With the size of the biopharmaceutical market exponentially increasing,there is an aligned growth in the importance of data-rich analyses,not only to assess drug product safety but also to assist drug development driv...With the size of the biopharmaceutical market exponentially increasing,there is an aligned growth in the importance of data-rich analyses,not only to assess drug product safety but also to assist drug development driven by the deeper understanding of structure/function relationships.In monoclonal antibodies,many functions are regulated by N-glycans present in the constant region of the heavy chains and their mechanisms of action are not completely known.The importance of their function focuses analytical research efforts on the development of robust,accurate and fast methods to support drug development and quality control.Released N-glycan analysis is considered as the gold standard for glycosylation characterisation;however,it is not the only method for quantitative analysis of glycoform heterogeneity.In this study,ten different analytical workflows for N-glycan analysis were compared using four monoclonal antibodies.While observing good comparability between the quantitative results generated,it was possible to appreciate the advantages and disadvantages of each technique and to summarise all the observations to guide the choice of the most appropriate analytical workflow according to application and the desired depth of data generated.展开更多
Malignant tumors are complex structures composed of cancer cells and tumor microenvironmental cells.In this complex structure,cells cross-talk and interact,thus jointly promoting cancer development and metastasis.Rece...Malignant tumors are complex structures composed of cancer cells and tumor microenvironmental cells.In this complex structure,cells cross-talk and interact,thus jointly promoting cancer development and metastasis.Recently,immunoregulatory molecule-based cancer immunotherapy has greatly improved treatment efficacy for solid cancers,thus enabling some patients to achieve persistent responses or cure.However,owing to the development of drug-resistance and the low response rate,immunotherapy against the available targets PD-1/PD-L1 or CTLA-4 has limited benefits.Although combination therapies have been proposed to enhance the response rate,severe adverse effects are observed.Thus,alternative immune checkpoints must be identified.The SIGLECs are a family of immunoregulatory receptors(known as glyco-immune checkpoints)discovered in recent years.This review systematically describes the molecular characteristics of the SIGLECs,and discusses recent progress in areas including synthetic ligands,monoclonal antibody inhibitors,and Chimeric antigen receptor T(CAR-T)cells,with a focus on available strategies for blocking the sialylated glycan-SIGLEC axis.Targeting glyco-immune checkpoints can expand the scope of immune checkpoints and provide multiple options for new drug development.展开更多
BACKGROUND Hepatitis B virus (HBV) infection is the primary cause of hepatitis with chronic HBV infection,which may develop into liver fibrosis,cirrhosis and hepatocellular carcinoma.Detection of early-stage fibrosis ...BACKGROUND Hepatitis B virus (HBV) infection is the primary cause of hepatitis with chronic HBV infection,which may develop into liver fibrosis,cirrhosis and hepatocellular carcinoma.Detection of early-stage fibrosis related to HBV infection is of great clinical significance to block the progression of liver lesion.Direct liver biopsy is regarded as the gold standard to detect and assess fibrosis;however,this method is invasive and prone to clinical sampling error.In order to address these issues,we attempted to find more convenient and effective serum markers for detecting HBV-induced early-stage liver fibrosis.AIM To investigate serum N-glycan profiling related to HBV-induced liver fibrosis and verify multiparameter diagnostic models related to serum N-glycan changes.METHODS N-glycan profiles from the sera of 432 HBV-infected patients with liver fibrosis were analyzed.Significant changed N-glycan levels (peaks)(P <0.05) in differentfibrosis stages were selected in the modeling group,and multiparameter diagnostic models were established based on changed N-glycan levels by logistic regression analysis.The receiver operating characteristic (ROC) curve analysis was performed to evaluate diagnostic efficacy of N-glycans models.These models were then compared with the aspartate aminotransferase to platelet ratio index (APRI),fibrosis index based on the four factors (FIB-4),glutamyltranspeptidase platelet albumin index (S index),GlycoCirrho-test,and GlycoFibro-test.Furthermore,we combined multiparameter diagnostic models with alanine aminotransferase (ALT) and platelet (PLT) tests and compared their diagnostic power.In addition,the diagnostic accuracy of N-glycan models was also verified in the validation group of patients.RESULTS Multiparameter diagnostic models constructed based on N-glycan peak 1,3,4and 8 could distinguish between different stages of liver fibrosis.The area under ROC curves (AUROCs) of Model A and Model B were 0.890 and 0.752,respectively differentiating fibrosis F0-F1 from F2-F4,and F0-F2 from F3-F4,and surpassing other serum panels.However,AUROC (0.747) in Model C used for the diagnosis of F4 from F0-F3 was lower than AUROC (0.795) in FIB-4.In combination with ALT and PLT,the multiparameter models showed better diagnostic power (AUROC=0.912,0.829,0.885,respectively) when compared with other models.In the validation group,the AUROCs of the three combined models (0.929,0.858,and 0.867,respectively) were still satisfactory.We also applied the combined models to distinguish adjacent fibrosis stages of 432patients (F0-F1/F2/F3/F4),and the AUROCs were 0.917,0.720 and 0.785.CONCLUSION Multiparameter models based on serum N-glycans are effective supplementary markers to distinguish between adjacent fibrosis stages of patients caused by HBV,especially in combination with ALT and PLT.展开更多
Bacterial surface glycans perform a diverse and important set of biological roles,and have been widely used in the treatment of bacterial infectious diseases.The majority of bacterial surface glycans are decorated wit...Bacterial surface glycans perform a diverse and important set of biological roles,and have been widely used in the treatment of bacterial infectious diseases.The majority of bacterial surface glycans are decorated with diverse rare functional groups,including amido,acetamidino,carboxamido and pyruvate groups.These functional groups are thought to be important constituents for the biological activities of glycans.Chemical synthesis of glycans bearing these functional groups or their variants is essential for the investigation of structure-activity relationships by a medicinal chemistry approach.To date,a broad choice of synthetic methods is available for targeting the different rare functional groups in bacterial surface glycans.This article reviews the structures of naturally occurring rare functional groups in bacterial surface glycans,and the chemical methods used for installation of these groups.展开更多
Rotavirus(RV)causes acute gastroenteritis in infants and children worldwide.Recent studies showed that glycans such as histo-blood group antigens(HBGAs)function as cell attachment factors affecting RV host susceptibil...Rotavirus(RV)causes acute gastroenteritis in infants and children worldwide.Recent studies showed that glycans such as histo-blood group antigens(HBGAs)function as cell attachment factors affecting RV host susceptibility and prevalence.P[8]is the predominant RV genotype in humans,but the structural basis of how P[8]RVs interact with glycan ligands remains elusive.In this study,we characterized the interactions between P[8]VP8~*s and glycans which showed that VP8~*,the RV glycan binding domain,recognized both mucin core 2 and H type 1 antigens according to the ELISA-based oligosaccharide binding assays.Importantly,we determined the structural basis of P[8]RV-glycans interaction from the crystal structures of a Rotateq P[8]VP8~*in complex with core 2 and H type 1 glycans at 1.82.3?,respectively,revealing a common binding pocket and similar binding mode.Structural and sequence analysis demonstrated that the glycan binding site is conserved among RVs in the P[Ⅱ]genogroup,while genotype-specific amino acid variations determined different glycan binding preference.Our data elucidated the detailed structural basis of the interactions between human P[8]RVs and different host glycan factors,shedding light on RV infection,epidemiology,and development of anti-viral agents.展开更多
Lassa virus(LASV)belongs to the Mammarenavirus genus(family Arenaviridae)and causes severe hemorrhagic fever in humans.The glycoprotein complex(GPC)contains eleven N-linked glycans that play essential roles in GPC fun...Lassa virus(LASV)belongs to the Mammarenavirus genus(family Arenaviridae)and causes severe hemorrhagic fever in humans.The glycoprotein complex(GPC)contains eleven N-linked glycans that play essential roles in GPC functionalities such as cleavage,transport,receptor recognition,epitope shielding,and immune response.We used three mutagenesis strategies(asparagine to glutamine,asparagine to alanine,and serine/tyrosine to alanine mutants)to abolish individual glycan chain on GPC and found that all the three strategies led to cleavage inefficiency on the 2nd(N89),5th(N119),or 8th(N365)glycosylation motif.To evaluate N to Q mutagenesis for further research,it was found that deletion of the 2nd(N89Q)or 8th(N365Q)glycan completely inhibited the transduction efficiency of pseudotyped particles.We further investigated the role of individual glycan on GPC-mediated immune response by DNA immunization of mice.Deletion of the individual 1st(N79Q),3rd(N99Q),5th(N119Q),or 6th(N167Q)glycan significantly enhanced the proportion of effector CD4+cells,whereas deletion of the 1st(N79Q),2nd(N89Q),3rd(N99Q),4th(N109Q),5th(N119Q),6th(N167Q),or 9th(N373Q)glycan enhanced the proportion of CD8+effector T cells.Deletion of specific glycan improves the Th1-type immune response,and abolishment of glycan on GPC generally increases the antibody titer to the glycan-deficient GPC.However,the antibodies from either the mutant or WT GPC-immunized mice show little neutralization effect on wild-type LASV.The glycan residues on GPC provide an immune shield for the virus,and thus represent a target for the design and development of a vaccine.展开更多
Sialylation is one of important glycosylation in human beings and plays an important role in cancer development. α2,3-Linked and α2,6-linked sialic acids are normally observed on the end of N-glycans and have differ...Sialylation is one of important glycosylation in human beings and plays an important role in cancer development. α2,3-Linked and α2,6-linked sialic acids are normally observed on the end of N-glycans and have different functions. Derivatization on sialic acid was designed to detect the different linkages by MALDI-TOF MS. In this study, a two-step derivatization by dimethylamine and ammonium hydroxide was improved to modify the sialic acid and made it easier to detect the different linkages of sialic acids on MALDI-TOF MS. Using this derivatization method, specific sialic acids linkages on N-glycans of protein samples such as fetuin and lactoferrin were detected. For complex cell samples, increased a2,3-linked and a2,6-linked sialic acids on bi-antennary and tri-antennary N-glycans were observed in A549 cells induced by hypoxia environment. Taken together, our two-step derivatization of sialic acids offers a simple and accurate way to detect specific linkages on N-glycans with MALDI-TOF mass spectrometer.展开更多
To explore the effects of traditional herbal medicine Ganoderrna tsugae（G, tsugae） on immunomodulatory and antitumor activities, the crude polysaccharides of G. tsugae were purified by filtration, diethylaminoethyl...To explore the effects of traditional herbal medicine Ganoderrna tsugae（G, tsugae） on immunomodulatory and antitumor activities, the crude polysaccharides of G. tsugae were purified by filtration, diethylaminoethyl（DEAE） sepharose-fast flow chromatography and sephadex G-100 size-exclusion chromatography. Two main fractions, pro- tein-containing glyeans CSSLP-1 and CSSLP-2, were obtained via the gradient elution. The protein content, molecu- lar weight, and monosaccharide composition of the two fractions were analyzed. Furthermore, the influence of the protein-containing glycans from G. tsugae on the activation of human acute monocytic leukemia cell line（THP-l） and their antitumor activities to the human hepatocellular liver carcinoma cell（HepG-2） in vitro were evaluated. The re- sults indicate that CSSLP-1 and CSSLP-2 could increase the pinocytie activity of THP-1 cells and induce THP-1 cells to produce the eytokines of TNFa and IL-2, significantly. CSSLP-1 and CSSLP-2 also played an inhibiting effect on the cancer cell（HepG-2）. Moreover, the anti-proliferation activity of CSSLP-1 and CSSLP-2 increased with the par- ticipation of TNFa and IL-2 or other antitumor factors induced from THP-1 cells by G. tsugae protein-containing glycan fractions.展开更多
Spermatogenesis, maturation, capacitation and fertilization are precisely regulated by glycosylation. However, the relationship between altered glycosylation patterns and the onset and development of reproductive diso...Spermatogenesis, maturation, capacitation and fertilization are precisely regulated by glycosylation. However, the relationship between altered glycosylation patterns and the onset and development of reproductive disorders is unclear, mainly limited by the lack of in situ imaging techniques for spermatozoa glycosylation. We developed an efficient and highly specific spermatozoa glycan imaging technique based on the robust chemoselective labeling of sialic acid(Sia) and N-acetyl-D-galactosamine(Gal/GalNAc). We further proposed a “tandem glycan chemoselective labeling” strategy to achieve simultaneous imaging of two types of glycans on spermatozoa. We applied the developed method to the spermatozoa from oligozoospermic patients and diabetic mice and found that these spermatozoa showed higher levels of Sia and Gal/Gal NAc expression than the normal groups. Moreover, spermatozoa from diabetic mice showed a severe decrease in number, viability, and forward motility, suggesting that in vivo glucose metabolism disorders may lead to an elevated level of spermatozoa glycosylation and have a correlation with the development of oligoasthenotspermia. Our work provides a research tool to reveal the relationship between glycosylation modification and spermatozoa quality, and a promising clue for the development of glycan-based reproductive markers.展开更多
Background Optimal gut health is important to maximize growth performance and feed efficiency in broiler chickens.A total of 1,365 one-day-old male Ross 308 broiler chickens were randomly divided into 5 treatments gro...Background Optimal gut health is important to maximize growth performance and feed efficiency in broiler chickens.A total of 1,365 one-day-old male Ross 308 broiler chickens were randomly divided into 5 treatments groups with 21 replicates,13 birds per replicate.The present research investigated effects of microbial muramidase or a precision glycan alone or in combination on growth performance,apparent total tract digestibility,total blood carotenoid content,intestinal villus length,meat quality and gut microbiota in broiler chickens.Treatments included:NC:negative control(basal diet group);PC:positive control(basal diet+0.02%probiotics);MR:basal diet+0.035%microbial muramidase;PG:basal diet+0.1%precision glycan;and MRPG:basal diet+0.025%MR+0.1%PG,respectively.Results MRPG group increased the body weight gain and feed intake(P<0.05)compared with NC group.Moreover,it significantly increased total serum carotenoid(P<0.05)and MRPG altered the microbial diversity in ileum contents.The MRPG treatment group increased the abundance of the phylum Firmicutes,and family Lachnospiraceae,Ruminococcaceae,Oscillospiraceae,Lactobacillaceae,Peptostreptococcaceae and decreased the abundance of the phylum Campilobacterota,Bacteroidota and family Bacteroidaceae.Compared with the NC group,the chickens fed MRPG showed significantly increased in duodenum villus length at end the trial.Conclusion In this study,overall results showed that the synergetic effects of MR and PG showed enhancing growth performance,total serum carotenoid level and altering gut microbiota composition of broilers.The current research indicates that co-supplementation of MR and PG in broiler diets enhances intestinal health,consequently leading to an increased broiler production.展开更多
Native and methyl-esterified sialylated glycans were analyzed with 2,4,6-trihydroxyacetophenone（THAP）and 2,5-dihydroxybenzoic acid（DHB）as matrix by a matrix-assisted laser desorption/ionization time-of-flight mass...Native and methyl-esterified sialylated glycans were analyzed with 2,4,6-trihydroxyacetophenone（THAP）and 2,5-dihydroxybenzoic acid（DHB）as matrix by a matrix-assisted laser desorption/ionization time-of-flight mass spectrometer（MALDI-TOF MS）.High quality negative-ion spectra of commercial sialylated glycan were obtained with THAP as matrix.Detection limit of the glycan was less than 0.1 pmol.After methyl esterification of sialic acid（SA）residue,sialylated glycans were detected sensitively in the positive-ion mode using DHB as matrix.Neutral and sialylated glycans from the mixture of asialofetuin and fetuin were methylesterified and simultaneously recognized in one manipulation.Methyl esterification of SA residue offers a convenient and sensitive way to identify the structure of N-linked glycans for glycan profiling.展开更多
As the two most principal active substances in the corn silk,polysaccharides and flavonoids,the mechanism of interaction between them has been a topic of intense research.This study provides an in-depth investigation ...As the two most principal active substances in the corn silk,polysaccharides and flavonoids,the mechanism of interaction between them has been a topic of intense research.This study provides an in-depth investigation of the interaction mechanism between corn silk glycans and luteoloside(LUT)and the synergistic role that result from this interaction.The interaction mechanism was evaluated by isothermal titration calorimetry(ITC)and circular dichroism(CD),and the synergistic role was evaluated by the expression of glucose transporters(GLUT-1),insulin secretion and surface plasmon resonance(SPR).CD and ITC results indicated that the interaction between CSGs and LUT mainly driven by the Cotton effects,enthalpy and entropy-driven.This interaction precipitated the formation of complexes(CSGs/LUT complexes)between corn silk glycans(CSGs)with four different molecular weights and luteoloside(LUT).Furthermore,the CSGs and LUT play a synergistic role in glucose regulation through GLUT-1 expression and insulin secretion experiments,compared to single luteoloside group.展开更多
The glycosylation of proteins is responsible for their structural and functional roles in many cellular activities.This work describes a strategy that combines an efficient release, labeling and liquid chromatography...The glycosylation of proteins is responsible for their structural and functional roles in many cellular activities.This work describes a strategy that combines an efficient release, labeling and liquid chromatography–mass spectral analysis with the use of a comprehensive database to analyze N-glycans. The analytical method described relies on a recently commercialized kit in which quick deglycosylation is followed by rapid labeling and cleanup of labeled glycans. This greatly improves the separation, mass spectrometry（MS） analysis and fluorescence detection of N-glycans. A hypothetical database, constructed using Glyc Resoft, provides all compositional possibilities of N-glycans based on the common sugar residues found in N-glycans. In the initial version this database contains ＆gt; 8,700 N-glycans, and is compatible with MS instrument software and expandable. N-glycans from four different well-studied glycoproteins were analyzed by this strategy. The results provided much more accurate and comprehensive data than that had been previously reported. This strategy was then used to analyze the N-glycans present on the membrane glycoproteins of gastric carcinoma cells with different degrees of differentiation. Accurate and comprehensive N-glycan data from those cells was obtained efficiently and their differences were compared corresponding to their differentiation states. Thus, the novel strategy developed greatly improves accuracy, efficiency and comprehensiveness of N-glycan analysis.展开更多
Impact statement.Spindle-shaped viruses exclusively infect archaea.Fuselloviruses represent a large group of spindle-shaped viruses and infect hyperthermophilic archaea of the order Sulfolobales.Although the first fus...Impact statement.Spindle-shaped viruses exclusively infect archaea.Fuselloviruses represent a large group of spindle-shaped viruses and infect hyperthermophilic archaea of the order Sulfolobales.Although the first fusellovirus was identified nearly 40 years ago,the mechanism of host infection by these viruses remains poorly understood.Here,we show that SSV19,a fusellovirus isolated from a hot spring in the Philippines,is capable of hydrolyzing the host cell surface glycan identified as a heptasaccharide chain of QuiS_(1)Hex_(4)HexNAc_(2).Our findings provide significant insights into the molecular strategy of host recognition and,possibly,entry by an archaeal virus.展开更多
Coronaviruses are enveloped RNA viruses distinguished by their crown-like surface spikes. These viruses cause various diseases in humans and animals, including respiratory, gastrointestinal, and neurological disorders...Coronaviruses are enveloped RNA viruses distinguished by their crown-like surface spikes. These viruses cause various diseases in humans and animals, including respiratory, gastrointestinal, and neurological disorders. The health risks posed by coronaviruses are substantial, as demonstrated by severe acute respiratory syndrome-coronavirus (SARS-CoV) and SARS-CoV-2. SARS-CoV triggered a global outbreak of SARS in 2003, and SARS-CoV-2 has been responsible for the ongoing coronavirus disease 2019 (COVID-19) pandemic since the end of 2019. Furthermore, four human coronaviruses (HCoVs), HCoV-229E, HCoV-OC43, HCoV-NL63, and HCoV-HKU1, have been implicated in 15% to 30% cases of the common cold.[1] Given their prevalence and potential to cause various diseases, understanding the mechanisms underlying the invasion into cells of the coronavirus is essential for public health. By binding to specific cell receptors, the spike protein of coronaviruses undergoes conformational changes that promote membrane fusion and entry of the virus into host cells. Although significant progress has been made in elucidating the mechanisms of coronavirus entry, the detailed molecular interactions between viral proteins and host receptors remain incompletely understood.展开更多
Fluorescent DNA probes commonly used for in situ imaging of cell surface glycans are“always-on”probes(AOPs),which produce high background noise and lack spatial specificity.Molecular dynamics simulations indicate th...Fluorescent DNA probes commonly used for in situ imaging of cell surface glycans are“always-on”probes(AOPs),which produce high background noise and lack spatial specificity.Molecular dynamics simulations indicate that single-stranded DNA(ssDNA)can stably associate with the cell membrane,contributing to persistent background signals in AOPs.To overcome this challenge,a stepwise activated probe(SAP)is developed for in situ imaging of cell surface glycans.In SAP,the cell surface glycan is labeled by rolling circle amplification product through metabolic glycan labeling strategies.The product hybridizes a fluorophore and quencher-labeled ssDNA reporter to form a double strand producing primary enhancement of fluorescence signal.Nicking enzyme(Nb.BbvCI)cleaves the double strand and releases the quencher,further enhancing the signal.Nonspecific absorption of the reporter on the cellular membrane does not increase the fluorophore-quencher distance in SAP or trigger Nb.BbvCI cleavage.As a result,only the glycan sites are illuminated.SAP not only exhibits high imaging specificity but also greatly simplifies the imaging procedure by reducing the washing steps.SAP offers a considerable improvement in detection specificity and sensitivity by employing a two-step activation and amplification process,making it a powerful method for in situ fluorescence imaging.展开更多
Among the leading methods for triggering therapeutic anti-cancer immunity is the inhibition of immune checkpoint pathways.N-glycosylation is found to be essential for the function of various immune checkpoint proteins...Among the leading methods for triggering therapeutic anti-cancer immunity is the inhibition of immune checkpoint pathways.N-glycosylation is found to be essential for the function of various immune checkpoint proteins,playing a critical role in their stability and interaction with immune cells.Removing the N-glycans of these proteins seems to be an alternative therapy,but there is a lack of a de-N-glycosylation technique for target protein specificity,which limits its clinical application.Here,we developed a novel technique for specifically removing N-glycans from a target protein on the cell surface,named deglycosylation targeting chimera(DGlyTAC),which employs a fusing protein consisting of Peptide-N-glycosidase F(PNGF)and target-specific nanobody/affibody(Nb/Af).The DGlyTAC technique was developed to target a range of glycosylated surface proteins,especially these immune checkpoints—CD24,CD47,and PD-L1,which minimally affected the overall N-glycosylation landscape and the N-glycosylation of other representative membrane proteins,ensuring high specificity and minimal off-target effects.Importantly,DGlyTAC technique was successfully applied to lead inactivation of these immune checkpoints,especially PD-L1,and showed more potential in cancer immunotherapy than inhibitors.Finally,PD-L1 targeted DGlyTAC showed therapeutic effects on several tumors in vivo,even better than PD-L1 antibody.Overall,we created a novel target-specific N-glysocylation erasing technique that establishes a modular strategy for directing membrane proteins inactivation,with broad implications on tumor immune therapeutics.展开更多
The immune system is coordinated by an intricate network of stimulatory and inhibitory circuits that regulate host responses against endogenous and exogenous insults.Disruption of these safeguard and homeostatic mecha...The immune system is coordinated by an intricate network of stimulatory and inhibitory circuits that regulate host responses against endogenous and exogenous insults.Disruption of these safeguard and homeostatic mechanisms can lead to unpredictable inflammatory and autoimmune responses,whereas deficiency of immune stimulatory pathways may orchestrate immunosuppressive programs that contribute to perpetuate chronic infections,but also influence cancer development and progression.Glycans have emerged as essential components of homeostatic circuits,acting as fine-tuners of immunological responses and potential molecular targets for manipulation of immune tolerance and activation in a wide range of pathologic settings.Cell surface glycans,present in cells,tissues and the extracellular matrix,have been proposed to serve as“self-associated molecular patterns”that store structurally relevant biological data.The responsibility of deciphering this information relies on different families of glycan-binding proteins(including galectins,siglecs and C-type lectins)which,upon recognition of specific carbohydrate structures,can recalibrate the magnitude,nature and fate of immune responses.This process is tightly regulated by the diversity of glycan structures and the establishment of multivalent interactions on cell surface receptors and the extracellular matrix.Here we review the spatiotemporal regulation of selected glycan-modifying processes including mannosylation,complex N-glycan branching,core 2 O-glycan elongation,LacNAc extension,as well as terminal sialylation and fucosylation.Moreover,we illustrate examples that highlight the contribution of these processes to the control of immune responses and their integration with canonical tolerogenic pathways.Finally,we discuss the power of glycans and glycan-binding proteins as a source of immunomodulatory signals that could be leveraged for the treatment of autoimmune inflammation and chronic infection.展开更多
The pandemic of the severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)has caused a high number of deaths in the world.To combat it,it is necessary to develop a better understanding of how the virus infects ho...The pandemic of the severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)has caused a high number of deaths in the world.To combat it,it is necessary to develop a better understanding of how the virus infects host cells.Infection normally starts with the attachment of the virus to cell-surface glycans like heparan sulfate(HS)and sialic acid-containing glycolipids/glycoproteins.In this study,we examined and compared the binding of the subunits and spike(S)proteins of SARS-CoV-2,SARS-Co V,and Middle East respiratory disease(MERS)-Co V to these glycans.Our results revealed that the S proteins and subunits can bind to HS in a sulfation-dependent manner and no binding with sialic acid residues was detected.Overall,this work suggests that HS binding may be a general mechanism for the attachment of these coronaviruses to host cells,and supports the potential importance of HS in infection and in the development of antiviral agents against these viruses.展开更多
文摘With the size of the biopharmaceutical market exponentially increasing,there is an aligned growth in the importance of data-rich analyses,not only to assess drug product safety but also to assist drug development driven by the deeper understanding of structure/function relationships.In monoclonal antibodies,many functions are regulated by N-glycans present in the constant region of the heavy chains and their mechanisms of action are not completely known.The importance of their function focuses analytical research efforts on the development of robust,accurate and fast methods to support drug development and quality control.Released N-glycan analysis is considered as the gold standard for glycosylation characterisation;however,it is not the only method for quantitative analysis of glycoform heterogeneity.In this study,ten different analytical workflows for N-glycan analysis were compared using four monoclonal antibodies.While observing good comparability between the quantitative results generated,it was possible to appreciate the advantages and disadvantages of each technique and to summarise all the observations to guide the choice of the most appropriate analytical workflow according to application and the desired depth of data generated.
基金supported by the Shanghai Science and Technology Committee (Grant Nos. 20DZ2201900 to Y.Y. and 23ZR1432500 to W.P.)National Natural Science Foundation of China (Grant Nos. 82072602 to Y.Y.+4 种基金91853121, 21977066, and 22177069 to W.P.)Innovation Foundation of Translational Medicine of Shanghai Jiao Tong University School of Medicine(Grant No. TM202001 to Y.Y.)Collaborative Innovation Center for Clinical and Translational Science by Chinese Ministry of Education&Shanghai (Grant No. CCTS-2022202 to Y.Y.)Shanghai Pilot Program for Basic Research-Shanghai Jiao Tong University (Grant No. 21TQ1400210 to W.P.)Medical-Engineering Interdisciplinary Research Foundation of Shanghai Jiao Tong University (Grant No. YG2022ZD001 to W.P.)
文摘Malignant tumors are complex structures composed of cancer cells and tumor microenvironmental cells.In this complex structure,cells cross-talk and interact,thus jointly promoting cancer development and metastasis.Recently,immunoregulatory molecule-based cancer immunotherapy has greatly improved treatment efficacy for solid cancers,thus enabling some patients to achieve persistent responses or cure.However,owing to the development of drug-resistance and the low response rate,immunotherapy against the available targets PD-1/PD-L1 or CTLA-4 has limited benefits.Although combination therapies have been proposed to enhance the response rate,severe adverse effects are observed.Thus,alternative immune checkpoints must be identified.The SIGLECs are a family of immunoregulatory receptors(known as glyco-immune checkpoints)discovered in recent years.This review systematically describes the molecular characteristics of the SIGLECs,and discusses recent progress in areas including synthetic ligands,monoclonal antibody inhibitors,and Chimeric antigen receptor T(CAR-T)cells,with a focus on available strategies for blocking the sialylated glycan-SIGLEC axis.Targeting glyco-immune checkpoints can expand the scope of immune checkpoints and provide multiple options for new drug development.
基金Supported by Major Science and Technology Special Project of China Thirteenth Five-Year Plan,No.2018ZX10732401-003-015；Guangxi Key Laboratory for the Prevention and Control of Viral Hepatitis,No.GXCDCKL201901
文摘BACKGROUND Hepatitis B virus (HBV) infection is the primary cause of hepatitis with chronic HBV infection,which may develop into liver fibrosis,cirrhosis and hepatocellular carcinoma.Detection of early-stage fibrosis related to HBV infection is of great clinical significance to block the progression of liver lesion.Direct liver biopsy is regarded as the gold standard to detect and assess fibrosis;however,this method is invasive and prone to clinical sampling error.In order to address these issues,we attempted to find more convenient and effective serum markers for detecting HBV-induced early-stage liver fibrosis.AIM To investigate serum N-glycan profiling related to HBV-induced liver fibrosis and verify multiparameter diagnostic models related to serum N-glycan changes.METHODS N-glycan profiles from the sera of 432 HBV-infected patients with liver fibrosis were analyzed.Significant changed N-glycan levels (peaks)(P <0.05) in differentfibrosis stages were selected in the modeling group,and multiparameter diagnostic models were established based on changed N-glycan levels by logistic regression analysis.The receiver operating characteristic (ROC) curve analysis was performed to evaluate diagnostic efficacy of N-glycans models.These models were then compared with the aspartate aminotransferase to platelet ratio index (APRI),fibrosis index based on the four factors (FIB-4),glutamyltranspeptidase platelet albumin index (S index),GlycoCirrho-test,and GlycoFibro-test.Furthermore,we combined multiparameter diagnostic models with alanine aminotransferase (ALT) and platelet (PLT) tests and compared their diagnostic power.In addition,the diagnostic accuracy of N-glycan models was also verified in the validation group of patients.RESULTS Multiparameter diagnostic models constructed based on N-glycan peak 1,3,4and 8 could distinguish between different stages of liver fibrosis.The area under ROC curves (AUROCs) of Model A and Model B were 0.890 and 0.752,respectively differentiating fibrosis F0-F1 from F2-F4,and F0-F2 from F3-F4,and surpassing other serum panels.However,AUROC (0.747) in Model C used for the diagnosis of F4 from F0-F3 was lower than AUROC (0.795) in FIB-4.In combination with ALT and PLT,the multiparameter models showed better diagnostic power (AUROC=0.912,0.829,0.885,respectively) when compared with other models.In the validation group,the AUROCs of the three combined models (0.929,0.858,and 0.867,respectively) were still satisfactory.We also applied the combined models to distinguish adjacent fibrosis stages of 432patients (F0-F1/F2/F3/F4),and the AUROCs were 0.917,0.720 and 0.785.CONCLUSION Multiparameter models based on serum N-glycans are effective supplementary markers to distinguish between adjacent fibrosis stages of patients caused by HBV,especially in combination with ALT and PLT.
基金This work was supported by the National Natural Science Foundation of China(Nos.22077052,21877052,21907039,22107037)China Postdoctoral Science Foundation(2020M681487,2021M691279)+4 种基金the National Key R&D Program of China(2020YFA0908304)the Natural Science Foundation of Jiangsu Province(BK20180030,BK20190575)the National First-class Discipline Program of Light Industry Technology and Engineering(LITE2018-14)the 111 Project(111-2-06)the Open Project of Key Laboratory of Carbohydrate Chemistry and Biotechnology(Jiangnan University),Ministry of Education(KLCCB-KF202005).
文摘Bacterial surface glycans perform a diverse and important set of biological roles,and have been widely used in the treatment of bacterial infectious diseases.The majority of bacterial surface glycans are decorated with diverse rare functional groups,including amido,acetamidino,carboxamido and pyruvate groups.These functional groups are thought to be important constituents for the biological activities of glycans.Chemical synthesis of glycans bearing these functional groups or their variants is essential for the investigation of structure-activity relationships by a medicinal chemistry approach.To date,a broad choice of synthetic methods is available for targeting the different rare functional groups in bacterial surface glycans.This article reviews the structures of naturally occurring rare functional groups in bacterial surface glycans,and the chemical methods used for installation of these groups.
基金This research was supported by grants from the National Science and Technology Major Project(2018ZX10711-001)the National Natural Science Foundation of China(NSFC)(No.81601813).
文摘Rotavirus(RV)causes acute gastroenteritis in infants and children worldwide.Recent studies showed that glycans such as histo-blood group antigens(HBGAs)function as cell attachment factors affecting RV host susceptibility and prevalence.P[8]is the predominant RV genotype in humans,but the structural basis of how P[8]RVs interact with glycan ligands remains elusive.In this study,we characterized the interactions between P[8]VP8~*s and glycans which showed that VP8~*,the RV glycan binding domain,recognized both mucin core 2 and H type 1 antigens according to the ELISA-based oligosaccharide binding assays.Importantly,we determined the structural basis of P[8]RV-glycans interaction from the crystal structures of a Rotateq P[8]VP8~*in complex with core 2 and H type 1 glycans at 1.82.3?,respectively,revealing a common binding pocket and similar binding mode.Structural and sequence analysis demonstrated that the glycan binding site is conserved among RVs in the P[Ⅱ]genogroup,while genotype-specific amino acid variations determined different glycan binding preference.Our data elucidated the detailed structural basis of the interactions between human P[8]RVs and different host glycan factors,shedding light on RV infection,epidemiology,and development of anti-viral agents.
基金This work was supported by the National Key Research and Development Program of China(2018YFA0507204)the National Natural Sciences Foundation of China(31670165)+1 种基金Wuhan National Biosafety Laboratory,Chinese Academy of Sciences Advanced Customer Cultivation Project(2019ACCP-MS03)the Open Research Fund Program of the State Key Laboratory of Virology of China(2018IOV001).Author Contributions。
文摘Lassa virus(LASV)belongs to the Mammarenavirus genus(family Arenaviridae)and causes severe hemorrhagic fever in humans.The glycoprotein complex(GPC)contains eleven N-linked glycans that play essential roles in GPC functionalities such as cleavage,transport,receptor recognition,epitope shielding,and immune response.We used three mutagenesis strategies(asparagine to glutamine,asparagine to alanine,and serine/tyrosine to alanine mutants)to abolish individual glycan chain on GPC and found that all the three strategies led to cleavage inefficiency on the 2nd(N89),5th(N119),or 8th(N365)glycosylation motif.To evaluate N to Q mutagenesis for further research,it was found that deletion of the 2nd(N89Q)or 8th(N365Q)glycan completely inhibited the transduction efficiency of pseudotyped particles.We further investigated the role of individual glycan on GPC-mediated immune response by DNA immunization of mice.Deletion of the individual 1st(N79Q),3rd(N99Q),5th(N119Q),or 6th(N167Q)glycan significantly enhanced the proportion of effector CD4+cells,whereas deletion of the 1st(N79Q),2nd(N89Q),3rd(N99Q),4th(N109Q),5th(N119Q),6th(N167Q),or 9th(N373Q)glycan enhanced the proportion of CD8+effector T cells.Deletion of specific glycan improves the Th1-type immune response,and abolishment of glycan on GPC generally increases the antibody titer to the glycan-deficient GPC.However,the antibodies from either the mutant or WT GPC-immunized mice show little neutralization effect on wild-type LASV.The glycan residues on GPC provide an immune shield for the virus,and thus represent a target for the design and development of a vaccine.
基金supported by the National Natural Science Foundation of China (Nos. 81672537 and 81470294)the National Science and Technology Major Project of China (No. 2018ZX10302205)Hundred-Talent Program of Shaanxi Province
文摘Sialylation is one of important glycosylation in human beings and plays an important role in cancer development. α2,3-Linked and α2,6-linked sialic acids are normally observed on the end of N-glycans and have different functions. Derivatization on sialic acid was designed to detect the different linkages by MALDI-TOF MS. In this study, a two-step derivatization by dimethylamine and ammonium hydroxide was improved to modify the sialic acid and made it easier to detect the different linkages of sialic acids on MALDI-TOF MS. Using this derivatization method, specific sialic acids linkages on N-glycans of protein samples such as fetuin and lactoferrin were detected. For complex cell samples, increased a2,3-linked and a2,6-linked sialic acids on bi-antennary and tri-antennary N-glycans were observed in A549 cells induced by hypoxia environment. Taken together, our two-step derivatization of sialic acids offers a simple and accurate way to detect specific linkages on N-glycans with MALDI-TOF mass spectrometer.
基金Supported by the National Natural Science Foundation of China(No.30870552)
文摘To explore the effects of traditional herbal medicine Ganoderrna tsugae（G, tsugae） on immunomodulatory and antitumor activities, the crude polysaccharides of G. tsugae were purified by filtration, diethylaminoethyl（DEAE） sepharose-fast flow chromatography and sephadex G-100 size-exclusion chromatography. Two main fractions, pro- tein-containing glyeans CSSLP-1 and CSSLP-2, were obtained via the gradient elution. The protein content, molecu- lar weight, and monosaccharide composition of the two fractions were analyzed. Furthermore, the influence of the protein-containing glycans from G. tsugae on the activation of human acute monocytic leukemia cell line（THP-l） and their antitumor activities to the human hepatocellular liver carcinoma cell（HepG-2） in vitro were evaluated. The re- sults indicate that CSSLP-1 and CSSLP-2 could increase the pinocytie activity of THP-1 cells and induce THP-1 cells to produce the eytokines of TNFa and IL-2, significantly. CSSLP-1 and CSSLP-2 also played an inhibiting effect on the cancer cell（HepG-2）. Moreover, the anti-proliferation activity of CSSLP-1 and CSSLP-2 increased with the par- ticipation of TNFa and IL-2 or other antitumor factors induced from THP-1 cells by G. tsugae protein-containing glycan fractions.
基金the support from the National Natural Science Foundation of China (Nos.21974067, 22274073, 81971373 and 82001535)the National Key Research and Development Program of China (No.2018YFC1004700)+1 种基金Fundamental Research Funds for the Central Universities (Nos.020514380309,021414380502 and 2022300324)the State Key Laboratory of Analytical Chemistry for Life Science (Nos.5431ZZXM2305 and 5431ZZXM2204)。
文摘Spermatogenesis, maturation, capacitation and fertilization are precisely regulated by glycosylation. However, the relationship between altered glycosylation patterns and the onset and development of reproductive disorders is unclear, mainly limited by the lack of in situ imaging techniques for spermatozoa glycosylation. We developed an efficient and highly specific spermatozoa glycan imaging technique based on the robust chemoselective labeling of sialic acid(Sia) and N-acetyl-D-galactosamine(Gal/GalNAc). We further proposed a “tandem glycan chemoselective labeling” strategy to achieve simultaneous imaging of two types of glycans on spermatozoa. We applied the developed method to the spermatozoa from oligozoospermic patients and diabetic mice and found that these spermatozoa showed higher levels of Sia and Gal/Gal NAc expression than the normal groups. Moreover, spermatozoa from diabetic mice showed a severe decrease in number, viability, and forward motility, suggesting that in vivo glucose metabolism disorders may lead to an elevated level of spermatozoa glycosylation and have a correlation with the development of oligoasthenotspermia. Our work provides a research tool to reveal the relationship between glycosylation modification and spermatozoa quality, and a promising clue for the development of glycan-based reproductive markers.
基金supported by Basic Science Research Program through the National Research Foundation of Korea(NRF)funded by the Ministry of Education(NRF-RS-2023-00275307)。
文摘Background Optimal gut health is important to maximize growth performance and feed efficiency in broiler chickens.A total of 1,365 one-day-old male Ross 308 broiler chickens were randomly divided into 5 treatments groups with 21 replicates,13 birds per replicate.The present research investigated effects of microbial muramidase or a precision glycan alone or in combination on growth performance,apparent total tract digestibility,total blood carotenoid content,intestinal villus length,meat quality and gut microbiota in broiler chickens.Treatments included:NC:negative control(basal diet group);PC:positive control(basal diet+0.02%probiotics);MR:basal diet+0.035%microbial muramidase;PG:basal diet+0.1%precision glycan;and MRPG:basal diet+0.025%MR+0.1%PG,respectively.Results MRPG group increased the body weight gain and feed intake(P<0.05)compared with NC group.Moreover,it significantly increased total serum carotenoid(P<0.05)and MRPG altered the microbial diversity in ileum contents.The MRPG treatment group increased the abundance of the phylum Firmicutes,and family Lachnospiraceae,Ruminococcaceae,Oscillospiraceae,Lactobacillaceae,Peptostreptococcaceae and decreased the abundance of the phylum Campilobacterota,Bacteroidota and family Bacteroidaceae.Compared with the NC group,the chickens fed MRPG showed significantly increased in duodenum villus length at end the trial.Conclusion In this study,overall results showed that the synergetic effects of MR and PG showed enhancing growth performance,total serum carotenoid level and altering gut microbiota composition of broilers.The current research indicates that co-supplementation of MR and PG in broiler diets enhances intestinal health,consequently leading to an increased broiler production.
基金Supported by the National Natural Science Foundation of China(30800193)Grant from Centre for International Mobility(CIMO),Finland
文摘Native and methyl-esterified sialylated glycans were analyzed with 2,4,6-trihydroxyacetophenone（THAP）and 2,5-dihydroxybenzoic acid（DHB）as matrix by a matrix-assisted laser desorption/ionization time-of-flight mass spectrometer（MALDI-TOF MS）.High quality negative-ion spectra of commercial sialylated glycan were obtained with THAP as matrix.Detection limit of the glycan was less than 0.1 pmol.After methyl esterification of sialic acid（SA）residue,sialylated glycans were detected sensitively in the positive-ion mode using DHB as matrix.Neutral and sialylated glycans from the mixture of asialofetuin and fetuin were methylesterified and simultaneously recognized in one manipulation.Methyl esterification of SA residue offers a convenient and sensitive way to identify the structure of N-linked glycans for glycan profiling.
基金The Chinese Academy of Sciences:KFJ-BRP-007-019,the authors declare no competing interests.
文摘As the two most principal active substances in the corn silk,polysaccharides and flavonoids,the mechanism of interaction between them has been a topic of intense research.This study provides an in-depth investigation of the interaction mechanism between corn silk glycans and luteoloside(LUT)and the synergistic role that result from this interaction.The interaction mechanism was evaluated by isothermal titration calorimetry(ITC)and circular dichroism(CD),and the synergistic role was evaluated by the expression of glucose transporters(GLUT-1),insulin secretion and surface plasmon resonance(SPR).CD and ITC results indicated that the interaction between CSGs and LUT mainly driven by the Cotton effects,enthalpy and entropy-driven.This interaction precipitated the formation of complexes(CSGs/LUT complexes)between corn silk glycans(CSGs)with four different molecular weights and luteoloside(LUT).Furthermore,the CSGs and LUT play a synergistic role in glucose regulation through GLUT-1 expression and insulin secretion experiments,compared to single luteoloside group.
基金the National Natural Science Foundation of China (81473179 and 81673388)Priority Academic Program Development of Jiangsu Higher Education Institutions (PAPD, YX13200111)the funding for Jiangsu Key Laboratory of Translational Research and Therapy for Neuro-PsychoDiseases (BM2013003)
文摘The glycosylation of proteins is responsible for their structural and functional roles in many cellular activities.This work describes a strategy that combines an efficient release, labeling and liquid chromatography–mass spectral analysis with the use of a comprehensive database to analyze N-glycans. The analytical method described relies on a recently commercialized kit in which quick deglycosylation is followed by rapid labeling and cleanup of labeled glycans. This greatly improves the separation, mass spectrometry（MS） analysis and fluorescence detection of N-glycans. A hypothetical database, constructed using Glyc Resoft, provides all compositional possibilities of N-glycans based on the common sugar residues found in N-glycans. In the initial version this database contains ＆gt; 8,700 N-glycans, and is compatible with MS instrument software and expandable. N-glycans from four different well-studied glycoproteins were analyzed by this strategy. The results provided much more accurate and comprehensive data than that had been previously reported. This strategy was then used to analyze the N-glycans present on the membrane glycoproteins of gastric carcinoma cells with different degrees of differentiation. Accurate and comprehensive N-glycan data from those cells was obtained efficiently and their differences were compared corresponding to their differentiation states. Thus, the novel strategy developed greatly improves accuracy, efficiency and comprehensiveness of N-glycan analysis.
基金supported by National Natural Science Foundation of China(Grant Nos.92351001,31970170,and 31661143033)PI Project of Southern Marine Science and Engineering Guangdong Laboratory(Guangzhou)(GML20240002).
文摘Impact statement.Spindle-shaped viruses exclusively infect archaea.Fuselloviruses represent a large group of spindle-shaped viruses and infect hyperthermophilic archaea of the order Sulfolobales.Although the first fusellovirus was identified nearly 40 years ago,the mechanism of host infection by these viruses remains poorly understood.Here,we show that SSV19,a fusellovirus isolated from a hot spring in the Philippines,is capable of hydrolyzing the host cell surface glycan identified as a heptasaccharide chain of QuiS_(1)Hex_(4)HexNAc_(2).Our findings provide significant insights into the molecular strategy of host recognition and,possibly,entry by an archaeal virus.
基金supported by grants from the Key Technologies Research and Development Program(2022YFC3400900)Postdoctoral Fellowship Program of CPSF(GZB20230886)+3 种基金China Postdoctoral Science Foundation(2023M743998)National Natural Science Foundation of China(82030046)the Program for Guangdong Introducing Innovative and Entrepreneurial Teams(2019BT02Y198)Guangdong Science and Technology Department(2020B1212030004).
文摘Coronaviruses are enveloped RNA viruses distinguished by their crown-like surface spikes. These viruses cause various diseases in humans and animals, including respiratory, gastrointestinal, and neurological disorders. The health risks posed by coronaviruses are substantial, as demonstrated by severe acute respiratory syndrome-coronavirus (SARS-CoV) and SARS-CoV-2. SARS-CoV triggered a global outbreak of SARS in 2003, and SARS-CoV-2 has been responsible for the ongoing coronavirus disease 2019 (COVID-19) pandemic since the end of 2019. Furthermore, four human coronaviruses (HCoVs), HCoV-229E, HCoV-OC43, HCoV-NL63, and HCoV-HKU1, have been implicated in 15% to 30% cases of the common cold.[1] Given their prevalence and potential to cause various diseases, understanding the mechanisms underlying the invasion into cells of the coronavirus is essential for public health. By binding to specific cell receptors, the spike protein of coronaviruses undergoes conformational changes that promote membrane fusion and entry of the virus into host cells. Although significant progress has been made in elucidating the mechanisms of coronavirus entry, the detailed molecular interactions between viral proteins and host receptors remain incompletely understood.
基金supported by the National Natural Science Foundation of China(32271521,31971361,and 82170763)the State Key Research Development Program of China(2022YFC2603902)+2 种基金the Natural Science Foundation of Beijing Municipality(5212013)the Fundamental Research Funds for the Central Universities(PT2406)the Talent Cultivation of State Key Laboratory of Organic-Inorganic Composites,Beijing University of Chemical Technology(oic-2024020007)。
文摘Fluorescent DNA probes commonly used for in situ imaging of cell surface glycans are“always-on”probes(AOPs),which produce high background noise and lack spatial specificity.Molecular dynamics simulations indicate that single-stranded DNA(ssDNA)can stably associate with the cell membrane,contributing to persistent background signals in AOPs.To overcome this challenge,a stepwise activated probe(SAP)is developed for in situ imaging of cell surface glycans.In SAP,the cell surface glycan is labeled by rolling circle amplification product through metabolic glycan labeling strategies.The product hybridizes a fluorophore and quencher-labeled ssDNA reporter to form a double strand producing primary enhancement of fluorescence signal.Nicking enzyme(Nb.BbvCI)cleaves the double strand and releases the quencher,further enhancing the signal.Nonspecific absorption of the reporter on the cellular membrane does not increase the fluorophore-quencher distance in SAP or trigger Nb.BbvCI cleavage.As a result,only the glycan sites are illuminated.SAP not only exhibits high imaging specificity but also greatly simplifies the imaging procedure by reducing the washing steps.SAP offers a considerable improvement in detection specificity and sensitivity by employing a two-step activation and amplification process,making it a powerful method for in situ fluorescence imaging.
基金supported by grants from the National Key Research and Development Program of China(2022YFC3401500 and 2020YFA0803201 to P.W.)the Major Research Plan of the National Natural Science Foundation of China(Grant No.92153301 to L.Lin.)+2 种基金the National Natural Science Foundation of China(Grant No.22177126 to L.Lin.)the National Natural Science Foundation of China(82341028,31920103007 to P.W.)the Key R&D Projects in Ningxia Hui Autonomous Region(2021BFH03001).
文摘Among the leading methods for triggering therapeutic anti-cancer immunity is the inhibition of immune checkpoint pathways.N-glycosylation is found to be essential for the function of various immune checkpoint proteins,playing a critical role in their stability and interaction with immune cells.Removing the N-glycans of these proteins seems to be an alternative therapy,but there is a lack of a de-N-glycosylation technique for target protein specificity,which limits its clinical application.Here,we developed a novel technique for specifically removing N-glycans from a target protein on the cell surface,named deglycosylation targeting chimera(DGlyTAC),which employs a fusing protein consisting of Peptide-N-glycosidase F(PNGF)and target-specific nanobody/affibody(Nb/Af).The DGlyTAC technique was developed to target a range of glycosylated surface proteins,especially these immune checkpoints—CD24,CD47,and PD-L1,which minimally affected the overall N-glycosylation landscape and the N-glycosylation of other representative membrane proteins,ensuring high specificity and minimal off-target effects.Importantly,DGlyTAC technique was successfully applied to lead inactivation of these immune checkpoints,especially PD-L1,and showed more potential in cancer immunotherapy than inhibitors.Finally,PD-L1 targeted DGlyTAC showed therapeutic effects on several tumors in vivo,even better than PD-L1 antibody.Overall,we created a novel target-specific N-glysocylation erasing technique that establishes a modular strategy for directing membrane proteins inactivation,with broad implications on tumor immune therapeutics.
基金supported by grants from SSP:co-funded by the European Union(ERC,GlycanSwitch,101071386).Views and opinions expressed are however those of the author(s)only and do not necessarily reflect those of the European Union or the European Research Council Executive Agency.Neither the European Union nor the granting authority can be held responsible for them.The work was also co-funded by EU GlycanTrigger-grant Agreement No:101093997.Views and opinions expressed are however those of the author(s)only and do not necessarily reflect those of the European Union or European Health and Digital Executive Agency.Neither the European Union nor the granting authority can be held responsible for them.SSP also acknowledges funding by“2022 LRA Lupus Innovation Award”and by“European Crohn’s and Colitis Organisation(ECCO)Pioneer Award 2021”.SSP also acknowledges the US Department of Defense,US Army Medical Research Acquisition Activity,FY18 Peer Reviewed Medical Research Program Investigator-Initiated Research Award(award number W81XWH1920053)as well as grant funded by the Portuguese Foundation for Science and Technology–FCT(EXPL/MED-ONC/0496/2021).IA acknowledges FCT for funding(2022.00337.CEECIND).JG acknowledges funding from ESCMID(ESCMID Research Grant 2022),ECCO(ECCO Grant 2023)and FCT(2020.00088.CEECIND).G.A.R acknowledges grants from the Argentinean Agency for Promotion of Science,Technology and Innovation(PICT 2017-0494,PICT-FBB 620 and PICT 2020-01552).The authors are also thankful for generous support from Sales(Argentina),Bunge&Born(Argentina),Baron(Argentina),Williams(Argentina)and Richard Lounsbery(USA)Foundations,as well as donations from Ferioli-Ostry and Caraballo families to GAR.
文摘The immune system is coordinated by an intricate network of stimulatory and inhibitory circuits that regulate host responses against endogenous and exogenous insults.Disruption of these safeguard and homeostatic mechanisms can lead to unpredictable inflammatory and autoimmune responses,whereas deficiency of immune stimulatory pathways may orchestrate immunosuppressive programs that contribute to perpetuate chronic infections,but also influence cancer development and progression.Glycans have emerged as essential components of homeostatic circuits,acting as fine-tuners of immunological responses and potential molecular targets for manipulation of immune tolerance and activation in a wide range of pathologic settings.Cell surface glycans,present in cells,tissues and the extracellular matrix,have been proposed to serve as“self-associated molecular patterns”that store structurally relevant biological data.The responsibility of deciphering this information relies on different families of glycan-binding proteins(including galectins,siglecs and C-type lectins)which,upon recognition of specific carbohydrate structures,can recalibrate the magnitude,nature and fate of immune responses.This process is tightly regulated by the diversity of glycan structures and the establishment of multivalent interactions on cell surface receptors and the extracellular matrix.Here we review the spatiotemporal regulation of selected glycan-modifying processes including mannosylation,complex N-glycan branching,core 2 O-glycan elongation,LacNAc extension,as well as terminal sialylation and fucosylation.Moreover,we illustrate examples that highlight the contribution of these processes to the control of immune responses and their integration with canonical tolerogenic pathways.Finally,we discuss the power of glycans and glycan-binding proteins as a source of immunomodulatory signals that could be leveraged for the treatment of autoimmune inflammation and chronic infection.
基金supported by the National Natural Science Foundation of China(91853120)the National Major Scientific and Technological Special Project of China(2018ZX09711001-013 and 2018ZX09711001-005)+2 种基金the National Key Research and Development Program of China(2018YFE0111400 and 2016YFD0500300)the State Key Laboratory of Bioactive Substance and Function of Natural Medicines,Institute of Materia Medica,the Chinese Academy of Medical Sciences and Peking Union Medical College,the NIH Research Project Grant Program(R01 EB025892)the CRP-ICGEB Research Grant 2019(CRP/CHN19-02)。
文摘The pandemic of the severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)has caused a high number of deaths in the world.To combat it,it is necessary to develop a better understanding of how the virus infects host cells.Infection normally starts with the attachment of the virus to cell-surface glycans like heparan sulfate(HS)and sialic acid-containing glycolipids/glycoproteins.In this study,we examined and compared the binding of the subunits and spike(S)proteins of SARS-CoV-2,SARS-Co V,and Middle East respiratory disease(MERS)-Co V to these glycans.Our results revealed that the S proteins and subunits can bind to HS in a sulfation-dependent manner and no binding with sialic acid residues was detected.Overall,this work suggests that HS binding may be a general mechanism for the attachment of these coronaviruses to host cells,and supports the potential importance of HS in infection and in the development of antiviral agents against these viruses.
