dr. juan echevarria - wordpress.com | annual report 4 opportunities for glyconanomaterials in...

2 Glycotechnology | Annual Report

The Glycotechnology Group at CIC biomaGUNE is engaged in both basic and applied research in Glycoscience.

Glycoscience is still a relatively understudied area but has a strong potential to provide groundbreaking solutions to major societal challenges in personalized medicine, pharmaceuticals, food and biomaterials.

In our research we focus specifically on Biomedical Glycoscience, investigating and exploiting structure and function of carbohydrates in the context of human biology and disease.

We do this by developing specific tools for glycan analysis and glycan-based intervention and by closely collaborating with a growing number of internationally recognized researchers.

The following pages illustrate the active major research lines in 2016 and our efforts in translating our results from basic science into commercially viable products.

Our main areas of research are:

• Carbohydrate Synthesis

• Glycan Microarrays

• Lectin Analysis

• Glycan Analysis

• Chemical Immunology

3

Dr. Juan Echevarria Postdoctoral Researcher

(until Oct.16)

Dr. Alvaro Hernández Postdoctoral Researcher

Dr. Sonia Serna Postdoctoral Researcher

Raquel Pazos Research Technician

Elisabete Elosegui Research Technician

Begoña Echeverría Research Technician

Julie Pham IMMUNOSHAPE

PhD Student

Dr. Niels-Christian Reichardt Principal Investigator

Dr. Manuel Martín Lomas CIC biomaGUNE Founding Director

Anna Cioce IMMUNOSHAPE

PhD Student

Bárbara Kuhn PhD Student

Dr. Cristina Díez IMMUNOSHAPE Project Manager

Emma Sery Joint PhD Student with University of Liverpool

Charles Williams Joint PhD Student with CIC bioGUNE

Glycotechnology | Annual Report

Dr. Nerea Ruiz Postdoctoral Researcher

María Alcalá Research Technician

(substitution from Dec.16)

Dr. Olatz Villate IMMUNOSHAPE Project Manager

(substitution until May 16)

Katarzyna Brzezicka PhD Student (until Jan.16)


Opportunities for Glyconanomaterials in Personalized Medicine

Reichardt, N.-C.; Martin-Lomas, M. ; Penades, S.

Chem. Commun., 2016, 52 (92), 13430 – 13439 | DOI: 10.1039/C6CC04445J

In this feature article we discuss the particular relevance of glycans as components or targets of functionalized nanoparticles (NPs) for potential applications in personalized medicine but we will not enter into descriptions for their preparation. For a more general view covering the preparation and applications of glyconanomaterials the reader is referred to a number of recent reviews. The combination of glyco and nanotechnology is already providing promising new tools for more personalized solutions to diagnostics and therapy. Current applications relevant to personalized medicine include drug targeting, localized radiation therapy, imaging of glycan expression of cancer cells, point of care diagnostics, cancer vaccines, photodynamic therapy, biosensors, and glycoproteomics.

Influence of Core β-1,2-xylosylation on Glycoprotein Recognition by Murine C-type Lectin Receptors and its Impact on Dendritic Cell Targeting

Brzezicka K.; Vogel U., Serna S., Johannssen T., Lepenies B., Reichardt N.C.

ACS Chem. Biol. 2016, 11 (8), 2347–2356

Targeting antigens to dendritic cell subsets is a promising strategy to enhance the efficacy of vaccines. C-type lectin receptors (CLRs) expressed by dendritic cells are particularly attractive candidates since CLR engagement may promote cell uptake and may further stimulate antigen presentation and subsequent T cell activation. While most previous approaches have involved antibody-mediated CLR-targeting, glycan-based CLR targeting has become more and more attractive in recent years. In the present study, we show that small structural glycan modifications may markedly influence CLR recognition, dendritic cell targeting, and subsequent T cell activation. A biantennary N-glycan (G0) and its analogous O-2 core xylosylated Nglycan (XG0) were synthesized, covalently conjugated to the model antigen ovalbumin, and analyzed for binding to a set of murine CLR-Fc fusion proteins using lectin microarray. To evaluate whether the differential binding of G0 and XG0 to CLRs impacted dendritic cell targeting, uptake studies using murine dendritic cells were performed. Finally, effects of the ovalbumin glycoconjugates on T cell activation were measured in a dendritic cell/T cell cocultivation assay. Our results highlight the utility of glycan-based dendritic cell targeting and demonstrate that small structural differences may have a major impact on dendritic cell targeting efficacy.


Carbohydrate-Binding Non-Peptidic Pradimicins for the Treatment of Acute Sleeping Sickness in Murine Models

Castillo-Acosta V.M., Ruiz-Pérez L.M,. Etxebarria J,. Reichardt N.C., Navarro M., Igarashi Y., et al.

PLOS Pathogens | DOI:10.1371/journal.ppat.1005851

Current treatments available for African sleeping sickness or human African trypanosomiasis (HAT) are limited, with poor efficacy and unacceptable safety profiles. Here, we report a new approach to address treatment of this disease based on the use of compounds that bind to parasite surface glycans leading to rapid killing of trypanosomes. Pradimicin and its derivatives are non-peptidic carbohydrate-binding agents that adhere to the carbohydrate moiety of the parasite surface glycoproteins inducing parasite lysis in vitro. Notably, pradimicin S has good pharmaceutical properties and enables cure of an acute form of the disease in mice. By inducing resistance in vitro we have established that the composition of the sugars attached to the variant surface glycoproteins are critical to the mode of action of pradimicins and play an important role in infectivity. The compounds identified represent a novel approach to develop drugs to treat HAT.

Efficient monitoring of protein ubiquitylation levels using TUBEs-based microarrays

Serna, S., Xolalpa, W., Lang, V., Aillet, F., England, P., Reichardt, N., & Rodriguez, M. S.

FEBS letters, 2016, 590 (16), 2748–2756

Analyzing protein ubiquitylation changes during physiological or pathological processes is challenging due to its high reversibility and dynamic turnover of modified targets. We have developed a protein microarray to assess endogenous ubiquitylation levels from cell cultures, employing tandem ubiquitin binding entities (TUBEs) with three or four ubiquitin-associated (UBA) domains as capture probes. Adriamycin (ADR)-stimulated MCF7 cells were used to differentiate protein ubiquitylation levels between cells that are sensitive or resistant to ADR treatment. We show that TUBEs-based microarrays can be used for the analysis of cellular processes regulated by ubiquitylation and for the detection of pathologies with aberrant ubiquitylation levels.


Methods for the absolute quantification of N-glycan biomarkers

Etxebarria J. and Reichardt N.C.

Biochim. Biophys Acta, 2016, 1860 (8), 1676–1687

Many treatment options especially for cancer show a low efficacy for the majority of patients demanding improved biomarker panels for patient stratification. Changes in glycosylation are a hallmark of many cancers and inflammatory diseases and show great potential as clinical disease markers. The large intersubject variability in glycosylation due to hereditary and environmental factors can complicate rapid transfer of glycan markers into the clinical practice but also presents an opportunity for personalized medicine. This review discusses opportunities of glycan biomarkers in personalized medicine and reviews the methodology for N-glycan analysis with a specific focus on methods for absolute quantification. The entry into the clinical practice of glycan markers is delayed in large part due to a lack of adequate methodology for the precise and robust quantification of protein glycosylation. Only absolute glycan quantification can provide a complete picture of the disease related changes andwill provide themethod robustness required by clinical applications. Glycan biomarkers have a huge potential as diseasemarkers for personalizedmedicine. The use of stable isotope labeled glycans as internal standards and heavy-isotope labeling methods will provide the necessary method precision and robustness acceptable for clinical use.


Asparia - mAbsolute Glycan Standards for Diagnostics (Invited)

Niels Reichardt

Metrology of Carbohydrates, Brussels (Belgium)

18-19 February 2016

Synthesis and Biomedical Applications of Parasite glycans (Invited)

Niels Reichardt

XXVI Reunión Bienal de Química Orgánica, Punta Umbría (Spain)

14-17 June 2016

Absolute Quantification of N-glycans by Isotopic Dilution

Juan Echeverria, Begoña Echeverria, Alvaro Hernandez, Nerea Ruiz, Niels-Christian Reichardt

XXVIII ICS - International Carbohydrate Symposium, New Orleans (USA)

17-21 July 2016

Synthesis and Biomedical Applications of Parasite glycans Niels Reichardt


17-21 July 2016

Moving towards a common language (Member Roundtable)

Niels Reichardt

GEIVEC, San Sebastian (Spain)

October 2016

On-chip Synthesis of MALDI-TOF-MS detectable Glycan Microarrays

Giulio Goti, Anna Cioce, Sonia Serna, Anna Bernardi, Niels Reichardt

XLI International Summer School on Organic Synthesis “A. Corbella”, Gargnano (Italy)

12-17 June 2016


Assessing glycan-specific IgE profiles associated with S. mansoni infection and allergy in Uganda

G. Nkurunungi, A. van Diepen, J. Kabagenyi, R. Sanya, N. Reichardt, C. Hokke, A. Elliott, M. Yazdanbakhsh

Molecular and Cellular Biology of Helminth Parasites X, Hydra (Greece)

4-9 September 2016

Characterization of ubiquitylation defects associated with resistance to Adriamycin

Valérie Lang, Fabienne Aillet, Sonia Serna, Wendy Xolalpa, Patrick England, Niels Reichardt & Manuel S. Rodríguez

Proteostasis, Lisbon (Portugal)

2-5 November 2016

On-chip development of N-glycan mimetics for improving CLRs targeting

Anna Cioce, Sonia Serna, Álvaro Hernández, Giulio Goti, Anna Bernardi, Niels C. Reichardt


17-21 July 2016

On-chip development of N-glycan mimetics for improving CLRs targeting

Anna Cioce, Sonia Serna, Álvaro Hernández, Giulio Goti, Anna Bernardi, Niels C. Reichardt

11th Spanish – Italian Symposium on Organic Chemistry, San Sebastian (Spain)

13-15 July 2016

9

Katarzyna Brzezicka

Chemoenzymatic Synthesis and Immunological Studies of Xylosylated N-glycans

Director: Niels-Christian Reichardt University of Basque Country (UPV), San Sebastian / CIC biomaGUNE

June 7, 2016


Luca Unione

Sugars and Proteins: The Role of Dynamics in Molecular Interactions

Directors: Jesús J. Barbero – CIC bioGUNE, F. Javier Cañada Vicinay - CSIC-CIB Member of Jury: Niels Reichardt School of Pharmacy - Complutense University of Madrid (UCM)

October 18, 2016

Emma Christelle Sery

Chemical Biology Tools for Structure-Function Studies on Heparan Sulphates: Decoding Specificity in FGF Signalling

Directors: Niels-Christian Reichardt & Jerry Turnbull CIC biomaGUNE & University of Liverpool (UK)

International Joint Graduate School Programme

December 19, 2016


Silvia Achilli

Measuring the binding profiles of available recombinant human CLRs to glycans and glycomimetics in microarray screening assays

Home Institution: Institut de Biologie Structurale, IBS – Université Grenoble Alpes (France)

May 2016

Laura Medve

Preparation of glycomimetic arrays for screening with lectins

Home Institution: Università degli Studi di Milano (Italy)

June-July 2016

Joao Monteiro

Screening of chemically synthetized glycans for binding to a murine CLR-Fc fusion proteins library using lectin microarray

Home Institution: Research Center for Emerging Infections and Zoonoses – University of Veterinary Medicine Hannover (Germany)

October 2016

Valentin Cognet

Proof of concept of the binding ability between Midatech gold nanoparticles carrying carbohydrates and CIC biomaGUNE Lectins chips

Home Institution: Midatech Pharma, Bilbao (Spain)

November 2016

Julie-Thanh Pham

Expression and purification of C-type Lectin receptors DC-SIGN and DC-SIGN R

Host Institution: Institut de Biologie Structurale, IBS – Université Grenoble Alpes (France)

March 2016


Antoni Planas Specificity of chitin deacetylases: structural determinants defining the de-n-acetylation pattern

Institut Químic de Sarrià – Universitat Ramon Llull, Barcelona (Spain)

8 June 2016

Héctor Peinado Sellas

Exploring new frontiers, role of tumor-secreted exosomes in metastasis

Head of Microenvironment and Metastasis Group – CNIO, Madrid (Spain)

10 March 2016

Sander van Kasteren

Imaging Degradation

Bio-organic Synthesis, Chemistry Institute – School of Sciences, Leiden (The Netherlands)

13 April 2016

Bjoern Christensen

Alginate biomaterials , branched chitosan for gene delivery and β-1,3-glucans

Norwegian University of Science and Technology – NTNU, Troendheim (Norway)

11 May 2016

Juan Falcón Exosomes: metabolic nano-machines encoding complex signals

CIC bioGUNE, Bilbao (Spain)

18 May 2016


Some research results have been translated to the marketplace in 2016. A number of patented innovative technologies have resulted from the basic research developed in this group and are available upon request and via distributors.

1. Glycan standards, kits and enzymes are currently being commercialized via the Asparia Glycomics brand established in September 2016 by CIC biomaGUNE and private investors https://aspariaglycomics.com/

mAbsolute kit – reagents for absolute glycan quantification by MALDI-ToF-MS as well as custom made quantification software

2. Weathering steel LDI-MS sample plates – matrix-free small molecule analysis on conventional mass spectrometers

The company is specialized in the production and marketing of reagents, reference standards, kits and software for glycan analysis in clinical diagnosis and for quality control of biopharmaceuticals. The business initiative markets the leading technology developed by the Glycotechology laboratory led by Dr. Niels Reichardt to quantify glycans more accurately and faster than existing solutions on the market. The company is led by Dr. Juan Echevarria, a co-developer of the technology, who moved to Asparia Glyomics from the Glycotechnology laboratory. Current Asparia Products are:

CarboQuant –unique stable heavy-isotope labelled glycan standards as universal internal standards for N- and O-glycan analysis by mass spectrometry or other methods

Recombinant PNGase F, CarboClip

https://aspariaglycomics.com/


The establishment of the new spin-off company Asparia Glycomics in September 2016 and information about its products and services has been covered by several media.


CIBER-BBN

Glyco Thyroid Cancer

Intramural project

Etortek (Coordination)

IE14-385

Investigación estratégica y Desarrollo Tecnológico en Aplicaciones Biomédicas de Nanoglicotecnología y Espectrometría de Masas sobre Superficies para Diagnóstico Clínico y Seguridad Alimentaria

2014-2015

Spanish MINECO

GLYCOIMMUNOTECH

Grant No. CTQ2014-58779-R

Tools to study and exploit the role of glycans in adaptive and innate immunity

2015-2017

European Commission

IMMUNOSHAPE (Coordination)

H2020-MSCA-ITN-2014-ETN-642870

Development of Selective Carbohydrate Immunomodulators Targeting C-type Lectin Receptors on Antigen Presenting Cells

2015-2018

www.immunoshape.eu

http://www.immunoshape.eu/

15

Our multidisciplinary research is only possible in collaboration with a growing

number of international recognized researchers. Many current collaborations are

managed within the IMMUNOSHAPE project that runs until 2018.

• Prof. Bernd Lepenies, Infection Immunology. University of Veterinary Medicine

Hannover. Research Center for Emerging Infections and Zoonoses (RIZ), Germany

• Anna Bernardi, Chemistry Department. Università degli Studi di Milano, Italy

• Sabine Flitsch, Manchester Institute of Biotechnology (MIB). The University of

Manchester, UK

Other current collaborations include:

• Dr. Ron Hokke, Parasite Glycobiology Group. Department of Parasitology. University

Medical Center Leiden, Netherlands

• Prof. Jesús Jiménez-Barbero, Chemical Glycobiology Lab. CIC bioGUNE, Bilbao, Spain

• Dr. Jesús Angulo, School of Pharmacy. University of East Anglia, Norwich, UK

• Prof. Juan Falcón, Exosomes Lab. CIC bioGUNE , Bilbao, Spain

• Dr. Alberto de Leiva, Departament of Medicine and Director of Department of

Endocrinology and Nutrition. Hospital San Pau, Barcelona, Spain

• Dr. Emilio Pérez-Trallero, University Hospital Donostia, Spain

• Dr. Dolores Gonzales, Departament of Biochemistry and Molecular Pharmacology.

Instituto de Parasitología y Biomedicina-Lopez Neyra, Granada, Spain

• Dr. Héctor Peinado, Microenvironment and Metastasis Group. CNIO, Madrid, Spain

• Dr. Katie Doores, Infectious Diseases. Faculty of Life Sciences and Medicine. Kings

College London, UK

• Prof. Manfred Wuhrer, Center for Proteomics and Metabolomics. Leiden University

Medical Center, Netherlands

• Prof. Maurizio Prato, Carbon Bionanotechnology Group at CIC biomaGUNE and

Università degli Studi di Trieste, Italy



Farewell lunch for Olatz Villate at Basozabal Golf - May 2016

Christmas Lunch together with Maurizio Prato’s Group at CIC biomaGUNE - Dec 2016

dr. juan echevarria - wordpress.com | annual report 4 opportunities for glyconanomaterials in...

Documents