3^rd Glycobiology World Congress June 26-28, 2017 London, UK

Jorge Justino has his expertise in the biological evaluation and toxicity studies of natural and synthetic compounds and food chemistry. Full Professor of Escola Superior Agrária de Santarém, MSc. and Ph.D in Chemical Engineering at Instituto Superior Técnico – Universidade Técnica de Lisboa, Habilitation in Food Chemistry at Universidade de Trás-Os-Montes e Alto Douro. He is President of Instituto Politécnico de Santarém and former Director of Escola Superior Agrária de Santarém, He was a member of the networks Euroglycoforum and Prion Chemical Biology Network and is the representative member of the Instituto Politécnico de Santarém at the European Innovation Partnership A3 Group FCUL consortium.

The emergence of antibiotic resistance encouraged chemists to explore new structures as new antibiotics with new modes of action. Dodecyl deoxy glycoside structures have shown a potent activity against Bacillus spp, in particular B. cereus.1-3 A small library of related glycosides, differing in saccharide configuration and deoxygenation pattern, as well as aglycone structure have been synthesized for the recognition of the structural features that determine the selectivity for Bacillus cereus. Their preparation was accomplished by reaction of glycals with alcohols, catalyzed by triphenylphosphane hydrobromide.4 Depending on their structural features, some of these compounds demonstrated a potent activity against B. cereus. In addition, they showed low/very low toxicity and are not genotoxic. This work clearly demonstrates the uniqueness of carbohydrates which stereochemistry and chemical structure can tune the bioactivity exhibited by stereoisomers.

Nora Berois is Medical Doctor, Gynaecologist, and completed her PhD at the Technological University of Compiègne, France. She is Associated Investigator at the Laboratory of Glycobiology and Tumor Immunology at Pasteur Institut, Montevideo, Uruguay. Her major research interest is biomarkers in precision medicine. She has published 22 papers in reputed journals.

Lung cancer is currently the leading cause of cancer-related death worldwide, accounting for approximately a third of all cancer diagnoses and related-deaths. Non–small cell lung cancer (NSCLC) represents nearly 80% of lung tumors; the two most common NSCLC histological types are squamous cell carcinoma (SCC) and adenocarcinoma (ADCA). Several diagnosis procedures detecting altered glycosylation have been developed and incorporated as assistant procedures in clinical oncology. The synthesis of mucin-type O-glycans is started in a reaction catalyzed by UDP-N-acetyl-D-galactosamine:polypeptide N-acetylgalactosaminyltransferases (GalNAc-Ts). This is a complex family of enzymes of at least 20 members in humans. We previously found that GALNT13, the gene encoding GalNAc-T13 isoenzyme, is a strong predictor of poor clinical outcome in neuroblastoma patients (Berois et al., Clin Chem. 2006; 52:1701-12). In the present study, we evaluated GalNAc-T13 expression in human NSCLC. We produced a monoclonal antibody (MAb T13.5) that was used to assess the expression profile of the GalNAc-T13 protein in a well-defined population of 443 surgically resected NSCLC patients with 7 years of follow-up. We found that ADCAs expressed higher levels of the enzyme than SCCs. GalNAc-T13 expression correlated significantly with worse overall survival in ADCA patients treated with neoadjuvant chemotherapy. These data suggest that GalNAc-T13 could be a novel marker associated to chemoresistance in lung adenocarcinomas.

Jonathan Cawley is currently a fourth year PhD student in the School of Biotechnology at Dublin City University. He received his B.Sc. in Biotechnology (1st class honours) from DCU in 2013. His research focuses on the glycosylation changes to the CHO cell surface by probing cells with recombinant lectins, which he has expressed and purified, and commercial lectins. Subsequently the cells are analysed with a fluorescent microscope and flow cytometer.

Chinese hamster ovary (CHO) cells are extensively used for the production of biopharmaceutical products. In 1985 the first recombinant proteins (i.e., monoclonal antibodies, blood factors, growth hormones and cytokines etc.) are produced in CHO cells. Proteins produced in CHO frequently undergo post-translational modifications (PTM). The most common PTM is glycosylation which can alter the function, stability, immunogenicity and efficacy of the resulting protein. Changes to the CHO glycocalyx, carbohydrate coat surrounding the cell membrane, may be informative to changes on the product being produced. It is therefore of great interest to understand how the host cell changes in different culture conditions. Fluorescent microscopy and flow cytometry are employed to perform glycoanalysis on CHO DP-12 cells. Recombinant eukaryotic lectins, purified using immobilized metal affinity chromatography (IMAC), and commercial plant and fungal lectins are used to probe CHO cells in order to establish a lectin binding profile. Lectins from non-plant sources are perfect glycan probes as they are generally nontoxic but they are also superior to other binding proteins such as antibodies whose specificities for carbohydrate targets are ill-defined. In this work a recombinant eukaryotic lectin, Agrocybe aegerita lectin 2 (AAL-2), was expressed, purified and characterised. It was then successfully used to probe the CHO DP-12 surface, along with other lectins, demonstrating that a novel lectin produced recombinantly is the optimal choice of probe for investigating the cell surface glycosylation of live cells.

Dr. Wang has completed her PhD in 2004 in Taiwan and postdoctoral studies in Max-Planck Institute of Biochemistry, Germany. She was a project leader of University of Basel (2009-2010) and now an associate professor of National Tsing Hua University in Taiwan. Her expertise involves time-lapse live cell microscopy and the application this technology to monitor timely changes of cells upon physiological and pathological conditions.

Glycosaminoglycans (GAGs) are carbohydrates that are usually attached to extracellular or plasma membrane proteins to form proteoglycans. A specific GAG-binding peptide (GBP-HH) has been identified from core heparan sulfate binding region in human RNase3, a unique RNase family member with in vitro antitumor activity. GBP-HH displayed a strong cell surface binding and penetrating activity with no obvious effect on general cell proliferation in a variety of cancer cell lines. Here we show that GBP-HH significantly reduced cell migration in A549 lung cancer and OECM-1 oral cancer cells, suggesting that GBP had an inhibitory effect on cancer cell motility. Specifically, GBP-HH also blocked HB-EGF-induced cell migration, but not EGF. This implied that GBP-HH might control cell migration through its heparin sulfate binding ability. We found that treatment with GBP-HH increased the overall number of focal adhesion in A549 cell, as well as increased length of focal adhesion complex. To further study the dynamics of focal adhesion, A549 cells were transfected with vinculin-GFP expression constructs and subjected to time-lapse imaging. Treatment of GBP-HH significantly increased the stability of vinculin-positive focal adhesion complexes, as indicated by increased time-span between de novo assembly of focal adhesion and the final disassembly during imaging. Taken together, these results implied that GBP-HH may slow down disassembly of focal adhesion complex and thereby suppressed cell motility. Molecular mechanism underlying these intracellular processes is currently under investigation.

Tung-Kung Wu completed his PhD degree in Biophysics from the Johns Hopkins University and postdoctoral studies in Chemistry Department from Stanford University. He is appointed professor in the Department of Biological Science and Technology at the National Chiao Tung University. His research interests include interdisciplinary fields of bioorganic chemistry, chemical biology, and medicinal chemistry. His current research interests mainly focus on the structure-reactivity relationships of glycoenzymes and mode of action of steryl glycosides as drug candidates.

Steryl glycosides consist of a sterol/steroid aglycone and one or more sugar moieties linked through an ether or ester bond. Diverse biological activities, including anti-inflammatory, anti-fungal, anti-microbial, anti-parasitic, and anti-tumor activities, and properties such as cell membrane modification, regulation of host defenses against pathogens, lipid metabolism, and developmental events, have been attributed to this family of natural products. Development of structural diverse steryl glycosides may hold potential to identify candidates for new drug development and pharmaceutical applications. In this study, we functionally express different glycosyltransferases for in vitro synthesis of steryl glycosides with - or -configuration specificity. The synthesized steryl glycosides were investigated for their anticancer activity. Interestingly, several steryl glycosides show dose-dependent depression of cell viability and enhanced drug effectiveness on MCF-7 breast cancer cells. The structure-reactivity relationships of these glycosyltransferases on substrate‒donor specificities and mode of action of steryl glycosides on anticancer activity will be discussed.

Eduardo Osinaga is Medical Doctor and completed his PhD at the Technological University of Compiègne, France. He is the director of the Laboratory of Glycobiology and Tumor Immunology at Pasteur Institut, Montevideo, and is the Head of the Department of Immunobiology, Faculty of Medicine, Montevideo, Uruguay. His major research interests is the glyco-immunology of cancer. He has published more than 75 papers in reputed journals.

Lung cancer is currently the leading cause of cancer-related death worldwide, accounting for approximately a third of all cancer diagnoses and related-deaths. Non–small cell lung cancer (NSCLC) represents nearly 80% of lung tumors; the two most common NSCLC histological types are squamous cell carcinoma (SCC) and adenocarcinoma (ADCA). Several diagnosis procedures detecting altered glycosylation have been developed and incorporated as assistant procedures in clinical oncology. The synthesis of mucin-type O-glycans is started in a reaction catalyzed by UDP-N-acetyl-D-galactosamine:polypeptide N-acetylgalactosaminyltransferases (GalNAc-Ts). This is a complex family of enzymes of at least 20 members in humans. We previously found that GALNT13, the gene encoding GalNAc-T13 isoenzyme, is a strong predictor of poor clinical outcome in neuroblastoma patients (Berois et al., Clin Chem. 2006; 52:1701-12). In the present study, we evaluated GalNAc-T13 expression in human NSCLC. We produced a monoclonal antibody (MAb T13.5) that was used to assess the expression profile of the GalNAc-T13 protein in a well-defined population of 443 surgically resected NSCLC patients with 7 years of follow-up. We found that ADCAs expressed higher levels of the enzyme than SCCs. GalNAc-T13 expression correlated significantly with worse overall survival in ADCA patients treated with neoadjuvant chemotherapy. These data suggest that GalNAc-T13 could be a novel marker associated to chemoresistance in lung adenocarcinomas.