Shotgun functional glycomics of heparan sulphate saccharides: generating diverse libraries to decode biological selectivity
Lead Research Organisation:
University of Liverpool
Department Name: Sch of Biological Sciences
Abstract
Complex sugars (glycans) are a highly diverse family of molecules with a broad range of functions in biological processes including cell recognition, adhesion, cell-cell communication and signalling. The study of the glycome - the entire set of glycans expressed by particular cells or tissues - is an emerging field which aims to understand how glycan functions underpin the complexity of human biology, and their involvement in disease processes. The selective interaction of proteins with glycans is one of the keys to their biological functions. A number of new technologies have emerged to support the development of 'glycomics' studies - large-scale studies of glycan structural diversity and the selective interactions of glycans with their matching protein partners which underpins their functions. However, these approaches do not directly reveal functional properties, especially for some complex classes of glycans like the heparan sulphate (HS) family of sulphated glycans which are responsible for regulation of a wide range of biological processes including growth factor signalling, enzyme activity and cell adhesion. We now need to have the tools to ask how specific structures control specific proteins to control biological systems. In this project we propose to develop a new approach to address this question by generating a large library of novel HS glycans with a wide range of structures, and screening them in biological assays, followed by analysis of selected structures. This will permit us to evaluate the structure-activity relationships of HS at higher throughput for the first time. This 'shotgun' library approach will provide a powerful and generic new tool for decoding the function of the HS glycome by allowing specific glycan structures to be matched to specific biological functions. In the future this strategy could provide new information that could be translated into applications in biotechnology and drug development.
Technical Summary
Decoding post-translational modifications of proteins, including the glycome, is a critical facet of the post-genome era, since they modify the functional proteome. Strategies have emerged to support 'glycomics' (large-scale) studies of glycan structure diversity and their selective interactions with cognate proteins. However to date these approaches have lacked the ability to directly generate functional data, particularly in the case of the complex heparan sulphate (HS) glycans. There is growing evidence that functional specificity exists in HS-dependent control of cognate protein activities, but technologies to truly address this question have been a bottleneck. The aim of this project is to develop a 'shotgun' functional glycomics approach for exploring the structure-activity relationships of HS. Generating a large random (unbiased) library of novel and diverse saccharides within HS chemical space, and screening them in test-bed bioassays, will permit detailed evaluation of biological specificity at higher throughput for the first time. We will: 1. Extend the chemical space of natural HS saccharide libraries covered by our existing pilot library (using a wider range of starting materials and saccharide generation methods). 2. Enhance the diversity of the HS library using enzymic approaches (recombinant sulfotransferases in concert with novel natural, semi-synthetic and synthetic substrates). 3. Screen the HS libraries to identify hits in test-bed bioassays (including FGF growth factor and Slit/Robo signalling). 4. Initiate structural analysis of HS library components and screening hits (to confirm structural diversity and identify novel structure-activity relationships, using state-of-the-art MS methods). This project will generate unique resources and a powerful new strategy for decoding the functions of the HS glycome. Such data will provide high value functional information on structure-activity relationships for the HS family.
Planned Impact
Expected Beneficiaries: This project is expected to have wide impact in many areas of biomedicine and biotechnology related to medicine in particular (eg. applications in diagnostics, drug discovery and regenerative medicine). This is due firstly to the relevance of HS biology to many disease processes (eg. cancer, inflammation, neurodegeneration, wound repair) and also fundamental biological processes that are critical for stem cell control and tissue engineering. New methods for determining specific HS targets will open up opportunities for breakthroughs in identification of novel information on HS specifity of biological action which could underpin commercial exploitation. More widely, the glycomics strategies will be of broad interest in view of the wide application potential of glycans in general. Better understanding of HS biology is also relevant to societal impacts. For example, Prof. Turnbull has met with families involved in the UK Hereditary Multiple Exostoses (HME) Support group (www.hmesg.org.uk). HME is caused by genetic deficiencies in HS biosynthesis that result in a multifactorial clinical problems including growth deficiency, bone tumours and premature death. The HME group are interested in promoting better understanding of the disease, current research into its causes and symptoms, and potential new treatments. Communications and Engagement: Commercial: The Turnbull lab has a number of active collaborations with Industry both in the UK and overseas, including IRL Ltd (a partner on this project), and research collaborations with SpheriTech Ltd (Runcorn; BBSRC CASE) and Summit (Dextra) Ltd (Reading). All these projects will benefit from the project and we will actively develop these partnerships as described in the Impact plan statement. Prof. Turnbull is also actively involved in discussions with the NorthWest Development Agency regarding the establishment of a Centre of Excellence in Glycosciences, aimed at networking of the high level of academic expertise in this field in the northwest of England with commercial partners. Societal: The UK Hereditary Multiple Exostoses (HME) Support group has a Liverpool group contact, Tina Read, who is developing web-based resources and information for families with children affected by this disease. The Turnbull group plan to assist them with information on the molecular basis of the disease and how future research might help with new disease treatments, and hosting of a meeting of the national organisation in Liverpool in 2010. General dissemination: We will actively seek to disseminate information about our research efforts to both industry and the general public, through websites (Liverpool Centre for Glycobiology, and University Business Gateway); press releases and opportunities for public speaking. Collaboration: The principal commercial partnership within the proposed project is with IRL Ltd (Wellington, NZ). We have an existing research agreement initiated in 2009 with IRL Ltd on identification of synthetic targets with potential commercial applications in Alzheimers disease and cancer. IRL will have an active interest in outputs regarding new targets and enzymic modification of their synthetic saccharides. Exploitation and Application: There is considerable potential for commercial exploitation of outputs from this project, for example in drug development, stem cell exploitation and tissue engineering. The tools and strategy development aspect of the project may also yield new intellectual property of commercial potential. We will actively and regularly monitor our research output and potential publications with ULive Ltd (the University of Liverpool IP and tech transfer company), protect by patenting, and exploit via out-licencing or development of a spin-out company. For full details see the Impact Plan appended
Organisations
- University of Liverpool (Lead Research Organisation)
- Chinese Academy of Sciences (Collaboration)
- University of Copenhagen (Collaboration)
- UNIVERSITY OF LIVERPOOL (Collaboration)
- IMPERIAL COLLEGE LONDON (Collaboration)
- Crown Research Institutes (Project Partner)
- University of North Carolina System (Project Partner)
Publications
Bromfield S
(2014)
Nanoscale self-assembled multivalent (SAMul) heparin binders in highly competitive, biologically relevant, aqueous media
in Chemical Science
Gray CJ
(2017)
Label-Free Discovery Array Platform for the Characterization of Glycan Binding Proteins and Glycoproteins.
in Analytical chemistry
Guimond, SE
Novel ex vivo transgenic mouse brain tissue assay for screening BACE1 inhibitors
in Nature Methods
Kim SH
(2011)
Extracellular matrix and cell signalling: the dynamic cooperation of integrin, proteoglycan and growth factor receptor.
in The Journal of endocrinology
Leary JA
(2015)
Composition, sequencing and ion mobility mass spectrometry of heparan sulfate-like octasaccharide isomers differing in glucuronic and iduronic acid content.
in European journal of mass spectrometry (Chichester, England)
Lettow M
(2020)
Cryogenic Infrared Spectroscopy Reveals Structural Modularity in the Vibrational Fingerprints of Heparan Sulfate Diastereomers.
in Analytical chemistry
Lord M
(2020)
Better growth-factor binding aids tissue repair.
in Nature biomedical engineering
Miller RL
(2020)
Shotgun ion mobility mass spectrometry sequencing of heparan sulfate saccharides.
in Nature communications
Miller RL
(2016)
Heparin Isomeric Oligosaccharide Separation Using Volatile Salt Strong Anion Exchange Chromatography.
in Analytical chemistry
Miller RL
(2016)
Enrichment of Two Isomeric Heparin Oligosaccharides Exhibiting Different Affinities toward Monocyte Chemoattractant Protein-1.
in Analytical chemistry
Miller RL
(2017)
Versatile Separation and Analysis of Heparan Sulfate Oligosaccharides Using Graphitized Carbon Liquid Chromatography and Electrospray Mass Spectrometry.
in Analytical chemistry
O'Neill P
(2017)
Sulfatase-mediated manipulation of the astrocyte-Schwann cell interface.
in Glia
Puvirajesinghe TM
(2012)
Array-based functional screening of heparin glycans.
in Chemistry & biology
Schwörer R
(2013)
Synthesis of a targeted library of heparan sulfate hexa- to dodecasaccharides as inhibitors of ß-secretase: potential therapeutics for Alzheimer's disease.
in Chemistry (Weinheim an der Bergstrasse, Germany)
Taylor SL
(2019)
By-Products of Heparin Production Provide a Diverse Source of Heparin-like and Heparan Sulfate Glycosaminoglycans.
in Scientific reports
Thompson SM
(2011)
Structure and epitope distribution of heparan sulfate is disrupted in experimental lung hypoplasia: a glycobiological epigenetic cause for malformation?
in BMC developmental biology
Turnbull JE
(2011)
Chemistry. Getting the farm out of pharma for heparin production.
in Science (New York, N.Y.)
Turnbull, JE
Identification of Orally Available Synthetic Heparinoids with Potent BACE1 Inhibitory Properties
in Nature Biotechnology
Zubkova, O
Heparan sulphate glycodendrimers with potent bioactivities: tools for chemical biology and therapeutic development.
in Angewandte Chemie
Description | Developed novel approaches for making and screening HS saccharide libraries |
Exploitation Route | Further research and application |
Sectors | Healthcare Pharmaceuticals and Medical Biotechnology |
Title | Engineered heparins |
Description | Libraries of selectively chemically modified heparins for applications in heparin-based drug discovery. |
Type Of Material | Technology assay or reagent |
Year Produced | 2006 |
Provided To Others? | Yes |
Impact | Exploited for a number of publications and to develop drug leads for AD, malaria, ant-cancer therapeutics and spinal repair therapies. |
Title | Heparan sulphate saccharide libraries |
Description | Development of technologies for and production of complex libraries of HS saccharides |
Type Of Material | Biological samples |
Year Produced | 2006 |
Provided To Others? | Yes |
Impact | Novel discoveries of structure-activity relationships for complex HS saccharides of relevance to biological and disease processes |
Description | Advanced mass spectrometry for glycomics |
Organisation | Imperial College London |
Department | Department of Life Sciences |
Country | United Kingdom |
Sector | Academic/University |
PI Contribution | Provision of compounds for analysis by MS |
Collaborator Contribution | Collaboration on advanced glycan analysis by mass spectrometry |
Impact | Network grants and publications |
Description | Collaboration with Chinese Academy of Sciences |
Organisation | Chinese Academy of Sciences |
Department | Institute of Material Medicine |
Country | China |
Sector | Academic/University |
PI Contribution | Partner in developing China-UK partnership. Expertise and contacts in polysaccharide research in China. |
Collaborator Contribution | Prof Kan Ding is leading efforts to develop this link through the network meetings. |
Impact | Ongoing partnership in development. |
Start Year | 2016 |
Description | Collaboration with Copenhagen University |
Organisation | University of Copenhagen |
Country | Denmark |
Sector | Academic/University |
PI Contribution | Developing collaboration on state of the art cell engineering and mass space sequencing technologies for glycosaminoglycans |
Collaborator Contribution | World class facilities at Centre for Glycomics |
Impact | Publications in Nature Methods and Nature Communications |
Start Year | 2019 |
Description | Glycomics strategies for heparan sulphates |
Organisation | University of Liverpool |
Department | School of Biological Sciences Liverpool |
Country | United Kingdom |
Sector | Academic/University |
PI Contribution | Expertise on structure and activity relationships and glycomics strategies |
Collaborator Contribution | Collaboration on molecular interactions and interactomics approaches. |
Impact | Publications, grants and ongoing research collaborations. |
Company Name | IntelliHep |
Description | IntelliHep develops a precision therapeutics for diseases such as Alzheimers and cancers. |
Year Established | 2002 |
Impact | Early stage development of hit compounds as beta-secretase inhibitors for treating the underlying cause of Alzheimers disease. Option to licence IP from Liverpool University on an anticancer metastatic drug. Employment of research staff and regular funded projects sub-contracted into University labs. Collaborating with China partners on commercial applications of heparin by-products. |
Website | http://www.intellihep.com |
Description | Hosting visit to Liverpool University by local ARUK funding supporters |
Form Of Engagement Activity | Participation in an open day or visit at my research institution |
Part Of Official Scheme? | Yes |
Type Of Presentation | Keynote/Invited Speaker |
Geographic Reach | Regional |
Primary Audience | Supporters |
Results and Impact | Contact was invaluable in informing supporters directly about research activities supported in Liverpool by their fund raising efforts Funding supporters were able to gain insights into new potential treatments for AD, and how their funding is used for research. |
Year(s) Of Engagement Activity | 2013 |
Description | Media releases on research findings |
Form Of Engagement Activity | A press release, press conference or response to a media enquiry/interview |
Part Of Official Scheme? | No |
Geographic Reach | National |
Primary Audience | Public/other audiences |
Results and Impact | University press releases on the novel Alzheimer drug hits we discovered, along with developments in synthetic chemistry of our sugars, and applications in nerve repair and cancer therapeutics International press coverage, radio interviews (Radio City Liverpool), and TV news (BBC NW Tonight). |
Year(s) Of Engagement Activity | 2006,2007,2008,2012,2013,2014 |
Description | Outreach to Business Community |
Form Of Engagement Activity | A magazine, newsletter or online publication |
Part Of Official Scheme? | No |
Geographic Reach | National |
Primary Audience | Health professionals |
Results and Impact | Article in magazine Research Intelligence by UoL Business Gateway,(see website link "Sweet Smell of Success" http://www.liv.ac.uk/researchintelligence/issue30/alzheimers.html) Dissemination of information to business community |
Year(s) Of Engagement Activity | 2007 |
URL | http://www.liv.ac.uk/researchintelligence/issue30/alzheimers.html |
Description | Public Lecture to Garston Rotary Club |
Form Of Engagement Activity | A talk or presentation |
Part Of Official Scheme? | No |
Geographic Reach | Local |
Primary Audience | Public/other audiences |
Results and Impact | Lecture to Garston Rotary Club Contributing the Public Understanding of Science |
Year(s) Of Engagement Activity | 2008 |
Description | Public lecture to Liverpool Soroptimists Club |
Form Of Engagement Activity | A talk or presentation |
Part Of Official Scheme? | Yes |
Geographic Reach | Local |
Primary Audience | Public/other audiences |
Results and Impact | Talk sparked many questions and discussion of AD and potential new drugs and diagnostics Contribution to Public Understanding of Science |
Year(s) Of Engagement Activity | 2014 |
Description | RCUK/BBSRC event: Lifelong Health: Bioscience of Ageing (at Westminster) |
Form Of Engagement Activity | A talk or presentation |
Part Of Official Scheme? | No |
Geographic Reach | National |
Primary Audience | Policymakers/politicians |
Results and Impact | Poster highlighting discovery of new class of drugs for AD at the RCUK/BBSRC event: 2007 Lifelong Health: Bioscience of Ageing (at Westminster) Dissemination of information on research success to MPs and policymakers |
Year(s) Of Engagement Activity | 2007 |
Description | Supporting ARUK fund raising publicity with major London donor |
Form Of Engagement Activity | Participation in an activity, workshop or similar |
Part Of Official Scheme? | Yes |
Type Of Presentation | Keynote/Invited Speaker |
Geographic Reach | National |
Primary Audience | Supporters |
Results and Impact | Supporters engaged directly with researcher undertaking research activity. As above |
Year(s) Of Engagement Activity | 2013 |
Description | Visit to Coleg Cambria, Wrexham. Meet the Scientist & Careers Talk. February 2019. |
Form Of Engagement Activity | A talk or presentation |
Part Of Official Scheme? | No |
Geographic Reach | Local |
Primary Audience | Schools |
Results and Impact | Visit to Coleg Cambria, Wrexham. Meet the Scientist & Careers Talk. February 2019. |
Year(s) Of Engagement Activity | 2019 |