SPE column with a weak anion exchange group contributed to the high recovery of QC-GA/S. The method was then validated, and its usefulness was confirmed using clinical samples. QC-GA/S was the predominant phase II quercetin metabolite after the ingestion of quercetin glucoside or quercetin supplements. Moreover, the two peaks of QC-GA/S found in human plasma and urine were isomers of QC-7GA/4'S, which has been reported as the predominant peak in rat plasma. If QC-GA/S in plasma is responsible for a physiological activity of quercetin, it is important to determine the concentration of each QC-GA/S isomer.

Keywords:  Flavonoids; LC-MS; Pharmacokinetics; Phase-II-metabolites; Quercetin; Solid-phase-extraction

DOI:  https://doi.org/10.1016/j.jpba.2025.116738