14-ERASynBio: Synthetic Glycobiology - new strategies to build and functionalise proto-cells and proto-tissues

Lead Research Organisation: University of Leeds
Department Name: Sch of Chemistry

Abstract

Great advances have been made in the development of proto-cells based on giant unilamellar vesicles (GUVs). However, one essential functional element of all living cells still to be incorporated into such systems is a glycocalyx. This coating of complex carbohydrates extends up to 100 nm from the cell membrane and provides an adhesive layer that mediates interactions between different cell types, viruses and signalling molecules. In most cases, these interactions involve specific carbohydrate-binding proteins (lectins) which may be either soluble or membrane-bound. For example, fertilisation is initiated by a specific carbohydrate on the surface of the egg adhering to a specific lectin on the head of the sperm. Protein-carbohydrate interactions also mediate the endocytosis of many bacteria, viruses and bacterial toxins which stick to specific glycolipids on the cell membrane. Protein-carbohydrate interactions thus present a general strategy for enabling cell adhesion and cell entry. In this project we will design and create a modular toolbox of synthetic glcocalyx components and engineered lectins that will be attached to lipid membranes to enable reversible proto-cell adhesion and incorporated into virus-like particles to mediate proto-cell entry. The methodology will be exemplified through the construction of proto-cells that contain "proto-organelles" and the assembly and remodelling of "proto-tissues" in which multiple types of proto-cells are brought together in a pre-defined fashion to create more complex systems.

Technical Summary

Great advances have been made in the development of proto-cells based on giant unilamellar vesicles (GUVs). However, one essential functional element of all living cells still to be incorporated into such systems is a glycocalyx. This coating of complex carbohydrates extends up to 100 nm from the cell membrane and provides an adhesive layer that mediates interactions between different cell types, viruses and signalling molecules. In most cases, these interactions involve specific carbohydrate-binding proteins (lectins) which may be either soluble or membrane-bound. For example, fertilisation is initiated by a specific carbohydrate on the surface of the egg adhering to a specific lectin on the head of the sperm. Protein-carbohydrate interactions also mediate the endocytosis of many bacteria, viruses and bacterial toxins which stick to specific glycolipids on the cell membrane. Protein-carbohydrate interactions thus present a general strategy for enabling cell adhesion and cell entry. In this project we will design and create a modular toolbox of synthetic glcocalyx components and engineered lectins that will be attached to lipid membranes to enable reversible proto-cell adhesion and incorporated into virus-like particles to mediate proto-cell entry. The methodology will be exemplified through the construction of proto-cells that contain "proto-organelles" and the assembly and remodelling of "proto-tissues" in which multiple types of proto-cells are brought together in a pre-defined fashion to create more complex systems.

Planned Impact

In the longer term, the neolectins, superlectins, virus-like particles and lectin-modified proto-cells will provide tools for delivering macromolecular drugs in a cell-specific fashion. At present most of the biopharmaceuticals on the market recognise cell surface targets, but the neolectins etc, will provide a mechanism for delivery into cells by endocytosis, and potentially intracellular trafficking to defined organelles. This advance will expand the range of medical targets accessible to biopharmaceuticals, while helping to reduce the high doses necessary for novel treatments that already act intracellularly, e.g., exon-skipping RNA based drugs. Applicant Turnbull already collaborates with GSK on macromolecular drug delivery using lectins. He also leads the targeted delivery theme of the University of Leeds Pharmaceutical and Biopharmaceutical Industry Hub which fosters translational collaborations between academic and industrial partners. Furthermore, the involvement of applicant Wiltschi from the Austrian Centre of Industrial Biotechnology (ACIB) will provide another key pathway to achieving impact from our research. The ACIB has 30 company partners and will function as a hub for the dissemination of results to potential industrial partners and the translational development of products involving neolectins and superlectins developed in SYNGLYCTIS.
Through this proposal we will contribute to the training of skilled individuals (including 8 early stage researchers) who will gain expertise in design, engineering and characterisation of protein and lipid assemblies for synthetic biology applications. The provision of skilled workers who are empowered to contribute to the development of both academic and industrial synthetic biology will have long term societal benefit for Europe. Their work in this project and through the rest of their careers will lead to new diagnostic methods and clinical treatments that will improve the health and quality of life for millions of Europeans.

Publications

10 25 50
 
Description Re-engineered carbohydrate-binding proteins and virus-like particles have been prepared and used as cross-linking agents to prepare artificial tissues. We have discovered that subtle changes in the architecture of the sugar-binding proteins can give rise to complex behaviours upon interactions with biological membranes such as membrane fusion.
Reengineered bacterial toxins have been used to deliver nanoparticles into motor neurones
Exploitation Route potential applications in drug delivery and tissue engineering
Sectors Healthcare,Pharmaceuticals and Medical Biotechnology

 
Description BBSRC DTP summer studentship
Amount £2,500 (GBP)
Organisation Biotechnology and Biological Sciences Research Council (BBSRC) 
Sector Public
Country United Kingdom
Start 07/2016 
End 08/2016
 
Description BBSRC Follow on Fund Proof of concept
Amount £15,999 (GBP)
Funding ID BB/P028152/1 
Organisation Biotechnology and Biological Sciences Research Council (BBSRC) 
Sector Public
Country United Kingdom
Start 03/2016 
End 07/2017
 
Description BBSRC Follow-on-Fund
Amount £199,902 (GBP)
Funding ID BB/R005540/1 
Organisation Biotechnology and Biological Sciences Research Council (BBSRC) 
Sector Public
Country United Kingdom
Start 05/2018 
End 10/2019
 
Description EPSRC Impact Acceleration Account
Amount £5,200 (GBP)
Organisation Engineering and Physical Sciences Research Council (EPSRC) 
Sector Academic/University
Country United Kingdom
Start 07/2016 
End 08/2016
 
Description H2020-MSCA-ITN-2018
Amount € 4,093,660 (EUR)
Funding ID 814029 
Organisation European Union 
Sector Public
Country European Union (EU)
Start 10/2018 
End 09/2022
 
Description IBCarb proof of concept funding
Amount £15,983 (GBP)
Funding ID BB/L013762/1 
Organisation Biotechnology and Biological Sciences Research Council (BBSRC) 
Sector Public
Country United Kingdom
Start 01/2018 
End 03/2018
 
Description Royal Society Theo Murphy International Scientific Meeting
Amount £10,000 (GBP)
Organisation The Royal Society 
Sector Academic/University
Country United Kingdom
Start 10/2018 
End 10/2018
 
Description White Rose BBSRC DTP
Amount £100,000 (GBP)
Organisation Biotechnology and Biological Sciences Research Council (BBSRC) 
Sector Public
Country United Kingdom
Start 10/2017 
End 09/2021
 
Description White Rose BBSRC DTP Industrial CASE
Amount £100,000 (GBP)
Organisation Biotechnology and Biological Sciences Research Council (BBSRC) 
Sector Public
Country United Kingdom
Start 10/2017 
End 09/2021
 
Description SynGlycTis 
Organisation Albert Ludwigs University of Freiburg
Country Germany 
Sector Hospitals 
PI Contribution Sharing of facilities, samples and expertise
Collaborator Contribution Providing detailed research methodologies and hosting researchers to perform specific experiments
Impact new methods for formation of prototissues and delivery of quantum dots into brain cells Funding for a Royal Society Theo Murphy International Meeting Funding for an EU Marie Sklodowska-Curie Initial Trainin network (synBIOcarb
Start Year 2014
 
Description SynGlycTis 
Organisation Austrian Centre of Industrial Biotechnology
Country Austria 
Sector Private 
PI Contribution Sharing of facilities, samples and expertise
Collaborator Contribution Providing detailed research methodologies and hosting researchers to perform specific experiments
Impact new methods for formation of prototissues and delivery of quantum dots into brain cells Funding for a Royal Society Theo Murphy International Meeting Funding for an EU Marie Sklodowska-Curie Initial Trainin network (synBIOcarb
Start Year 2014
 
Description SynGlycTis 
Organisation National Center for Scientific Research (Centre National de la Recherche Scientifique CNRS)
Department Centre National de la Recherche Scientifique Grenoble
Country France 
Sector Public 
PI Contribution Sharing of facilities, samples and expertise
Collaborator Contribution Providing detailed research methodologies and hosting researchers to perform specific experiments
Impact new methods for formation of prototissues and delivery of quantum dots into brain cells Funding for a Royal Society Theo Murphy International Meeting Funding for an EU Marie Sklodowska-Curie Initial Trainin network (synBIOcarb
Start Year 2014
 
Description SynGlycTis 
Organisation University of Copenhagen
Country Denmark 
Sector Academic/University 
PI Contribution Sharing of facilities, samples and expertise
Collaborator Contribution Providing detailed research methodologies and hosting researchers to perform specific experiments
Impact new methods for formation of prototissues and delivery of quantum dots into brain cells Funding for a Royal Society Theo Murphy International Meeting Funding for an EU Marie Sklodowska-Curie Initial Trainin network (synBIOcarb
Start Year 2014
 
Description SynGlycTis network 
Organisation Albert Ludwigs University of Freiburg
Department Centre for Biological Signalling Studies (BIOSS)
Country Germany 
Sector Academic/University 
PI Contribution We have provided expertise in protein modification and samples for testing
Collaborator Contribution Expertise in introducing non-canonical amino acids into proteins transferred to Leeds from AICB Samples from Leeds evaluated in giant unilamellar vesicle experiments at University of Freiburg
Impact Outputs ongoing. Collaboration brings together chemists, structural biologists, protein engineers, cell biologists.
Start Year 2014
 
Description SynGlycTis network 
Organisation Austrian Centre for Applied Biotechnology
Country Austria 
Sector Academic/University 
PI Contribution We have provided expertise in protein modification and samples for testing
Collaborator Contribution Expertise in introducing non-canonical amino acids into proteins transferred to Leeds from AICB Samples from Leeds evaluated in giant unilamellar vesicle experiments at University of Freiburg
Impact Outputs ongoing. Collaboration brings together chemists, structural biologists, protein engineers, cell biologists.
Start Year 2014
 
Description SynGlycTis network 
Organisation National Center for Scientific Research (Centre National de la Recherche Scientifique CNRS)
Country France 
Sector Public 
PI Contribution We have provided expertise in protein modification and samples for testing
Collaborator Contribution Expertise in introducing non-canonical amino acids into proteins transferred to Leeds from AICB Samples from Leeds evaluated in giant unilamellar vesicle experiments at University of Freiburg
Impact Outputs ongoing. Collaboration brings together chemists, structural biologists, protein engineers, cell biologists.
Start Year 2014
 
Description SynGlycTis network 
Organisation University of Copenhagen
Department Department of Chemistry
Country Denmark 
Sector Academic/University 
PI Contribution We have provided expertise in protein modification and samples for testing
Collaborator Contribution Expertise in introducing non-canonical amino acids into proteins transferred to Leeds from AICB Samples from Leeds evaluated in giant unilamellar vesicle experiments at University of Freiburg
Impact Outputs ongoing. Collaboration brings together chemists, structural biologists, protein engineers, cell biologists.
Start Year 2014
 
Description BioLeeds 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach Local
Primary Audience Public/other audiences
Results and Impact Collaboration with a group of artists to produce an exhibition for engaging the general public in glycoscience
Year(s) Of Engagement Activity 2015
 
Description Complex Life of Sugars 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? Yes
Geographic Reach National
Primary Audience Public/other audiences
Results and Impact stimulated the public's awareness of the importance of glycoscience

enthused several of my students to participate in further outreach eevnts
Year(s) Of Engagement Activity 2013,2014
 
Description Complex Life of Sugars @ Astbury Conversation 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Public/other audiences
Results and Impact Interactive displays educating the general public about glycoscience
Year(s) Of Engagement Activity 2016,2018
URL https://twitter.com/wbturnbull/status/986336659708014593
 
Description ERACoBioTech BioTechHack 2017 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Professional Practitioners
Results and Impact Presentation on the results of our ERASynBio project to an audience of policymakers and other scientists. Also participation in a panel discussion on the future of biotechnology in Europe.
Year(s) Of Engagement Activity 2017
URL https://www.cobiotech.eu/events?event=BioTech_Research_amp_Innovation_Hack_2017
 
Description Pint of Science - Complex Life of Sugars 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach Local
Primary Audience Public/other audiences
Results and Impact Presentation and scientific demonstrations for the general public held in a Leeds pub during Leeds Pint of Science Week
Year(s) Of Engagement Activity 2017
 
Description Reengineering Bacterial Toxins for New Purposes 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Professional Practitioners
Results and Impact dissemination of results at international conferences
Year(s) Of Engagement Activity 2015,2016,2017
 
Description Understanding, Inhibiting and Exploiting Bacterial Toxins 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Type Of Presentation keynote/invited speaker
Geographic Reach International
Primary Audience Professional Practitioners
Results and Impact A research talk outlining the methods we have developed to make site specific chemical modifications on bacterial toxins.

further invitations to talk and to participate in grant applications
Year(s) Of Engagement Activity 2011,2012,2013,2014,2015