Glycans in Perspectives Mardi 21 Mars 2017 À Partir De 10H00 Salle Des Séminaires
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Mini-symposium de l’IPBS Glycans in Perspectives Mardi 21 Mars 2017 à partir de 10h00 Salle des séminaires 10h00-10h40 Dr. Weston Struwe Oxford Glycobiology Institute, Department of Biochemistry, University of Oxford, United Kingdom “Understanding Broadly Neutralizing Antibody and Anti-Viral Lectin Binding to HIV through Glycomics Analysis of Envelope Spikes” 10h40-11h20 Dr. Salomé S. Pinho Glycobiology in Cancer group, Institute for Research and Innovation in Health (i3S), University of Porto, Portugal “Glycosylation in Cancer: mechanisms and clinical implications” 11h20-12h00 Pr. Franck Fieschi Institut de Biologie Structurale, Université Grenoble Alpes, Grenoble “Sugar-mimetics as inhibitors of host- pathogen interaction : the case study of C- type Lectin targeted compounds” Contact : [email protected] & [email protected] Campus CNRS, 205 route de Narbonne - TOULOUSE Dr. Weston Struwe Oxford Glycobiology Institute, Department of Biochemistry, University of Oxford, United Kingdom Understanding Broadly Neutralizing Antibody and Anti-Viral Lectin Binding to HIV through Glycomics Analysis of Envelope Spikes Our research focuses on understanding HIV glycosylation and how antibodies and lectins recognize carbohydrate epitopes on the envelope glycoprotein (Env) complex. Env is trimer of gp120/gp41 dimer subunits and is 50% carbohydrate by mass. Crucially broadly neutralizing antibodies (bnAbs), which are elicited in up to 30% of infected individuals, target Env. The recent development of soluble Env mimics holds tremendous promise for an HIV vaccine and can provide passive protection in animal models. The abundance of Env glycans effectively shields the underlying peptides from innate immune responses but paradoxically many bnAbs target the viral spike via contact with conserved N-glycans. Lectins that have anti-viral properties have recently emerged as an alternative treatment to prevent HIV transmission, acting as microbiocides applied topically or even produced in situ by live engineered commensal microbes. Both bnAbs and microbiocide lectins target the HIV “glycan shield” yet the precise mechanisms by which they target Env is unclear. Importantly resistance to lectins among some HIV strains highlights the need to unpin these mechanisms. Our approach utilizes native and ion mobility mass spectrometry techniques as well as HPLC, NMR and molecular dynamics to uncovering the detailed molecular processes of these protein-carbohydrate interactions. Through site-specific, glycan and intact analysis of Env, we aim shed light on bnAb/lectin targeting of viral spikes in HIV vaccine and microbiocide development. References Struwe W, Stuckmann A, Behrens A, Pagel K, Crispin M. Global N-glycan Site Occupancy of HIV-1 gp120 by Metabolic Engineering and High-Resolution Intact Mass Spectrometry. ACS Chemical Biology. 2017 Behrens A, Harvey D, Milne E, Cupo A, Kumar A, Zitzmann N, Struwe W, Moore J, Crispin M. Molecular Architecture of the Cleavage-Dependent Mannose Patch of an HIV-1 Envelope Glycoprotein. Journal of Virology. 2017 Hofmann J, Stuckmann A, Crispin M, Harvey D, Pagel K, Struwe W. Identification of Lewis and Blood Group Carbohydrate Epitopes by Ion Mobility-Tandem-Mass Spectrometry Fingerprinting. Analytical Chemistry. 2017 Hopper J, Grant O, Ambrose S, Krumm S, Allison T, Tully M, Prichard L, Ozorowski G, Ward A, Crispin M, Doores K, Woods R, Benesch J, Robinson C, Struwe W. The Tetrameric Plant Lectin BanLec Neutralizes HIV through Bidentate Binding to Specific Viral Glycans. In Review Struwe W, Baldauf C, Hofmann J. Rudd P, Pagel K. Ion Mobility Separation of Deprotonated Oligosaccharide Isomers - Evidence for Gas-Phase Charge Migration. Chemical Communications. 2016 52(83): 12353-12356 Campus CNRS, 205 route de Narbonne - TOULOUSE Dr. Salomé S. Pinho Glycobiology in Cancer group Institute for Research and Innovation in Health (i3S), University of Porto, Portugal Glycosylation in Cancer: mechanisms and clinical implications Despite recent progress in understanding the cancer genome, there is still a relative delay in understanding the full aspects of the glycome and glycoproteome of cancer. Glycobiology has been instrumental in relevant discoveries in various biological and medical fields, and has contributed to the deciphering of several human diseases. Glycans are involved in fundamental molecular and cell biology processes occurring in cancer, such as cell signalling and communication, tumour cell dissociation and invasion, cell–matrix interactions, tumour angiogenesis, immune modulation and metastasis formation. The roles of glycans in cancer have been highlighted by the fact that alterations in glycosylation regulate the development and progression of cancer, serving as important biomarkers and providing a set of specific targets for therapeutic intervention. This Review discusses the role of glycans in fundamental mechanisms controlling cancer development and progression, and their applications in oncology. References Carvalho S, Catarino TA, Dias AM, Kato M, Almeida A, Hessling B, Figueiredo J, Gärtner F, Sanches JM, Ruppert T, Miyoshi E, Pierce M, Carneiro F, Kolarich D, Seruca R, Yamaguchi Y, Taniguchi N, Reis CA, Pinho SS. Preventing E- cadherin aberrant N-glycosylation at Asn-554 improves its critical function in gastric cancer. Oncogene. 2016 Mar 31;35(13):1619-31. doi: 10.1038/onc.2015.225. Carvalho S, Oliveira T, Bartels MF, Miyoshi E, Pierce M, Taniguchi N, Carneiro F, Seruca R, Reis CA, Strahl S, Pinho SS. O-mannosylation and N-glycosylation: two coordinated mechanisms regulating the tumour suppressor functions of E-cadherin in cancer. Oncotarget. 2016 Oct 4;7(40):65231-65246. doi: 10.18632/oncotarget.11245. Pinho SS, Reis CA. Glycosylation in cancer: mechanisms and clinical implications. Nat Rev Cancer. 2015 Sep;15(9):540-55. doi: 10.1038/nrc3982. Review Campus CNRS, 205 route de Narbonne - TOULOUSE Pr. Franck Fieschi Institut de Biologie Structurale, Université Grenoble Alpes, Grenoble Sugar-mimetics as inhibitors of host- pathogen interaction : the case study of C- type Lectin targeted compounds Our groups have been working now for years on C-type lectin receptors of dendritic cells and more particularly on DC-SIGN and langerin receptors. DC-SIGN has been implicated in numerous infection mechanisms associated to viruses (HIV, Ebola, HCV, etc…), bacteria, fungi and parasites. In genital mucosa, different fates are described for HIV according to the sub-type of dendritic cells (DCs) involved in its recognition. It notably depends on the C- type lectin receptor, langerin or DC-SIGN, involved in gp120 interaction. Langerin blocks HIV transmission by its internalization in specific organelles of Langerhans cells. On the contrary DC-SIGN enhances HIV trans-infection of T lymphocytes. Thus, approaches aiming to inhibit DC-SIGN, without blocking langerin, represent attractive strategies. In a more general context, C-type lectin receptors are involved in immune regulation and may activate an immune response or induce a tolerogenic response as a function of the receptors targeted and of the ligands encountered. There is great need to develop some specific ligands for the various receptors of this family in order to tailor the immune response as a function of the therapeutic needs. Development of such glycomimetics, rather than metabolically unstable carbohydrates, faces many bottlenecks directly link to the receptor family (with largely open active site with low selectivity), and also of the nature of natural carbohydrate that present low affinity to these receptors (at the mM level). Using DC-SIGN and Langerin as case study, approaches aiming to define new compounds with affinity reaching up to the nM level and increased specificity for one of the two receptor will be presented. Additional tools and development related to other C-type lectins, that are under progress, might be also discussed if time allows. References Thépaut, M., Guzzi, C., Sutkeviciute, I., Sattin, S., Ribeiro-Viana, R., Varga, N., Chabrol, E., Rojo, J., Bernardi, A., Angulo, J., Nieto, P. M., and Fieschi, F. Structure of a Glycomimetic Ligand in the Carbohydrate Recognition Domain of C-type Lectin DC-SIGN. Structural Requirements for Selectivity and Ligand Design. J Am Chem Soc 135, 2518–2529. 2013 Varga, N., Sutkeviciute, I., Ribeiro-Viana, R., Berzi, A., Ramdasi, R., Daghetti, A., Vettoretti, G., Amara, A., Clerici, M., Rojo, J., Fieschi, F., and Bernardi, A. A multivalent inhibitor of the DC-SIGN dependent uptake of HIV-1 and Dengue virus. Biomaterials 35, 4175–4184. 2014 Ordanini, S., Varga, N., Porkolab, V., Thépaut, M., Belvisi, L., Bertaglia, A., Palmioli, A., Berzi, A., Trabattoni, D., Clerici, M., Fieschi, F., and Bernardi, A. Designing nanomolar antagonists of DC-SIGN- mediated HIV infection: ligand presentation using molecular rods. Chem Commun (Camb) 51, 3816– 3819. 2015 Campus CNRS, 205 route de Narbonne - TOULOUSE .