Microarray bioassays for functional glycome screening

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 the key to their biological functions. A number of new technologies have emerged to support such interaction studies, chief among these being microarray techniques ('glycoarrays'), which allow large numbers of individual glycans to be immobilised on surfaces for examination of their selective binding to protein partners. However, whilst allowing rapid large-scale screening, glycoarrays generate simple interaction data that do not directly reveal functional properties. In this project we propose to develop a novel glycoarray methodology in which live cell responses to large numbers of glycans arrayed on specially modified microscope slides are measured quantitatively. Responses such as cell growth or differentiation will be analysed by fluorescence measurements, using either antibodies or induced expression of fluorescent proteins by the cells themselves. This approach will provide a powerful and generic new tool for decoding the function of the glycome by allowing specific glycan structures to be matched to specific biological functions.

Technical Summary

Glycans are a highly diverse class of molecules with myriad functions in biological processes including cell recognition, adhesion and communication. 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 the functions of glycans underpins biological regulation and complexity. Decoding the glycome will be a critical facet of the post-genome era, in partnership with other post-translational modifications which modify with the functional proteome. A number of new technologies have emerged to support studies of the selective interactions of glycans with their cognate protein partners which underpins their functions. Chief among these are recent microarray techniques ('glycoarrays') which allow large numbers of individual glycans to be immobilised on surfaces for screening their selective binding to protein partners. However, whilst allowing rapid large-scale screening, glycoarrays generate simple interaction data that does not directly reveal functional properties. Here we propose to develop a novel glycoarray methodology in which live cell responses to large numbers of glycans arrayed as slide-based microarrays are measured quantitatively. Such data will provide high value functional information on glycan specificity. We will: 1. Develop a slide-based microarray methodology for large-scale analysis of live cell responses to immobilized glycans. 2. Test the application of this methodology to quantitative screening of cell responses to heparan sulphate saccharides (eg. signalling pathway activation; differentiation). 3. Evaluate various potential detection methods for cell responses (eg. immunofluorescence; induction of endogenous fluorescent proteins). 4. Explore the generic application of the methodology using a different class of glycans (sialyl N-glycans). This project will provide a powerful and generic new tool for decoding the functions of the glycome.

Publications

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Puvirajesinghe TM (2012) Array-based functional screening of heparin glycans. in Chemistry & biology

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Turnbull JE (2010) Glycomics profiling of heparan sulfate structure and activity. in Methods in enzymology

 
Description DEveloped a novel microarray platform for measuring live cell responses to immobilised sugars
Exploitation Route Further research
Sectors Healthcare,Manufacturing, including Industrial Biotechology,Pharmaceuticals and Medical Biotechnology

 
Description Technology still under development
First Year Of Impact 2012
Sector Pharmaceuticals and Medical Biotechnology
 
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
 
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