RESUMEN
Osteoclast fusion is a critical step for bone resorption, and its dysregulation contributes to pathological bone loss such as postmenopausal osteoporosis. Siglec15, a membrane-bound lectin, has emerged as a key regulator of osteoclast precursor recognition and fusion. Herein, it is reported that Cajanin, a natural small-molecule compound, potently inhibits osteoclastogenesis and bone resorption both in vitro and in vivo. Cajanin treatment suppressed actin ring formation, bone resorptive activity and the expression of osteoclastic markers including Ctsk and Mmp9 in a dose-dependent manner. Transcriptomic profiling revealed that Cajanin downregulated key fusion mediators such as Siglec15, DC-STAMP, OC-STAMP and NFATc1. Gene enrichment analysis identified attenuated MAPK and NF-κB signaling, which was validated by decreased phosphorylation of ERK, JNK and p38, increased IκB-α levels and reduced nuclear translocation of p65. Moreover, Cajanin significantly suppressed NFATc1 expression and nuclear localization. In an ovariectomy-induced bone loss model, Cajanin preserved trabecular bone architecture and reduced TRAP-positive osteoclasts. Collectively, these findings demonstrate that Cajanin transcriptionally disrupts the Siglec15-NFATc1 signaling axis, thereby impairing osteoclast fusion and bone resorption, and highlight its potential as a therapeutic candidate for selectively targeting pathological osteoclast fusion in osteolytic diseases.
Asunto(s)
Resorción Ósea , Lectinas , Factores de Transcripción NFATC , Osteoclastos , Animales , Factores de Transcripción NFATC/metabolismo , Factores de Transcripción NFATC/genética , Osteoclastos/efectos de los fármacos , Osteoclastos/fisiología , Osteoclastos/metabolismo , Resorción Ósea/tratamiento farmacológico , Resorción Ósea/metabolismo , Transducción de Señal/efectos de los fármacos , Femenino , Ratones , Osteogénesis/efectos de los fármacos , Ratones Endogámicos C57BL , Humanos , Lectinas/genética , Lectinas/metabolismo , Células RAW 264.7 , OvariectomíaRESUMEN
Hypertension, a major risk factor for cardiovascular diseases, is closely associated with excessive sodium intake and affects millions globally. Omentin-1, an adipokine with anti-inflammatory and antioxidant properties, has been implicated in blood pressure regulation, but its role in salt-sensitive hypertension remains unclear. This study investigated the antihypertensive and renal protective effects of omentin-1 in a deoxycorticosterone acetate (DOCA)-salt hypertensive rat model. We demonstrated that Omentin-1 overexpression significantly reduced blood pressure and attenuated renal dysfunction in DOCA-salt hypertensive rats, as evidenced by decreased serum creatinine (Scr) and blood urea nitrogen (BUN) levels, reduced renal fibrosis, and improved histopathological scores. Mechanistically, Omentin-1 downregulated angiotensin II type 1 receptor (AT1R) expression in the kidney, which was associated with reduced G protein-coupled receptor kinase 4 (GRK4) levels. Further analysis revealed that omentin-1 suppressed GRK4 expression via the reactive oxygen species (ROS)/c-Myc signaling pathway. Overexpression of GRK4 abolished the antihypertensive and renal protective effects of omentin-1, confirming GRK4 as a key mediator in this process. These findings highlight omentin-1 as a potential therapeutic target for salt-sensitive hypertension, offering dual benefits in blood pressure reduction and renal protection through the ROS/c-Myc/GRK4/AT1R axis.
Asunto(s)
Citocinas , Quinasa 4 del Receptor Acoplado a Proteína-G , Hipertensión , Lectinas , Proteínas Proto-Oncogénicas c-myc , Especies Reactivas de Oxígeno , Receptor de Angiotensina Tipo 1 , Cloruro de Sodio Dietético , Animales , Hipertensión/metabolismo , Hipertensión/inducido químicamente , Hipertensión/prevención & control , Hipertensión/tratamiento farmacológico , Especies Reactivas de Oxígeno/metabolismo , Masculino , Lectinas/genética , Lectinas/metabolismo , Ratas , Citocinas/genética , Citocinas/metabolismo , Regulación hacia Abajo/fisiología , Regulación hacia Abajo/efectos de los fármacos , Receptor de Angiotensina Tipo 1/metabolismo , Receptor de Angiotensina Tipo 1/genética , Transducción de Señal/fisiología , Transducción de Señal/efectos de los fármacos , Quinasa 4 del Receptor Acoplado a Proteína-G/metabolismo , Quinasa 4 del Receptor Acoplado a Proteína-G/genética , Proteínas Proto-Oncogénicas c-myc/metabolismo , Proteínas Ligadas a GPI/genética , Ratas Sprague-Dawley , Acetato de DesoxicorticosteronaRESUMEN
Background: Alpha-fetoprotein (AFP) and its isoform AFP-L3 are well-established serum biomarkers for hepatocellular carcinoma (HCC), a common malignancy and a leading cause of cancer-related mortality worldwide. Current methods for measuring these biomarkers are primarily lectin-based assays including the liquid-phase binding assay (LiBA) and liquid chromatography-tandem mass spectrometry (LC-MS/MS), both of which have limitations in diagnostic sensitivity and clinical utility for samples with low AFP concentrations. We aimed to develop a lectin-independent LC-MS/MS method for quantifying fucosylated AFP proteins (AFP-Fuc%). Methods: We conducted analytical validation, including method comparisons, over 2 months. The analytical sensitivity and diagnostic performance of this method were evaluated using 525 human serum samples-235 from HCC patients and 290 from non-HCC individuals-and compared with those of LiBA, which measured AFP-L3 levels. Results: The LC-MS/MS method demonstrated acceptable within-laboratory imprecision (CVs<17.1%) without detectable bias, carryover, or matrix effects. Our method exhibited a broader linear dynamic range (spanning five orders of magnitude) and 10-fold higher analytical sensitivity than LiBA. The diagnostic performance of our method was significantly superior to that of LiBA, particularly in patients with low AFP concentrations (<7 ng/mL, P <0.001), with improved accuracy, sensitivity, and precision at a specificity of 96.2%. Conclusions: The validated LC-MS/MS method demonstrated robust analytical performance and superior diagnostic accuracy over LiBA for HCC diagnosis while avoiding the inherent limitations of lectin-based assays. Our LC-MS/MS assay shows promise for early HCC detection and may contribute to enhanced patient care.
Asunto(s)
Espectrometría de Masas en Tándem , alfa-Fetoproteínas , Humanos , alfa-Fetoproteínas/análisis , alfa-Fetoproteínas/metabolismo , Espectrometría de Masas en Tándem/métodos , Cromatografía Liquida/métodos , Carcinoma Hepatocelular/diagnóstico , Carcinoma Hepatocelular/sangre , Neoplasias Hepáticas/diagnóstico , Neoplasias Hepáticas/sangre , Lectinas/metabolismo , Glicosilación , Biomarcadores de Tumor/sangre , Límite de Detección , Sensibilidad y Especificidad , FemeninoRESUMEN
Tumor-associated macrophages (TAM) in the pancreatic ductal adenocarcinoma (PDAC) tumor microenvironment (TME) exhibit immunosuppressive phenotypes and impaired phagocytic activity, facilitating tumor progression and immune evasion. In this study, we identified integrin α3ß1, composed of ITGA3 and ITGB1 subunits, as a sialylated glycoprotein ligand for Siglec-10, an inhibitory glyco-immune checkpoint receptor highly expressed on TAMs in PDAC. The interaction between Siglec-10 on TAMs and α3ß1 on PDAC cells suppressed macrophage-mediated phagocytosis, thereby promoting immune evasion. Consistently, disrupting Siglec-10 interactions using mAbs significantly enhanced macrophage phagocytosis of PDAC cells and alleviated myeloid cell-mediated inhibition of T-cell proliferation and activation in vitro. In both a xenograft mouse model engrafted with human macrophages and a human Siglec-10 transgenic mouse model, targeting Siglec-10 with mAbs reduced PDAC growth. These findings suggest that Siglec-10 interactions are key mediators of TAM-driven immune evasion in PDAC and highlight the therapeutic potential of targeting these interactions to restore antitumor immunity. SIGNIFICANCE: Pancreatic tumor cells exploit integrin α3ß1 to engage the immunosuppressive checkpoint receptor Siglec-10 on myeloid cells, driving immune evasion, which can be targeted with antibody-mediated blockade of Siglec-10 to restore antitumor immunity.
Asunto(s)
Carcinoma Ductal Pancreático , Integrina alfa3beta1 , Lectinas , Macrófagos , Neoplasias Pancreáticas , Fagocitosis , Macrófagos Asociados a Tumores , Animales , Humanos , Neoplasias Pancreáticas/inmunología , Neoplasias Pancreáticas/patología , Neoplasias Pancreáticas/metabolismo , Ratones , Fagocitosis/inmunología , Carcinoma Ductal Pancreático/inmunología , Carcinoma Ductal Pancreático/patología , Carcinoma Ductal Pancreático/metabolismo , Integrina alfa3beta1/metabolismo , Integrina alfa3beta1/inmunología , Microambiente Tumoral/inmunología , Macrófagos Asociados a Tumores/inmunología , Macrófagos Asociados a Tumores/metabolismo , Línea Celular Tumoral , Ensayos Antitumor por Modelo de Xenoinjerto , Ratones Transgénicos , Macrófagos/inmunología , Lectinas/metabolismo , Lectinas/inmunología , Anticuerpos Monoclonales/farmacologíaRESUMEN
The black tiger shrimp, Penaeus monodon, is cultivated commercially in many countries, including Thailand. However, diseases are major limiting factors in shrimp production. Understanding the shrimp immune responses to pathogens will be essential for health management and disease control. Fibrinogen-related proteins (FREPs) act as pattern recognition receptors by recognizing carbohydrate residues on pathogen surfaces. Here, we report that P. monodon FREP (PmFREP) interacts with the P. monodon lipopolysaccharide and ß-1,3-glucan binding protein (PmLGBP) to co-ordinate the innate immune response in shrimp. Like PmLGBP, PmFREP participates in the prophenoloxidase (proPO)-activating cascade upon the Vibrio parahaemolyticus acute hepatopancreatic necrosis disease-causing strain infection. PmFREP knockdown results in a significant decrease in phenoloxidase (PO) activity, whereas recombinant PmFREP injection noticeably increases the enzyme activity. The coiled-coil (CC) region at the N-terminus of PmFREP mediates the direct binding between PmFREP and PmLGBP to enhance PO activity. Our results suggest that PmFREP recognizes the pathogens to form an immunological complex with PmLGBP through the CC region, subsequently enhancing the proPO-activating cascade to eliminate the pathogens. Therefore, PmFREP serves as a key component for signaling transduction in the shrimp immune defense.
Asunto(s)
Proteínas de Artrópodos , Proteínas Portadoras , Inmunidad Innata , Lectinas , Lipopolisacáridos , Penaeidae , Animales , Penaeidae/inmunología , Penaeidae/microbiología , Penaeidae/metabolismo , Penaeidae/genética , Lipopolisacáridos/metabolismo , Lipopolisacáridos/inmunología , Proteínas Portadoras/metabolismo , Proteínas Portadoras/inmunología , Proteínas Portadoras/genética , Lectinas/metabolismo , Lectinas/inmunología , Lectinas/genética , Proteínas de Artrópodos/metabolismo , Proteínas de Artrópodos/genética , Proteínas de Artrópodos/inmunología , Vibrio parahaemolyticus/inmunología , Precursores Enzimáticos/metabolismo , Catecol Oxidasa/metabolismo , beta-Glucanos/metabolismo , Unión ProteicaRESUMEN
Glycans are essential membrane components that play critical roles in various biological processes, and their aberrant expression is often associated with diseases such as cancer. Lectins are carbohydrate binding proteins capable of binding specific glycan structures and have been widely used to study glycosylation patterns on cell membranes. However, multiplex glycan profiling of cancer-derived small extracellular vesicles (sEVs) using lectins remains limited due to their high heterogeneity, the small surface area of individual vesicles, the low abundance of specific glycan motifs, and the lack of robust multiplexing platforms. In this study, we developed a lectin-based surface enhanced Raman spectroscopy (SERS) assay for both individual glycan structure detection and simultaneous multiplex profiling of glycans on the surface of cancer cell-derived sEVs in a single test. We evaluated the sensitivity and specificity of this platform and validated its accuracy through a proof-of-concept study by detecting changes in the surface glycan profile of sEVs following glycosidase treatment. Using four different lectin-based SERS nanotags, we performed multiplex glycan profiling and observed distinct changes in glycan signatures after glycosidase treatment. These findings highlight the strong potential of this assay for multiplex sEV glycan profiling in a single sample with a single test, indicating its potential for future clinical applications, particularly in cancer diagnosis.
Asunto(s)
Vesículas Extracelulares , Lectinas , Polisacáridos , Espectrometría Raman , Espectrometría Raman/métodos , Vesículas Extracelulares/química , Vesículas Extracelulares/metabolismo , Polisacáridos/análisis , Polisacáridos/química , Polisacáridos/metabolismo , Humanos , Lectinas/química , Lectinas/metabolismo , Línea Celular TumoralRESUMEN
Bispecific antibodies and chimeric antigen receptor T cells are some of the most potent cancer immunotherapeutics in clinical use, yet most cancers remain poorly targetable. High-affinity antibodies required to maximize killing detect low antigen expression in normal tissue, risking "on-target, off-cancer" toxicity. This compels identification of cancer-restricted cell-surface protein antigens, which are rare. Tumor-associated carbohydrate antigens (TACAs) are the most abundant and widespread cancer antigens known but are poorly targetable by antibodies. Here, we describe glycan-dependent T cell recruiter (GlyTR) pan-cancer immunotherapeutics that utilize high-avidity "velcro-like" lectin binding to kill cells with high but not low TACA expression. GlyTR1 and GlyTR2 bind immunosuppressive ß1,6GlcNAc-branched N-glycans or multiple TACAs (Tn, sialyl-Tn, LacDiNAc, and GD2), respectively, overcome immunosuppressive mechanisms in the tumor microenvironment and trigger target-density-dependent T cell-mediated pan-cancer killing, yet they lack toxicity in mice with human-like TACA expression. Density-dependent lectin binding to TACAs provides highly potent and safe pan-cancer immunotherapeutics.
Asunto(s)
Antígenos de Carbohidratos Asociados a Tumores , Inmunoterapia , Neoplasias , Animales , Humanos , Antígenos de Carbohidratos Asociados a Tumores/inmunología , Antígenos de Carbohidratos Asociados a Tumores/metabolismo , Ratones , Inmunoterapia/métodos , Neoplasias/inmunología , Neoplasias/terapia , Polisacáridos/metabolismo , Polisacáridos/inmunología , Microambiente Tumoral/inmunología , Linfocitos T/inmunología , Línea Celular Tumoral , Femenino , Terapia de Inmunosupresión , Lectinas/metabolismoRESUMEN
More than 20 genes expressed in the male reproductive tract have been identified as essential factors for sperm migration to and through the utero-tubal junction (UTJ), and they are divided into ADAM3-dependent and ADAM3-independent pathways. In parallel, sperm having UTJ migration defects also show impaired binding to the zona pellucida (ZP). Herein, we demonstrate that knockout of Galntl5, encoding a sperm surface protein, causes impaired sperm binding with the UTJ and ZP, and null males have severe infertility. GALNTL5 appreciably disappears in sperm lacking Adam3 or Lypd4, required for ADAM3-dependent and ADAM3-independent pathways, and GALNTL5 binds to N-acetylgalactosamine (GalNAc) distributed on the UTJ and ZP. Blockage of GalNAc decreases the number of sperm binding to the UTJ and ZP. Thus, we unveil that GALNTL5 is a responsible factor for UTJ migration and sperm-ZP binding, and that sperm bind to the UTJ and ZP through interaction of GALNTL5 and GalNAc.
Asunto(s)
Acetilgalactosamina , Trompas Uterinas , Lectinas , N-Acetilgalactosaminiltransferasas , Interacciones Espermatozoide-Óvulo , Espermatozoides , Útero , Zona Pelúcida , Masculino , Zona Pelúcida/metabolismo , Animales , Espermatozoides/metabolismo , Femenino , Acetilgalactosamina/metabolismo , N-Acetilgalactosaminiltransferasas/metabolismo , N-Acetilgalactosaminiltransferasas/genética , Ratones , Ratones Noqueados , Trompas Uterinas/metabolismo , Interacciones Espermatozoide-Óvulo/fisiología , Motilidad Espermática , Útero/metabolismo , Unión Proteica , Lectinas/metabolismo , Lectinas/genética , Infertilidad Masculina/genética , Infertilidad Masculina/metabolismo , Polipéptido N-Acetilgalactosaminiltransferasa , Humanos , Glicoproteínas de Membrana , Proteínas ADAMRESUMEN
Gastric cancer remains a major global health burden, with limited response rates to current immunotherapies targeting the programmed death-ligand 1 (PD-1/PD-L1) axis. Recent studies have identified sialic acid-binding immunoglobulin-like lectin 15 (SIGLEC-15) as a novel immune checkpoint molecule that may drive immune evasion through PD-L1-independent pathways. This study aimed to evaluate the expression patterns of SIGLEC-15 and PD-L1 in gastric adenocarcinoma and to investigate their associations with clinicopathological features and patient outcomes. We retrospectively analyzed 133 consecutive cases of gastric adenocarcinoma with complete clinicopathologic and follow-up data. Immunohistochemical staining was performed on formalin-fixed tumor samples; SIGLEC-15 expression on tumor cells was quantified by H-score (high expression defined as ≥110) and PD-L1 status by combined positive score (CPS, positive if ≥1). High SIGLEC-15 expression correlated with multiple adverse pathological features, including lymphatic (p = 0.003), venous (p = 0.030), and perineural invasion (p = 0.010), and was associated with significantly poorer 3-year overall survival (hazard ratio = 3.36, p < 0.001). While SIGLEC-15 and PD-L1 expression were not mutually exclusive, an inverse relationship was generally observed. Patients with dual positivity (SIGLEC-15 high/PD-L1 CPS ≥ 1) showed the lowest 36-month survival (32%), compared to 56% in the dual-negative group (SIGLEC-15 low/PD-L1 CPS < 1). These results highlight the clinical relevance of SIGLEC-15 as an independent marker of tumor aggressiveness and poor prognosis in gastric adenocarcinoma. Moreover, stratification based on combined SIGLEC-15 and PD-L1 CPS expression revealed that patients co-expressing high levels of both markers experienced the poorest survival outcomes. These findings suggest that the dual assessment of SIGLEC-15 and PD-L1 may enhance prognostic accuracy and support immunotherapeutic decision-making in gastric cancer.
Asunto(s)
Adenocarcinoma , Antígeno B7-H1 , Lectinas , Neoplasias Gástricas , Humanos , Neoplasias Gástricas/patología , Neoplasias Gástricas/metabolismo , Neoplasias Gástricas/mortalidad , Neoplasias Gástricas/genética , Antígeno B7-H1/metabolismo , Antígeno B7-H1/genética , Masculino , Femenino , Persona de Mediana Edad , Pronóstico , Anciano , Biomarcadores de Tumor/metabolismo , Lectinas/metabolismo , Lectinas/genética , Estudios Retrospectivos , Adulto , Anciano de 80 o más Años , Adenocarcinoma/patología , Adenocarcinoma/metabolismo , Inmunoglobulinas , Proteínas de la MembranaRESUMEN
Extracellular matrix (ECM) structures in the brain, including perineuronal nets (PNNs), contribute to synaptic plasticity, neuroprotection, and circuit stability. Wisteria floribunda agglutinin (WFA) is widely used to label PNNs but detects only a subset of ECM glycoconjugates. To better understand ECM composition and organization, we used a panel of plant-derived lectins with distinct glycan-binding specificities to examine the spatial distribution of ECM in the mouse primary somatosensory cortex and hippocampus. Brain sections from adult male C57BL/6N mice (n = 6) were stained using seven biotinylated lectins (Jacalin, PSA, AAL, LCA, WGA, PHA-E, and PHA-L). Confocal imaging and quantitative fluorescence analysis revealed that each lectin exhibited a unique pattern of ECM labeling. Several lectins, such as Jacalin, WGA, and PHA-E, showed strong labeling in superficial cortical layers and select hippocampal subfields, while others displayed more diffuse or subregion-specific distributions. Co-labeling with WFA demonstrated partial overlap with some lectins but also revealed non-overlapping ECM components. Notably, most lectin signals, including those forming perineuronal net-like structures, remained intact after chondroitinase ABC digestion, in contrast to the loss of WFA staining. These results highlight the molecular heterogeneity and anatomical specificity of brain ECM structures, revealing a chondroitin sulfate-independent scaffold not visualized by WFA. These results highlight the molecular heterogeneity and anatomical specificity of brain ECM structures. Using diverse lectins provides complementary information to WFA-based labeling and reveals broader ECM architectures. This approach offers new insights into region- and layer-specific ECM organization, with implications for understanding the differential plasticity, resilience, and vulnerability of brain circuits.
Asunto(s)
Matriz Extracelular , Hipocampo , Lectinas , Corteza Somatosensorial , Animales , Matriz Extracelular/metabolismo , Masculino , Hipocampo/metabolismo , Ratones Endogámicos C57BL , Corteza Somatosensorial/metabolismo , Lectinas de Plantas/metabolismo , Ratones , Lectinas/metabolismo , Condroitina ABC Liasa/metabolismo , Receptores N-AcetilglucosaminaRESUMEN
Omentin-1 (OMNT1) is a metabolically active adipokine implicated in endocrine regulation; however, its role in the anterior pituitary (AP) remains unknown. We hypothesized that OMNT1 modulates the endocrine function of AP cells through gene- and protein-level mechanisms, with effects depending on the animal's metabolic background. To investigate this hypothesis, AP cells isolated from two pig breeds with distinct metabolic profiles, Large White (LW; normal weight) and Meishan (MS; genetically obese), were treated with OMNT1. Transcriptomic and proteomic analyses were performed alongside assessments of hormone expression and secretion. Transcriptomic profiling revealed 13 310 and 13 272 expressed genes in LW and MS pigs, respectively. Differentially expressed gene (DEG) analysis revealed 655 DEGs in LW pigs and 420 in MS pigs. Integrated transcriptomic and proteomic analyses revealed that OMNT1 modulates pathways involved in cellular signaling, cytoskeleton dynamics, responses to stimuli, intracellular protein transport, and post-translational modifications. We further examined the mRNA expression and secretion of tropic hormones (GH, PRL, TSH, ACTH, LH, and FSH) and selected adipokines (adiponectin, leptin, chemerin, apelin, visfatin, resistin, and vaspin), along with their specific receptors. OMNT1 increased LH secretion and decreased FSH levels in a breed-dependent manner. Additionally, in MS pigs, OMNT1 reduced adiponectin and increased leptin secretion. These findings highlight the role of OMNT1 as a key modulator of pituitary endocrine activity, integrating metabolic signals through breed-specific molecular responses at the transcriptional, proteomic, and functional levels.
Asunto(s)
Citocinas , Lectinas , Adenohipófisis , Proteoma , Transcriptoma , Animales , Porcinos , Lectinas/genética , Lectinas/farmacología , Lectinas/metabolismo , Proteómica/métodos , Citocinas/genética , Citocinas/metabolismo , Citocinas/farmacología , Adenohipófisis/metabolismo , Adenohipófisis/citología , Adenohipófisis/efectos de los fármacos , Perfilación de la Expresión Génica , Células CultivadasRESUMEN
Glycan-mediated molecular recognition is crucial for life. Various methodologies, including nuclear magnetic resonance (NMR), help elucidate these interactions across different complexity levels, from macroscopic to atomic resolution. NMR is widely used to study glycan binding to lectins in solution, though these interactions are typically weak (mM to µM scale) under diluted conditions. In nature, multivalent presentations enhance affinity, making interactions more promiscuous.Herein, we describe an NMR methodology, from the ligand perspective, to investigate glycan-lectin interactions in environments mimicking native cellular conditions. For lectins primarily found on cell surfaces, saturation transfer difference (STD)-NMR was used to examine interactions.This NMR methodology provides a proof of concept for studying glycan-lectin interactions in biologically relevant environments. The ability to detect glycan recognition events on the cell surface expands our understanding of these essential interactions and their implications for biomedical applications, including cancer immunotherapy.
Asunto(s)
Lectinas , Resonancia Magnética Nuclear Biomolecular , Polisacáridos , Polisacáridos/metabolismo , Polisacáridos/química , Unión Proteica , Humanos , Lectinas/metabolismo , Lectinas/química , Espectroscopía de Resonancia Magnética/métodos , Ligandos , Resonancia Magnética Nuclear Biomolecular/métodosRESUMEN
Carbohydrate-protein interactions are important in cell-cell communication, signal transduction, cancer, or infection. Chemists have designed glycosylated multivalent systems to mimic these recognition phenomena and produce potent ligands of lectins with therapeutic applications. Dynamic combinatorial chemistry (DCC) provides access to libraries of glycosylated macrocycles equilibrating through reversible covalent bonds. This strategy can be applied to the rapid and efficient identification of multivalent glycoclusters by introducing a protein into the equilibrating library. This strategy allowed the identification of the best ligands for more than one lectin in a single experimental set up by using two simple 1,4-dithiophenol building blocks. Selection of the best binder by each lectin (ConA, LecA, and LecB) was accompanied by the amplification of glyco-dyn[3]arenes and glyco-dyn[4]arenes. These macrocycles could be synthesized, isolated, and displayed nanomolar dissociation constants. Furthermore, while no toxicity could be detected against human cells or bacteria, their anti-adhesive properties against Pseudomonas aeruginosa were confirmed through a virulence assay on human cells. Altogether, extremely simple 1,4-dithiophenol building blocks provided access to a large diversity of glycoconjugates that could be selected by a lectin in a simple experimental set up to identify glycoconjugates with potential anti-infectious applications, thus speeding up the discovery of potential new antibacterial treatments.
Asunto(s)
Antibacterianos , Calixarenos , Glicoconjugados , Fenoles , Pseudomonas aeruginosa , Pseudomonas aeruginosa/efectos de los fármacos , Pseudomonas aeruginosa/química , Glicoconjugados/química , Glicoconjugados/farmacología , Glicoconjugados/síntesis química , Humanos , Técnicas Químicas Combinatorias , Lectinas/química , Lectinas/metabolismo , Calixarenos/química , Antibacterianos/química , Antibacterianos/farmacología , Antibacterianos/síntesis química , Ligandos , Fenoles/química , Adhesión Bacteriana/efectos de los fármacos , Concanavalina A/química , Adhesinas BacterianasRESUMEN
SIGLEC15 is a novel immunosuppressive transmembrane protein promotes cancer development. However, the role of SIGLEC15 in cervical cancer remains poorly understood until now. In this study, we identified that the expression of SIGLEC15 was significantly increased in cervical cancer cells and tissues, and it was closely related to the metastasis of cervical cancer (70.6 %). Several anti-SIGLEC15 mAbs (named 6E3, 2B7, 3G2, 4F6, 4C4, 2F6) were prepared and identified, the titers were from 1:64000 to 1:128000. The results of IFA, IPMA, and IHC revealed that 6E3 (exhibiting a Kaff of 281,200,000 L/mol) had the capability to bind to cervical cancer cells and tissues. Then, the quantum dot nanoparticles probe (QDs-6E3) was synthesized and optimized. Moreover, the QDs based Immunochromatography Assay (QDs-ICA) method for the detection of SIGLEC15 was developed. After several evaluations, SIGLEC15 proved detectable at concentrations as low as 3 ng/mL within 4 min, demonstrating no cross-reactivity. The QDs-ICA exhibited a specificity of 96.7 % and a sensitivity of 96 %. The QDs-ICA for detecting SIGLEC15 is of great significance for the non-invasive screening of patients in clinical trials and the accurate evaluation of the therapeutic effect of this target in cervical cancer.
Asunto(s)
Cromatografía de Afinidad , Lectinas , Proteínas de la Membrana , Neoplasias del Cuello Uterino , Humanos , Femenino , Neoplasias del Cuello Uterino/metabolismo , Neoplasias del Cuello Uterino/patología , Neoplasias del Cuello Uterino/diagnóstico , Lectinas/inmunología , Lectinas/metabolismo , Proteínas de la Membrana/inmunología , Proteínas de la Membrana/metabolismo , Cromatografía de Afinidad/métodos , Anticuerpos Monoclonales/inmunología , Línea Celular Tumoral , InmunoglobulinasRESUMEN
Silurus asotus (Amur catfish) egg lectin (SAL) inhibits cell proliferation and enhances the effects of anticancer drugs by binding to globotriaosylceramide (Gb3) on the cell surface. Gb3 expression is typically increased in seminomas. However, its association with SAL and the underlying mechanisms remain unclear. Here, we investigated the effects of SAL on morphology, migratory ability, and integrin expression in JKT-1 cells using chromatography and mass spectrometry. Gb3 was expressed in JKT-1, an established seminoma cell line. SAL did not alter JKT-1 proliferation but increased propidium iodide uptake. Furthermore, SAL induced morphological changes and increased the expression of integrin α2 in JKT-1, but not in HeLa cells. Gb3 expression was detected in JKT-1 and HeLa cells, with high- and low-mobility bands observed. However, the low-mobility bands were more abundant in JKT-1 than in HeLa cells. The main forms of Gb3 in JKT-1 cells were high-mobility d18:1/24:0 and d18:1/24:1 and low-mobility hydroxylated Gb3. Fatty acid 2-hydroxylase was involved in the acyl chain hydroxylation of low-mobility Gb3 in JKT-1 cells and showed 5-fold higher expression in JKT-1 cells than in HeLa cells. Our findings suggest that the antitumor effects of SAL vary according to the specific Gb3 molecular species expressed in cancer cells.
Asunto(s)
Bagres , Lectinas , Trihexosilceramidas , Animales , Humanos , Células HeLa , Lectinas/farmacología , Lectinas/metabolismo , Bagres/metabolismo , Trihexosilceramidas/metabolismo , Proliferación Celular/efectos de los fármacos , Línea Celular Tumoral , Movimiento Celular/efectos de los fármacosRESUMEN
This study aimed to compare subcutaneous and visceral adipocyte geometries, clinical, metabolic, hormonal, and gene expression parameters, between participants with insulin resistance (IR; n = 6) and without IR (n = 28). Additionally, correlations between these factors and adipocyte geometries were analyzed in 27 participants, including 14 with obesity and 13 without obesity, due to limited tissue availability. In a cross-sectional study at a tertiary hospital, 34 female patients undergoing intra-abdominal surgery were recruited. Participants with IR had higher serum leptin and larger visceral adipocytes but lower serum omentin and adiponectin/leptin ratio, despite comparable gene expression. Visceral adipocyte geometries showed positive correlations with IR parameters (glucose and HOMA-IR) only in individuals without obesity, but exhibited negative correlations with QUICKI, serum adiponectin, adiponectin/leptin ratio, omentin, and/or visfatin in both groups, with generally stronger correlations observed in individuals without obesity. Importantly, enlarged adipocytes were associated with lower subcutaneous and visceral adiponectin, omentin, and visfatin mRNA expression in individuals with obesity, and with higher visceral LEP mRNA expression in individuals without obesity. Multiple regression analysis identified the serum adiponectin/leptin ratio as an independent predictor of adipocyte geometries in both participants with and without obesity, and body weight in individuals without obesity. In conclusion, enlarged visceral adipocytes were more strongly associated with metabolic dysregulation in individuals without obesity, while individuals with obesity exhibited mostly weaker or no correlations, possibly due to reduced capacity for further adipocyte expansion and pre-existing metabolic impairments. The serum adiponectin/leptin ratio is a sensitive biomarker reflecting adipocyte function and morphology.
Asunto(s)
Adipocitos , Grasa Intraabdominal , Obesidad , Grasa Subcutánea , Humanos , Femenino , Obesidad/metabolismo , Obesidad/patología , Obesidad/sangre , Adipocitos/metabolismo , Adipocitos/patología , Grasa Intraabdominal/metabolismo , Grasa Intraabdominal/patología , Persona de Mediana Edad , Adulto , Resistencia a la Insulina , Estudios Transversales , Adiponectina/sangre , Grasa Subcutánea/metabolismo , Grasa Subcutánea/patología , Leptina/sangre , Citocinas/sangre , Citocinas/genética , Citocinas/metabolismo , Nicotinamida Fosforribosiltransferasa/genética , Nicotinamida Fosforribosiltransferasa/sangre , Nicotinamida Fosforribosiltransferasa/metabolismo , Proteínas Ligadas a GPI/sangre , Proteínas Ligadas a GPI/metabolismo , Proteínas Ligadas a GPI/genética , Lectinas/sangre , Lectinas/genética , Lectinas/metabolismoRESUMEN
The Spike glycoprotein of SARS-CoV-2 is the major target for vaccines and therapeutics. Spike glycosylation is critical for ACE2 binding and subsequent viral fusion and entry. Here, we studied lectins for their ability to bind to SARS-CoV-2 Spike glycoprotein and SARS-CoV-2 virions by employing an array of 95 lectins, for 68 of which we predicted glycan-binding specificities using publically available glycan array data and MotifFinder software. We identified lectins with diverse glycan binding specificities that bound with high intensities to recombinant Spike and cultured SARS-CoV-2 virus - AAL, ABL, ACL, AMA, ASA, BANLEC, BC2L-A, RCA 120, CALSEPA, GAL3, GS-II, PALa, CA, HHA, PHA-L, PA-IIL, MNA-M, STL, LSL-N, GRFT, PSA, RS-FUC, PHA-E, CPA, LENTIL, RCA 60, GNA, ORYSATA, LcH A, PHA-P, PTL-2, MAA, Con A, TL, NPA, and SBA. Analyzing the glycan-binding specificities of these lectins, we predict that the Spike glycoprotein is modified with high mannose/hybrid N-glycans with terminal mannose residues, α1-6 core fucosylated N-glycans with terminal GlcNAc residues, and complex glycans with Lewis A, Lewis B, Lewis X, Lewis Y, and Blood group H structures on type-1 or type-2 extension sequences. The SARS-CoV-2-specific lectins identified in our study may be assessed for their antiviral potential in future studies.
Asunto(s)
Lectinas , SARS-CoV-2 , Glicoproteína de la Espiga del Coronavirus , SARS-CoV-2/metabolismo , Glicoproteína de la Espiga del Coronavirus/metabolismo , Glicoproteína de la Espiga del Coronavirus/química , Lectinas/metabolismo , Lectinas/química , Humanos , Unión Proteica , COVID-19/virología , COVID-19/metabolismo , Polisacáridos/metabolismo , Glicosilación , Análisis por Matrices de ProteínasRESUMEN
Protein-carbohydrate interactions are utilized by the immune system for several defense activities. Collectins and ficolins have a key role in maintaining the balance of surfactants in the lungs and supporting the immune system in the lungs, respectively. This chapter provides an extensive description of two soluble pattern recognition receptors (PRRs), collectins and ficolins. It covers their structure, specificity for binding to certain ligands, biological functions along with initiating immune response by various mechanisms, their genetic characteristics, associations with various diseases, and their potential for use in therapy and diagnosis. The potential of collectins as diagnostic markers and therapeutic agents in diverse malignancies for their antitumorigenic effects has also been investigated. These findings have greatly enhanced our understanding of the innate immune system's previously overlooked specificity and effectiveness.
Asunto(s)
Colectinas , Lectinas , Humanos , Lectinas/genética , Lectinas/metabolismo , Lectinas/química , Lectinas/inmunología , Ficolinas , Animales , Colectinas/genética , Colectinas/metabolismo , Colectinas/química , Colectinas/inmunología , Inmunidad Innata , Receptores de Reconocimiento de Patrones/inmunología , Receptores de Reconocimiento de Patrones/metabolismo , Neoplasias/inmunología , Neoplasias/metabolismoRESUMEN
Glycans on the surface of all immune cells are the product of diverse post-translational modifications (glycosylation) that affect almost all proteins and possess enormous structural heterogeneity. Their bioinformational content is decoded by glycan-binding proteins (lectins, GBPs), such as C-type lectins, including selectins, galectins, and Siglecs. Glycans located on the surface of immune cells are involved in many immunological processes through interactions with GBPs. Lectins recognize changes in the glycan epitopes; distinguish among host (self), microbial (non-self), and tumor (modified self) antigens; and consequently regulate immune responses. Understanding GBP-glycan interactions accelerates the development of glycan-targeted therapeutics in severe diseases, including inflammatory and autoimmune diseases and cancer. This review will discuss N- and O-glycosylations and glycosyltransferases involved in the biosynthesis of carbohydrate epitopes and address how interactions between glycan epitopes and GBPs are crucial in immune responses. The pivotal role of the glycan antigen tetrasaccharide sialyl Lewis x in mediating immune and tumor cell trafficking into the extravascular site will be discussed. Next, the role of glycans in modulating bacterial, fungal, viral, and parasitic infections and cancer will be surveyed. Finally, the role of glycosylation in antibodies and carbohydrate vaccines will be analyzed.
Asunto(s)
Inmunidad , Polisacáridos , Humanos , Polisacáridos/inmunología , Polisacáridos/metabolismo , Polisacáridos/química , Glicosilación , Epítopos/inmunología , Animales , Neoplasias/inmunología , Lectinas/metabolismo , Lectinas/inmunología , Procesamiento Proteico-PostraduccionalRESUMEN
Extracellular vesicles (EVs) exhibit extensive glycosylation modifications, which are promising biomarkers for gastric cancer (GC). However, EV glycomics and the potential application of EV glycosylation patterns in liquid biopsy remain largely unexplored. This study aims to elucidate the heterogeneity of EV glycosylation in the initiation and progression of GC and to identify specific EV glycosylation markers for clinical assessment. We developed a novel platform for analyzing EV glycosylation patterns via a lectin microarray for EV glycosyl-lectin affinity assessment and antibody-based EV subgroup annotation. The lectin-affinity glycosylation pattern (LAGP) of plasma EVs was profiled across 84 plasma samples, encompassing cases from different stages of GC, benign gastric diseases (BD), and non-disease control (NC). We uncovered heterogeneous LAGPs in different patient groups, identified group-specific LAGPs, and employed them in modeling with the assistance of machine learning algorithms. The linear discriminant analysis (LDA) distinguished advanced GC, early GC, BD, and NC samples with 100 % accuracy. A LAGP-based nomogram was established to predict survival outcomes within 200, 300, and 500 days, achieving area under the ROC curve (AUC) of 0.793, 0.914, and 0.988, respectively. We also tested LAGP for immunotherapeutic response prediction, obtaining AUC values of 0.866-1.000 with various supervised machine-learning algorithms. In conclusion, we represented heterogeneous LAGPs during the occurrence and progression of GC and developed an all-in-one clinical assessment tool for screening cancer patients, monitoring survival outcomes, and predicting immunotherapeutic responses.