Glycomed试图从Lilly手中重新获得抗再狭窄药物的销售权

Glycomed Inc.(Alameda,CA)在公布其药物 Astenose 对血管成形术引起的血管再狭窄有越来越明显的预防作用后不久,就开始与其研究和开发该药的合作伙伴 Eli Lilly & Co.(Indianapolis,IN)进行辩论,以期重新获得对该药的销售权。再狭窄是由于经过血管成形术或分流术等外科手术后平滑肌细胞快速增生所引起的动脉慢性再闭合。Glycomed 最近发表的活体试验数据表明,将其产品(Astenose)注射到经血管成形术后的家兔体内时,

N-glycan biosignatures as a potential diagnostic biomarker for earlystage pancreatic cancer

BACKGROUND Pancreatic ductal adenocarcinoma(PDAC)has a poor prognosis,with a 5-year survival rate of less than 10%,owing to its late-stage diagnosis.Early detection of pancreatic cancer(PC)can significantly increase survival rates.AIM To identify the serum biomarker signatures associated with early-stage PDAC by serum N-glycan analysis.METHODS An extensive patient cohort was used to determine a biomarker signature,in-cluding patients with PDAC that was well-defined at an early stage(stages I and II).The biomarker signature was derived from a case-control study using a case-cohort design consisting of 29 patients with stage I,22 with stage II,4 with stage III,16 with stage IV PDAC,and 88 controls.We used multiparametric analysis to identify early-stage PDAC N-glycan signatures and developed an N-glycan sig-nature-based diagnosis model called the“Glyco-model”.RESULTS The biomarker signature was created to discriminate samples derived from patients with PC from those of controls,with a receiver operating characteristic area under the curve of 0.86.In addition,the biomarker signature combined with cancer antigen 19-9 could discriminate patients with PDAC from controls,with a receiver operating characteristic area under the curve of 0.919.Glyco-model demonstrated favorable diagnostic performance in all stages of PC.The diagnostic sensitivity for stage I PDAC was 89.66%.Core Tip:This study employed a patient cohort to investigate the N-glycan signature of early-stage pancreatic cancer(PC).Serum N-glycans analysis was conducted to identify the serum biomarker signature associated with early-stage pancreatic ductal adenocarcinoma(PDAC),resulting in the identification of nine early-stage PDAC N-glycan signatures.Subsequently,utilizing these biosignatures,a diagnostic model named the“Glyco-model”was developed,demonstrating promising diagnostic performance across all stages of PC.The study revealed that the diagnostic sensitivity for stage I PDAC was determined to be 89.66%.Consequently,this diagnostic model exhibits potential as a prospective strategy for the early detection of PDAC.

Detecting cancer cell-surface glycome profilings by lectin microarray

Objective: Glycomics is an emerging field in the postgenome and postproteome era. Glycosylation changes of cell-surface have been observed in tumour cell progression

人类蛋白质N-糖基化的十二年全基因组关联研究

Most human-secreted and membrane-bound proteins have covalently attached oligosaccharide chains or glycans.Glycosylation influences the physical and chemical properties of proteins,as well as their biological functions.Unsurprisingly,alterations in protein glycosylation have been implicated in a growing number of human diseases,and glycans are increasingly being considered as potential therapeutic targets,an essential part of therapeutics,and biomarkers.Although glycosylation pathways are biochemically well-studied,little is known about the networks of genes that guide the cell-and tissue-specific regulation of these biochemical reactions in humans in vivo.The lack of a detailed understanding of the mechanisms regulating glycome variation and linking the glycome to human health and disease is slowing progress in clinical applications of human glycobiology.Two of the tools that can provide much sought-after knowledge of human in vivo glycobiology are human genetics and genomics,which offer a powerful data-driven agnostic approach for dissecting the biology of complex traits.This review summarizes the current state of human populational glycogenomics.In Section 1,we provide a brief overview of the N-glycan’s structural organization,and in Section 2,we give a description of the major blood plasma glycoproteins.Next,in Section 3,we summarize,systemize,and generalize the results from current N-glycosylation genome-wide association studies(GWASs)that provide novel knowledge of the genetic regulation of the populational variation of glycosylation.Until now,such studies have been limited to an analysis of the human blood plasma N-glycome and the N-glycosylation of immunoglobulin G and transferrin.While these three glycomes make up a rather limited set compared with the enormous multitude of glycomes of different tissues and glycoproteins,the study of these three does allow for powerful analysis and generalization.Finally,in Section 4,we turn to genes in the established loci,paying particular attention to genes with strong support in Section 5.At the end of the review,in Sections 6 and 7,we describe special cases of interest in light of new discoveries,focusing on possible mechanisms of action and biological targets of genetic variation that have been implicated in human protein N-glycosylation.

Cloning,Expressing and Characterizing a Novel Cold-Adapted Laminarinase from Marine Bacterium Wenyingzhuangia aestuarii OF219

Laminarinases reveal potential application in the field of food and biotechnology.In this research,a novel GH16 family laminarinase,designated as Lam16A_Wa,was cloned from the genome of marine bacterium Wenyingzhuangia aestuarii OF219 and expressed in Escherichia coli.Lam16A_Wa demonstrates a relatively low optimal reaction temperature(35℃)and a cold-adapted feature.Its optimal pH value is 6.0 and is stable in a broad pH range from 3.0 to 11.0.A glycomics strategy was employed to investigate the hydrolytic pattern of Lam16A_Wa.The enzyme was confirmed as a random endo-acting glycoside hydrolase.Its minimum substrate was laminarin pentasaccharide,and the major final products are oligosaccharides,including disaccharide to pentasaccharide.The Lam16A_Wa provides a novel and well-defined tool for the molecular tailoring of laminarin.

Possible role of sialylation of retinal protein glycans in the regulation of electroretinogram response in mice

AIM： To evaluate if the nature, degree and extent of Siaa2-3-/Siaa2-6-sialylation of retinal protein glycans plays a possible role in the development and regulation of electroretinogram response （ERG） in mice. METHODS： Proteins extracted, from retinae of postnatal day 2 （PN2）, PN7, and PN14 wild type （wt） and retinal degeneration 1 （rdl） mice were quantified, labeled and used for lectin-microarray profiling with immobilized lectins which recognize a wide range of N-/O-glycans. Net fluorescence intensities of lectin-ligand complexes were measured and images of fluorescent lectin-microarrays were acquired. From the binding curves between each lectin and protein extracts from PN14 wt and PN14 rdl mice retinae, the protein concentration was selected to determine optimum signal intensity for lectin-ligand binding. Mean_＋SEM values of proteins and fluorescence- intensities of lectin-ligand-complexes between 45 lectins and 36 protein extracts from wt and rdl mice retinae were compared for significance of differences. RESULTS： Comparison of the progressive relative changes in the sialylated glycans of retinal proteins from wt and rdl mice showed that Siao2-3Gall-4GIcNAc-glycans （but not Siaa2-6-glycans） were detectable and quantifiable from the retinal-proteins of PN7 and PN14 wt and rdl mice. Siaa2- 3-sialylation of retinal-protein Gal/o-linked-Gal-glycans was significantly increased with age in PN7 and PN14 wt and less so in PN14 rdl mice. Siao2-3-/Siaa2-6-sialylation of retinal-protein Gal/a-linked-Gal-glycans was absent in PN2 wt and rdl mice. Comparison of published ERG responses of wt and rdl mice retinae with degree of Siaa2- 3-sialylation of retinal-protein-glycans showed that PN2 wt and rdl mice lack both the ERG response and Siaa2- 3-/Siao2-6-sialylation of retinal-protein Gal/a-linked-Gal-glycans; rdl mice with relatively lower Siaa2-3-sialylation of retinal-protein Gal/a-linked-Gal-glycans showed aberrant ERG response; and wt mice with significantly higher Siaa2-3-sialylation of retinal-protein Gal/a-linked- Gal-glycans showed normal ERG response. CONCLUSION： Degree of Siaa2-3-sialylation of giycans possibly regulates ERG function in mice.

Single-cell omics analyses with single molecular detection:challenges and perspectives

The ultimate goal of single-cell analyses is to obtain the biomolecular content for each cell in unicellular and multicellular organisms at different points of their life cycle under variable environmental conditions.These require an assessment of:a)the total number of cells,b)the total number of cell types,and c)the complete and quantitative single molecular detection and identification for all classes of biopolymers,and organic and inorganic compounds,in each individual cell.For proteins,glycans,lipids,and metabolites,whose sequences cannot be amplified by copying as in the case of nucleic acids,the detection limit by mass spectrometry is about 105 molecules.Therefore,proteomic,glycomic,lipidomic,and metabolomic analyses do not yet permit the assembly of the complete single-cell omes.The construction of novel nanoelectrophoretic arrays and nano in microarrays on a single 1-cm-diameter chip has shown proof of concept for a high throughput platform for parallel processing of thousands of individual cells.Combined with dynamic secondary ion mass spectrometry,with 3D scanning capability and lateral resolution of 50 nm,the sensitivity of single molecular quantification and identification for all classes of biomolecules could be reached.Further development and routine application of such technological and instrumentation solution would allow assembly of complete omes with a quantitative assessment of structural and functional cellular diversity at the molecular level.

Updated clinical and glycomic features of mannosyl-oligosaccharide glucosidase deficiency:Two case reports

BACKGROUND Mannosyl-oligosaccharide glucosidase(MOGS)deficiency is an extremely rare type of congenital disorder of glycosylation(CDG),with only 12 reported cases.Its clinical,genetic,and glycomic features are still expanding.Our aim is to update the novel clinical and glycosylation features of 2 previously reported patients with MOGS-CDG.CASE SUMMARY We collected comprehensive clinical information,and conducted the immunoglobulin G1 glycosylation assay using nano-electrospray ionization source quadruple time-of-flight mass spectrometry.Novel dysmorphic features included an enlarged tongue,forwardly rotated earlobes,a birth mark,overlapped toes,and abnormal fat distribution.Novel imaging findings included pericardial effusion,a deep interarytenoid groove,mild congenital subglottic stenosis,and laryngomalacia.Novel laboratory findings included peripheral leukocytosis with neutrophil predominance,elevated C-reactive protein and creatine kinase,dyslipidemia,coagulopathy,complement 3 and complement 4 deficiencies,decreased proportions of T lymphocytes and natural killer cells,and increased serum interleukin 6.Glycosylation studies showed a significant increase of hypermannosylated glycopeptides(Glc3Man7GlcNAc2/N2H10 and Man5GlcNAc2/N2H5)and hypersialylated glycopeptides.A compensatory glycosylation pathway leading to an increase in Man5GlcNAc2/N2H5 was indicated with the glycosylation profile.CONCLUSION We confirmed abnormal glycomics in 1 patient,expanding the clinical and glycomic spectrum of MOGS-CDG.We also postulated a compensatory glycosylation pathway,leading to a possible serum biomarker for future diagnosis.

Insights into glycan biosynthesis in chemically-induced hepatocellular carcinoma in rats: A glycomic analysis

AIM： To evaluate the qualitative and quantitativechanges in N-linked glycosylation, which occurredin association with diethyl nitrosamine-inducedhepatocellular carcinoma （HCC） in rodents.METHODS： Liver tissues of （1） normal （non-tumorbearing）rats; and （2） tumor-bearing rats; were collectedand were used for histological and GlycanMap？ analyses.Briefly, GlycanMap？ analysis is a high-throughputassay that provides a structural and quantitativereadout of protein-associated glycans using a unique,automated 96-well assay technology coupled tomatrix-assisted laser desorption/ionization time-offlightmass spectrometry and custom bioinformatics.Histopathological studies were carried out to ensure thedevelopment of HCC in the tested animals.RESULTS： The N-glycomic analysis revealed 5glycans; Glc1Man9GlcNAc2, Gal2Man3GlcNac4Fuc1Neu1,Man4G l c N a c 2, G a l 2Man3G l c N a c 4Neu3OAc 3, andMan3GlcNac5Fuc1, which showed significant changesin rat HCC tissues when compared with normal livertissues. Four glycans were increased （P 〈 0.05） andGlc1Man9GlcNAc2 was decreased （5.89 ± 0.45 vs 3.54± 0.21, P 〈 0.01） in HCC tissues compared to normal liver tissues. An increase （66.5 ± 1.05 vs 62.7 ± 1.1,P 〈 0.05） in high-mannose structures in HCC rats wasobserved compared to normal rats. Importantly, HCCrats showed an increase （P 〈 0.05） in both tumorassociatedcarbohydrates and in branched glycans. Thechanges in glycans correlated well with glycan flowchanges reported in the glycan biosynthetic pathway,which indicates the importance of enzyme activitiesinvolved in glycan synthesis at different subcellularlocalizations.CONCLUSION： The reported HCC-associated changesin glycan flow and subcellular localization explain theincrease in high mannose glycans and siayl Lewisglycans common in HCC liver tissues.

N-glycans released from glycoproteins using a commercial kit and comprehensively analyzed with a hypothetical database

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.

Control of lupus activity during pregnancy via the engagement of IgG sialylation:novel crosstalk between IgG sialylation and pDC functions

Immunoglobulin(IgG)glycosylation affects the effector functions of IgG in a myriad of biological processes and has been closely associated with numerous autoimmune diseases,including systemic lupus erythematosus(SLE),thus underlining the pathogenic role of glycosylation aberration in autoimmunity.This study aims to explore the relationship between IgG sialylation patterns and lupus pregnancy.Relative to that in serum samples from the control cohort,IgG sialylation level was aberrantly downregulated in serum samples from the SLE cohort at four stages(from preconception to the third trimester of pregnancy)and was significantly associated with lupus activity and fetal loss during lupus pregnancy.The type I interferon signature of pregnant patients with SLE was negatively correlated with the level of IgG sialylation.The lack of sialylation dampened the ability of IgG to suppress the functions of plasmacytoid dendritic cells(pDCs).RNA-seq analysis further revealed that the expression of genes associated with the spleen tyrosine kinase(SYK)signaling pathway significantly differed between IgG-and deSia-IgG-treated pDCs.This finding was confirmed by the attenuation of the ability to phosphorylate SYK and BLNK in deSia-IgG.Finally,the coculture of pDCs isolated from pregnant patients with SLE with IgG/deSia-IgG demonstrated the sialylation-dependent anti-inflammatory function of IgG.Our findings suggested that IgG influences lupus activity through regulating pDCs function via the modulation of the SYK pathway in a sialic acid-dependent manner.

Mass spectrometry analysis of melanoma related O-glycans in sera

Glycomics comparison was carried out by screening melanoma serum biomarkers between C57 mice with and without B16 implanted. O-glycans were released from 10 μL sera by β-elimination, purified by Graphitized Carbon Cartridge Solid Phase Extraction (GCC-SPE) and analyzed by MALDI-QIT-TOF-MS. MS raw data were acquired and exported by Launchpad software. MATLAB was then applied for further data analysis. 10 Glycans were considered to have stable changes after B16 implantation and 5 of them were under structural analysis via MS/MS.