Targeting GLP-1 receptor trafficking to improve therapies for type 2 diabetes
Lead Research Organisation:
Imperial College London
Department Name: Metabolism, Digestion and Reproduction
Abstract
Type 2 diabetes (T2D) kills around 4 million people worldwide every year and causes ill health in many more. A major underlying problem in T2D is that specialised cells in the pancreas known as beta cells start to fail and do not produce or secrete enough insulin, which is needed to control blood sugar. Beta cells can be stimulated to release more insulin by activating a surface protein known as glucagon-like peptide-1 receptor (GLP-1R). GLP-1R treatments, known as GLP-1R agonists, are already used in T2D. We have found that a process known as "receptor trafficking" limits the effectiveness of GLP-1R agonists. When the agonist activates the receptor, it also "internalises" it into the cell where it is hidden and can no longer be activated by agonists outside the cell. We showed this using a modified version of a GLP-1R agonist currently used in T2D (exendin-4, also known as "Exenatide"), which we engineered to reduce the internalisation process. This new agonist called "Ex-phe1" was much more effective than unmodified exendin-4 in mice with T2D, as it allowed the receptor to remain longer at the cell surface and be continually reactivated, leading to more insulin release. Also, despite being more effective in beta cells, Ex-phe1 did not cause more nausea, a well-known side effect of GLP-1R agonists.
This project has two goals. The first is to understand how changes in GLP-1R trafficking modify insulin release from beta cells. The second, to be performed in parallel, is to optimise the compounds we generated in our preliminary work to produce new, better, treatments for T2D. Our approach includes experiments in beta cells and pancreatic tissue ("islets"), which allow us to understand events in molecular detail, and in mice with T2D, which tell us what happens at the whole body level. We place a lot of emphasis on using human islets (provided for research when surplus to requirements for organ donation), as we believe this is essential to understand and treat human disease.
We highlight below some key experiments:
1) We will use a combination of computer modelling and genetic modification of the GLP-1R to identify specific interactions between the receptor and Ex-phe1 or other biased GLP-1R agonists which underpin their different trafficking and therapeutic effects. This knowledge will be critical for the design of even more effective drugs in the future.
2) We will genetically modify human islets to reduce levels of "beta-arrestin-2", a protein in beta cells that we believe is a key link between GLP-1R activation, trafficking, and beneficial insulin release. We will also breed genetically modified mice lacking beta-arrestin-2 in beta cells. We will use these systems to look at complex patterns of protein and gene activation within beta cells in response to GLP-1R agonist treatment. From this we hope to identify molecules that could in the future be targeted as new T2D treatments.
3) We will study the long-term effects of our new, improved GLP-1R agonist Ex-phe1 on human islets transplanted into diabetic mice. Importantly, we will use a new approach in which the islets are actually transplanted in the eye, where changes to islet shape, behaviour and overall health can be directly and repeatedly viewed in a microscope. The mice will be treated with a long-lasting formulation of Ex-phe1 or exendin-4 for several months, and we will monitor changes to blood sugar, allowing us to compare the treatments for their effects on overall islet function. Due to well-known differences between mouse and human islets, we believe that using this "humanised" mice is critical to understanding long-term treatment effects.
In summary, we have identified an exciting new approach based on GLP-1R trafficking for improving the treatment of T2D. In this project we will establish the molecular details of why this approach works, identify new potential drug targets and demonstrate the long-term effects of this new treatment in unique humanised mice.
This project has two goals. The first is to understand how changes in GLP-1R trafficking modify insulin release from beta cells. The second, to be performed in parallel, is to optimise the compounds we generated in our preliminary work to produce new, better, treatments for T2D. Our approach includes experiments in beta cells and pancreatic tissue ("islets"), which allow us to understand events in molecular detail, and in mice with T2D, which tell us what happens at the whole body level. We place a lot of emphasis on using human islets (provided for research when surplus to requirements for organ donation), as we believe this is essential to understand and treat human disease.
We highlight below some key experiments:
1) We will use a combination of computer modelling and genetic modification of the GLP-1R to identify specific interactions between the receptor and Ex-phe1 or other biased GLP-1R agonists which underpin their different trafficking and therapeutic effects. This knowledge will be critical for the design of even more effective drugs in the future.
2) We will genetically modify human islets to reduce levels of "beta-arrestin-2", a protein in beta cells that we believe is a key link between GLP-1R activation, trafficking, and beneficial insulin release. We will also breed genetically modified mice lacking beta-arrestin-2 in beta cells. We will use these systems to look at complex patterns of protein and gene activation within beta cells in response to GLP-1R agonist treatment. From this we hope to identify molecules that could in the future be targeted as new T2D treatments.
3) We will study the long-term effects of our new, improved GLP-1R agonist Ex-phe1 on human islets transplanted into diabetic mice. Importantly, we will use a new approach in which the islets are actually transplanted in the eye, where changes to islet shape, behaviour and overall health can be directly and repeatedly viewed in a microscope. The mice will be treated with a long-lasting formulation of Ex-phe1 or exendin-4 for several months, and we will monitor changes to blood sugar, allowing us to compare the treatments for their effects on overall islet function. Due to well-known differences between mouse and human islets, we believe that using this "humanised" mice is critical to understanding long-term treatment effects.
In summary, we have identified an exciting new approach based on GLP-1R trafficking for improving the treatment of T2D. In this project we will establish the molecular details of why this approach works, identify new potential drug targets and demonstrate the long-term effects of this new treatment in unique humanised mice.
Technical Summary
The glucagon-like peptide-1 receptor (GLP-1R) is an important target in type 2 diabetes (T2D) as GLP-1R agonists stimulate beta cell insulin release and promote weight loss. The GLP-1R undergoes rapid agonist-mediated endocytosis, leading us to question whether desensitisation limits GLP-1R agonist treatment effectiveness. We have tested this hypothesis by generating modified versions of the GLP-1R agonist exendin-4. One of these, Ex-phe1, displays a sharply reduced propensity for receptor internalisation, is a more potent insulin secretagogue, and provides enhanced glycaemic control in diabetic mice without increasing nausea, a common side effect of GLP-1R agonists. Ex-phe1 exhibits a distinct pharmacological profile to the parent compound, including fast receptor dissociation kinetics and biased signalling, with beta-arrestin-2 (barr2) recruitment selectively reduced. Here, we will elucidate the molecular mechanisms linking biased GLP-1R activation and trafficking to insulin secretion and examine the therapeutic benefits of biased GLP-1R agonism in a humanised T2D model. Specifically, we will: 1) identify the structural basis for biased GLP-1R activation and trafficking using molecular modelling, reciprocal receptor mutagenesis and a GLP-1R conformational biosensor; 2) determine biased agonist-specific kinomic and transcriptomic responses and changes to interactions with endocytic trafficking regulators in beta cells; 3) confirm the role of barr2 in beta cell responses to GLP-1R agonism both in beta cell-specific barr2 knockout mice and by silencing barr2 in human islets; and 4) establish the long-term effects of biased GLP-1R agonism in a humanised mouse model with human islets implanted into diabetic mice eyes, an approach that uniquely allows serial imaging of islet morphology and function. These studies will provide critical molecular insights into the control of GLP-1R trafficking by biased agonists and will potentially identify new targets for T2D treatment.
Planned Impact
We identify the following groups as likely beneficiaries from work conducted during this project:
1) Patients with T2D and associated metabolic conditions:
In this project, we will develop new molecules that are expected to display improved therapeutic efficacy and tolerability compared to existing treatments. As well as patients with T2D, patients with non-alcoholic fatty liver disease are another key group who are expected to benefit from treatment with biased GLP-1R agonists. GLP-1R agonism is also an investigational strategy for obesity, cardiovascular disease, and neurological diseases. Full clinical development of new agents is expected to take 5-8 years.
2) The NHS:
The S.R.B. group has considerable experience of "first in man" and phase 1 clinical trials. Molecules developed during this project will in the future be taken forward to human studies. The NHS benefits from these studies, for example through Imperial Academic Health Sciences Centre funding. Pre-clinical studies are likely to begin following successful completion of this project.
3) Imperial College London:
We have filed an International (PCT) Patent Application (PCT/GB2017/051045) for biased GLP-1R agonists developed during pilot work for this project. Via Imperial Innovations, a subsidiary of Imperial College London, we will engage with the pharmaceutical industry regarding acquisition of intellectual property. We have a proven track record in this area, for example with the spin-off company Thiakis, which developed the peptide hormone oxyntomodulin as an anti-obesity agent and was subsequently acquired by Wyeth for $150 million.
4) The UK pharmaceutical industry and UK economy:
An estimated 422 million people have diabetes worldwide. Obesity-related comorbidities, such as non-alcoholic fatty liver disease, are at least as prevalent. There is therefore a potentially enormous market for GLP-1R agonists with improved therapeutic efficacy and tolerability, with clear economic benefits for the UK pharmaceutical industry and associated organisations. G.A.R. enjoys extensive interactions with the UK and European pharmaceutical industry as a Work Package leader in the former EU-EFPIA co-funded Innovative Medicines Initiative in Diabetes (IMIDIA) consortium, and presently in the "Rhapsody" consortium under IMI2. The latter involves close collaborations with Servier, Novo Nordsik, Janssen/J&J and Sanofi Aventis, particularly as part of an initiative to identify new circulating biomarkers for T2D development and progression.
5) The wider scientific community:
During this project we will explore a new technological approach to imaging of ligand binding and receptor trafficking in living islets implanted into the anterior chamber of the eye. This would for the first time allow these cellular events to be monitored at cellular resolution in an in vivo environment. Whilst we will focus on the GLP-1R, if this technique is successful, it is likely to be of significant interest to researchers working within the wider GPCR field. Results will be disseminated at scientific conferences and by publication in peer-reviewed journals.
6) The general public:
Our departments have a strong track record of public engagement. We welcome pre-university students into our laboratory for work experience as part of a drive to widen participation. We are involved in the Imperial College Outreach scheme, which organises science education events for school students and their teachers, introducing them to the science of T2D and obesity. We regularly participate in the Imperial Festival, a public engagement event that allows the public to explore and understand our latest research. We also recently contributed to the BBC Horizon programme, "Why Are We Getting So Fat?".
1) Patients with T2D and associated metabolic conditions:
In this project, we will develop new molecules that are expected to display improved therapeutic efficacy and tolerability compared to existing treatments. As well as patients with T2D, patients with non-alcoholic fatty liver disease are another key group who are expected to benefit from treatment with biased GLP-1R agonists. GLP-1R agonism is also an investigational strategy for obesity, cardiovascular disease, and neurological diseases. Full clinical development of new agents is expected to take 5-8 years.
2) The NHS:
The S.R.B. group has considerable experience of "first in man" and phase 1 clinical trials. Molecules developed during this project will in the future be taken forward to human studies. The NHS benefits from these studies, for example through Imperial Academic Health Sciences Centre funding. Pre-clinical studies are likely to begin following successful completion of this project.
3) Imperial College London:
We have filed an International (PCT) Patent Application (PCT/GB2017/051045) for biased GLP-1R agonists developed during pilot work for this project. Via Imperial Innovations, a subsidiary of Imperial College London, we will engage with the pharmaceutical industry regarding acquisition of intellectual property. We have a proven track record in this area, for example with the spin-off company Thiakis, which developed the peptide hormone oxyntomodulin as an anti-obesity agent and was subsequently acquired by Wyeth for $150 million.
4) The UK pharmaceutical industry and UK economy:
An estimated 422 million people have diabetes worldwide. Obesity-related comorbidities, such as non-alcoholic fatty liver disease, are at least as prevalent. There is therefore a potentially enormous market for GLP-1R agonists with improved therapeutic efficacy and tolerability, with clear economic benefits for the UK pharmaceutical industry and associated organisations. G.A.R. enjoys extensive interactions with the UK and European pharmaceutical industry as a Work Package leader in the former EU-EFPIA co-funded Innovative Medicines Initiative in Diabetes (IMIDIA) consortium, and presently in the "Rhapsody" consortium under IMI2. The latter involves close collaborations with Servier, Novo Nordsik, Janssen/J&J and Sanofi Aventis, particularly as part of an initiative to identify new circulating biomarkers for T2D development and progression.
5) The wider scientific community:
During this project we will explore a new technological approach to imaging of ligand binding and receptor trafficking in living islets implanted into the anterior chamber of the eye. This would for the first time allow these cellular events to be monitored at cellular resolution in an in vivo environment. Whilst we will focus on the GLP-1R, if this technique is successful, it is likely to be of significant interest to researchers working within the wider GPCR field. Results will be disseminated at scientific conferences and by publication in peer-reviewed journals.
6) The general public:
Our departments have a strong track record of public engagement. We welcome pre-university students into our laboratory for work experience as part of a drive to widen participation. We are involved in the Imperial College Outreach scheme, which organises science education events for school students and their teachers, introducing them to the science of T2D and obesity. We regularly participate in the Imperial Festival, a public engagement event that allows the public to explore and understand our latest research. We also recently contributed to the BBC Horizon programme, "Why Are We Getting So Fat?".
Organisations
- Imperial College London (Lead Research Organisation, Project Partner)
- Council of Scientific and Industrial Research (CSIR) (Collaboration)
- University College London (Collaboration)
- COVENTRY UNIVERSITY (Collaboration)
- The CNR Institute of Neuroscience, Padova, Italy (Collaboration)
- ImmuPharma (Collaboration)
- IMPERIAL COLLEGE LONDON (Collaboration)
- AstraZeneca (United Kingdom) (Collaboration)
- UNIVERSITY OF OXFORD (Collaboration)
- University of Geneva (Collaboration, Project Partner)
- Rutherford Appleton Laboratory (Collaboration)
- University of Bordeaux (Collaboration)
- Uppsala University (Collaboration)
- Eli Lilly & Company Ltd (Collaboration)
- UNIVERSITY OF BIRMINGHAM (Collaboration)
- Monash University (Collaboration)
- KING'S COLLEGE LONDON (Collaboration)
- University of Alberta (Project Partner)
- University of Pisa (Project Partner)
- Karolinska Institute (Project Partner)
- Imperial College Healthcare NHS Trust (Project Partner)
- Duke University (Project Partner)
- University of Essex (Project Partner)
Publications
Akalestou E
(2020)
Glucocorticoid Metabolism in Obesity and Following Weight Loss.
in Frontiers in endocrinology
Anzilotti C
(2019)
An essential role for the Zn2+ transporter ZIP7 in B cell development.
in Nature immunology
Arcones AC
(2021)
GRK2 regulates GLP-1R-mediated early phase insulin secretion in vivo.
in BMC biology
Ast J
(2022)
Expanded LUXendin Color Palette for GLP1R Detection and Visualization In Vitro and In Vivo.
in JACS Au
Ast J
(2020)
Super-resolution microscopy compatible fluorescent probes reveal endogenous glucagon-like peptide-1 receptor distribution and dynamics.
in Nature communications
Balkhiyarova Z
(2022)
Relationship between glucose homeostasis and obesity in early life-a study of Italian children and adolescents
in Human Molecular Genetics
Buenaventura T
(2019)
Agonist-induced membrane nanodomain clustering drives GLP-1 receptor responses in pancreatic beta cells.
in PLoS biology
Buenaventura T
(2018)
A Targeted RNAi Screen Identifies Endocytic Trafficking Factors That Control GLP-1 Receptor Signaling in Pancreatic ß-Cells.
in Diabetes
Carrat GR
(2020)
The type 2 diabetes gene product STARD10 is a phosphoinositide-binding protein that controls insulin secretory granule biogenesis.
in Molecular metabolism
Cheung R
(2022)
Glucose-Dependent miR-125b Is a Negative Regulator of ß-Cell Function.
in Diabetes
Davies A
(2023)
Appreciating the potential for GPCR crosstalk with ion channels.
in Progress in molecular biology and translational science
Davis SPX
(2019)
Convolutional neural networks for reconstruction of undersampled optical projection tomography data applied to in vivo imaging of zebrafish.
in Journal of biophotonics
De Jesus DS
(2021)
Dysregulation of the Pdx1/Ovol2/Zeb2 axis in dedifferentiated ß-cells triggers the induction of genes associated with epithelial-mesenchymal transition in diabetes.
in Molecular metabolism
Dwivedi OP
(2019)
Loss of ZnT8 function protects against diabetes by enhanced insulin secretion.
in Nature genetics
El Eid L
(2022)
Biased agonism and polymorphic variation at the GLP-1 receptor: Implications for the development of personalised therapeutics.
in Pharmacological research
Fang Z
(2020)
The Influence of Peptide Context on Signaling and Trafficking of Glucagon-like Peptide-1 Receptor Biased Agonists.
in ACS pharmacology & translational science
Fang Z
(2020)
Ligand-Specific Factors Influencing GLP-1 Receptor Post-Endocytic Trafficking and Degradation in Pancreatic Beta Cells.
in International journal of molecular sciences
Fine NHF
(2018)
Glucocorticoids Reprogram ß-Cell Signaling to Preserve Insulin Secretion.
in Diabetes
Fremaux J
(2019)
Ureidopeptide GLP-1 analogues with prolonged activity in vivo via signal bias and altered receptor trafficking.
in Chemical science
Georgiadou E
(2020)
Age matters: Grading granule secretion in beta cells.
in The Journal of biological chemistry
Georgiadou E
(2020)
Control by Ca2+ of mitochondrial structure and function in pancreatic ß-cells.
in Cell calcium
Gromada J
(2018)
The a-cell in diabetes mellitus.
in Nature reviews. Endocrinology
Hamilton A
(2018)
Adrenaline Stimulates Glucagon Secretion by Tpc2-Dependent Ca2+ Mobilization From Acidic Stores in Pancreatic a-Cells.
in Diabetes
Hinds CE
(2023)
Abolishing ß-arrestin recruitment is necessary for the full metabolic benefits of G protein-biased glucagon-like peptide-1 receptor agonists.
in Diabetes, obesity & metabolism
Hinds CE
(2021)
A glucagon analogue decreases body weight in mice via signalling in the liver.
in Scientific reports
Hu M
(2020)
Functional Genomics in Pancreatic ß Cells: Recent Advances in Gene Deletion and Genome Editing Technologies for Diabetes Research.
in Frontiers in endocrinology
Jaafar R
(2019)
mTORC1 to AMPK switching underlies ß-cell metabolic plasticity during maturation and diabetes.
in The Journal of clinical investigation
Jones B
(2018)
Targeting GLP-1 receptor trafficking to improve agonist efficacy.
in Nature communications
Jones B
(2022)
The therapeutic potential of GLP-1 receptor biased agonism.
in British journal of pharmacology
Jones B
(2022)
The Metabolomic Effects of Tripeptide Gut Hormone Infusion Compared to Roux-en-Y Gastric Bypass and Caloric Restriction.
in The Journal of clinical endocrinology and metabolism
Jones B
(2022)
In vivo and in vitro characterization of GL0034, a novel long-acting glucagon-like peptide-1 receptor agonist.
in Diabetes, obesity & metabolism
Jones B
(2021)
Genetic and biased agonist-mediated reductions in ß-arrestin recruitment prolong cAMP signaling at glucagon family receptors.
in The Journal of biological chemistry
Khan R
(2020)
Effects on pancreatic Beta and other Islet cells of the glucose-dependent insulinotropic polypeptide.
in Peptides
Kleiner S
(2018)
Mice harboring the human SLC30A8 R138X loss-of-function mutation have increased insulin secretory capacity.
in Proceedings of the National Academy of Sciences of the United States of America
Lucey M
(2021)
Acylation of the Incretin Peptide Exendin-4 Directly Impacts Glucagon-Like Peptide-1 Receptor Signaling and Trafficking.
in Molecular pharmacology
Lucey M
(2020)
Disconnect between signalling potency and in vivo efficacy of pharmacokinetically optimised biased glucagon-like peptide-1 receptor agonists.
in Molecular metabolism
| Title | "Enhanced endosomal signaling and desensitization of GLP-1R versus GIPR in pancreatic beta cells" - Supplementary data |
| Description | Supplementary data |
| Type Of Art | Film/Video/Animation |
| Year Produced | 2023 |
| URL | https://figshare.com/articles/figure/_Enhanced_endosomal_signaling_and_desensitization_of_GLP-1R_ver... |
| Title | "Enhanced endosomal signaling and desensitization of GLP-1R versus GIPR in pancreatic beta cells" - Supplementary data |
| Description | Supplementary data |
| Type Of Art | Film/Video/Animation |
| Year Produced | 2023 |
| URL | https://figshare.com/articles/figure/_Enhanced_endosomal_signaling_and_desensitization_of_GLP-1R_ver... |
| Description | Academy of Medical Sciences "Studying the impact of biased GLP-1 receptor signalling in humans" |
| Amount | £30,000 (GBP) |
| Funding ID | SGL019\1078 |
| Organisation | Academy of Medical Sciences (AMS) |
| Sector | Charity/Non Profit |
| Country | United Kingdom |
| Start | 08/2018 |
| End | 08/2019 |
| Description | DUK PhD Studentship: "Spatiotemporal Control of Signalling of the GLP-1R Variant Ala316Thr in Pancreatic Beta Cells" |
| Amount | £199,079 (GBP) |
| Funding ID | 19/0006094 |
| Organisation | Diabetes UK |
| Sector | Charity/Non Profit |
| Country | United Kingdom |
| Start | 01/2021 |
| End | 12/2023 |
| Description | Functional Studies on Novel GLP-1R Agonists |
| Amount | £232,800 (GBP) |
| Organisation | Sun Pharmaceuticals |
| Sector | Private |
| Country | India |
| Start | 02/2019 |
| End | 01/2021 |
| Description | Integrated Biological Imaging Network (IBIN) Pump-priming Project Application |
| Amount | £13,642 (GBP) |
| Organisation | United Kingdom Research and Innovation |
| Sector | Public |
| Country | United Kingdom |
| Start | 06/2022 |
| End | 09/2022 |
| Description | LRAP - The effects of ligand, genotype and tissue diversity on GIPR trafficking responses |
| Amount | $264,995 (USD) |
| Organisation | Eli Lilly & Company Ltd |
| Sector | Private |
| Country | United Kingdom |
| Start | 10/2021 |
| End | 10/2023 |
| Description | Real-time imaging of cyclic AMP dynamics in pancreatic islets to understand the role of beta-arrestin-2 in insulin secretion |
| Amount | £9,681 (GBP) |
| Organisation | Society for Endocrinology |
| Sector | Charity/Non Profit |
| Country | United Kingdom |
| Start | 01/2021 |
| End | 12/2021 |
| Description | Regulation of beta cell incretin responses by the lipid microenvironment |
| Amount | £318,350 (GBP) |
| Funding ID | NGCS-2020-451 |
| Organisation | Government of the UK |
| Department | Commonwealth Scholarship Commission |
| Sector | Public |
| Country | United Kingdom |
| Start | 01/2021 |
| End | 12/2023 |
| Description | Society for Endocrinology: "Proof of concept for personalisation of GLP-1 receptor agonist treatment using biased ligands" |
| Amount | £10,000 (GBP) |
| Organisation | Society for Endocrinology |
| Sector | Charity/Non Profit |
| Country | United Kingdom |
| Start | 08/2018 |
| End | 08/2019 |
| Description | Understanding the tissue-specific effects of biased GLP-1 receptor agonism in type 2 diabetes |
| Amount | € 99,000 (EUR) |
| Organisation | European Association for the Study of Diabetes (EASD) |
| Sector | Charity/Non Profit |
| Country | Germany |
| Start | 01/2020 |
| End | 12/2021 |
| Title | Biased GLP-1R agonists with improved therapeutic potential |
| Description | Lipid conjugated exendin-4, exendin-phe1, exendin-asp3 |
| Type Of Material | Biological samples |
| Year Produced | 2018 |
| Provided To Others? | No |
| Impact | None yet |
| Title | Fluorescent conjugates of biased GLP-1R agonists for cellular and tissue imaging |
| Description | Fluorescent agonists for cellular and tissue imaging |
| Type Of Material | Biological samples |
| Year Produced | 2019 |
| Provided To Others? | No |
| Impact | Analysis of brain entry and single molecule localisation microscopy |
| Title | Further GLP-1R-active peptides for evaluation of modes of receptor binding |
| Description | GLP-1/exendin-4 chimeric compounds |
| Type Of Material | Biological samples |
| Year Produced | 2018 |
| Provided To Others? | No |
| Impact | None yet |
| Title | GLP-1R A316T mouse |
| Description | humanised GLP-1R mouse with A316T SNP |
| Type Of Material | Model of mechanisms or symptoms - mammalian in vivo |
| Year Produced | 2022 |
| Provided To Others? | No |
| Impact | assessment of A316T mutation effects in vivo |
| Title | HALO-GIPR mouse |
| Description | mouse with GIPR endogenously tagged with HALO |
| Type Of Material | Model of mechanisms or symptoms - mammalian in vivo |
| Year Produced | 2022 |
| Provided To Others? | No |
| Impact | assessment of GIPR expression profile in vivo |
| Title | INS SNAP-GLP-1R C438A and INS SNAP-GLP-1R A316T |
| Description | Beta cell lines expressing mutant SNAP-GLP-1Rs on a GLP-1R -/- background |
| Type Of Material | Cell line |
| Year Produced | 2018 |
| Provided To Others? | No |
| Impact | Investigation of the effects of point mutations in GLP-1R trafficking and signalling |
| Title | Lentiviral CRISPR constructs for zDHHC family of S-acyltransferases |
| Description | Lentiviral CRISPR constructs for generation of KO cell lines for zDHHC5, 20 and 21 |
| Type Of Material | Biological samples |
| Year Produced | 2019 |
| Provided To Others? | No |
| Impact | Generation of zDHHC5, 20 and 21 -/- mouse beta cell lines |
| Title | SNAP-GLP-1R A316T |
| Description | SNAP-GLP-1R with A316T single nucleotide polymorphism |
| Type Of Material | Biological samples |
| Year Produced | 2018 |
| Provided To Others? | No |
| Impact | Investigation of the effect of SNPs on GLP-1R trafficking and signalling |
| Title | SNAP-receptor-PK plasmids |
| Description | SNAP-GLP-1R/GIPR/GCGR tagged with Prolink (PK) |
| Type Of Material | Biological samples |
| Year Produced | 2018 |
| Provided To Others? | No |
| Impact | Analysis of beta arrestin recruitment by enzyme fragment complementation assay |
| Title | SNAP-tagged receptor beta cell lines |
| Description | SNAP-GLP-1R / SNAP-GIPR-expressing beta cell lines in GLP-1R/GIPR -/- background |
| Type Of Material | Cell line |
| Year Produced | 2019 |
| Provided To Others? | No |
| Impact | Use of these cell lines to investigate the effect of receptor SNPs and dual GLP-1R/GIPR agonists |
| Title | Targeted cAMP biosensors |
| Description | T-Epac-VV cAMP biosensor tagged with lipid raft, endosomal and TGN localisation signals |
| Type Of Material | Biological samples |
| Year Produced | 2018 |
| Provided To Others? | No |
| Impact | Measurement of cAMP from subcellular compartments |
| Title | Gene expression dataset |
| Description | Transcriptomic data of gene expression levels following GLP-1R biased agonist stimulation of beta cells and hypothalamic neuronal cells |
| Type Of Material | Database/Collection of data |
| Year Produced | 2019 |
| Provided To Others? | No |
| Impact | None yet |
| Title | Kinase activation dataset |
| Description | Kinomic data of kinase activity levels following GLP-1R biased agonist stimulation of beta cells |
| Type Of Material | Database/Collection of data |
| Year Produced | 2019 |
| Provided To Others? | No |
| Impact | None yet |
| Description | Analysis of GLP-1R agonist binding cooperativity and receptor oligomerisation |
| Organisation | Imperial College London |
| Department | Institute of Reproductive and Developmental Biology |
| Country | United Kingdom |
| Sector | Academic/University |
| PI Contribution | We are developing a number of assays to analyse differences in binding cooperativity between GLP-1 receptor agonists |
| Collaborator Contribution | Expertise on BRET assays and oligomerisation analysis |
| Impact | Multidisciplinary: pharmacology/cell biology/endocrinology |
| Start Year | 2017 |
| Description | Analysis of GLP-1R agonist binding cooperativity and receptor oligomerisation |
| Organisation | Imperial College London |
| Department | School of Public Health |
| Country | United Kingdom |
| Sector | Academic/University |
| PI Contribution | We are developing a number of assays to analyse differences in binding cooperativity between GLP-1 receptor agonists |
| Collaborator Contribution | Expertise on BRET assays and oligomerisation analysis |
| Impact | Multidisciplinary: pharmacology/cell biology/endocrinology |
| Start Year | 2017 |
| Description | Analysis of GLP-1R trafficking differences in response to a panel of exendin-4 analogues with single amino acid substitutions |
| Organisation | Imperial College London |
| Department | Section of Endocrinology and Investigative Medicine |
| Country | United Kingdom |
| Sector | Academic/University |
| PI Contribution | Analysis of GLP-1R trafficking in response to a panel of biased exendin-4 analogues |
| Collaborator Contribution | Panel of exendin-4 analogues exhibiting varying degrees of GLP-1R biased agonism |
| Impact | Multi-disciplinary: Pharmacology/Cell Biology On-going partnership that might provide a novel therapeutic approach to T2DM Paper submitted and under review Biased agonists now patented (ICL) |
| Start Year | 2015 |
| Description | Analysis of biased agonist GLP-1R responses in the choroid plexus |
| Organisation | University of Birmingham |
| Department | Institute of Metabolism and Systems Research (IMSR) |
| Country | United Kingdom |
| Sector | Academic/University |
| PI Contribution | Plasmids, lentiviruses, modified agonists |
| Collaborator Contribution | Choroid samples and ICV/choroid plexus techniques |
| Impact | No outputs yet |
| Start Year | 2019 |
| Description | Analysis of biased agonist brain responses |
| Organisation | University College London |
| Department | Division of Biosciences |
| Country | United Kingdom |
| Sector | Academic/University |
| PI Contribution | Brain samples from biased agonist treated mice |
| Collaborator Contribution | Analysis of neuronal activity |
| Impact | No outputs yet |
| Start Year | 2019 |
| Description | Analysis of chemically-modified GLP-1 analogues |
| Organisation | ImmuPharma |
| Department | UREKA |
| Country | United Kingdom |
| Sector | Private |
| PI Contribution | Analysis of GLP-1R trafficking and signalling bias from chemically-modified GLP-1 analogues |
| Collaborator Contribution | Chemically-modified GLP-1 analogues |
| Impact | No outputs yet |
| Start Year | 2018 |
| Description | Analysis of chemically-modified GLP-1 analogues |
| Organisation | University of Bordeaux |
| Country | France |
| Sector | Academic/University |
| PI Contribution | Analysis of GLP-1R trafficking and signalling bias from chemically-modified GLP-1 analogues |
| Collaborator Contribution | Chemically-modified GLP-1 analogues |
| Impact | No outputs yet |
| Start Year | 2018 |
| Description | Analysis of incretin receptor expression and function in zebrafish |
| Organisation | University of Geneva |
| Department | Department of Biochemistry |
| Country | Switzerland |
| Sector | Academic/University |
| PI Contribution | Fluorescent GLP-1R agonists and SNAP-tagged expression vectors |
| Collaborator Contribution | expertise in the generation of transgenic zebrafish lines, labelling and localisation of dual GLP-1R/GCGR in zebrafish embryos |
| Impact | No outputs to date |
| Start Year | 2019 |
| Description | Analysis of receptor clustering by FCS microscopy |
| Organisation | Imperial College London |
| Department | National Heart & Lung Institute (NHLI) |
| Country | United Kingdom |
| Sector | Academic/University |
| PI Contribution | study of class B GPCR clustering |
| Collaborator Contribution | FCS data collection and analysis |
| Impact | co-authored publications |
| Start Year | 2019 |
| Description | Consultancy agreement |
| Organisation | ImmuPharma |
| Department | UREKA |
| Country | United Kingdom |
| Sector | Private |
| PI Contribution | Consultancy services |
| Collaborator Contribution | Salary contribution |
| Impact | N/A |
| Start Year | 2021 |
| Description | Cryo-CLEM and MINFLUX super-resolution analysis of GLP-1R behaviours |
| Organisation | Rutherford Appleton Laboratory |
| Department | Central Laser Facility |
| Country | United Kingdom |
| Sector | Academic/University |
| PI Contribution | High resolution microscopy to localise GLP-1R to membrane contact sites and investigate receptor oligomerisation at the plasma membrane |
| Collaborator Contribution | consimables, technical expertise and microscope access |
| Impact | preliminary data |
| Start Year | 2022 |
| Description | Development of novel tools to study GPCR palmitoylation |
| Organisation | Imperial College London |
| Country | United Kingdom |
| Sector | Academic/University |
| PI Contribution | Genetically modified beta cell lines (CRISPR/Cas9) |
| Collaborator Contribution | Plasmids and tools to engineer cell lines to screen for specific palmitoyltransferase partners in beta cells |
| Impact | N/A |
| Start Year | 2019 |
| Description | Generation of fluorescently labelled GLP-1R agonists |
| Organisation | Imperial College London |
| Department | Department of Chemistry |
| Country | United Kingdom |
| Sector | Academic/University |
| PI Contribution | Testing fluorescently labelled GLP-1R agonists for a variety of applications Modified agonists and fluorophores |
| Collaborator Contribution | Chemical sysnthesis of fluorescently labelled GLP-1R agonists |
| Impact | No outputs yet |
| Start Year | 2018 |
| Description | Investigation of in vivo effects of biased agonists in eye-implanted pancreatic islets |
| Organisation | Imperial College London |
| Department | Department of Medicine |
| Country | United Kingdom |
| Sector | Academic/University |
| PI Contribution | Biased agonists and expertise in in vivo evaluation of incretin signalling |
| Collaborator Contribution | mouse model of islets implantation and imaging from anterior chamber of the eye |
| Impact | None yet |
| Start Year | 2019 |
| Description | Lipidomic analysis of GLP-1R biased agonist signalling |
| Organisation | University of Geneva |
| Department | Department of Biochemistry |
| Country | Switzerland |
| Sector | Academic/University |
| PI Contribution | Pancreatic beta cell and neuronal samples stimulated with biased GLP-1R agonists |
| Collaborator Contribution | Analysis of sample lipid species composition |
| Impact | Data on lipid composition downstream of GLP-1R signalling |
| Start Year | 2019 |
| Description | Mass spectrometry analysis of GLP-1R biased agonist signalling |
| Organisation | Imperial College London |
| Country | United Kingdom |
| Sector | Academic/University |
| PI Contribution | Preparation of mass spec samples with factors pulled down with GLP-1R following stimulation with biased agonists in pancreatic beta cells |
| Collaborator Contribution | Mass spec analysis of GLP-1R binding partners |
| Impact | Datasets of binding partners |
| Start Year | 2019 |
| Description | Mass spectrometry analysis of GLP-1R interactions downstream of biased agonist stimulation |
| Organisation | Imperial College London |
| Department | Department of Chemistry |
| Country | United Kingdom |
| Sector | Academic/University |
| PI Contribution | Pulled-down samples of SNAP-GLP-1R from pancreatic beta cells |
| Collaborator Contribution | Mass spectrometry sample preparation and analysis |
| Impact | No outputs yet |
| Start Year | 2018 |
| Description | Mathematical modelling of human data |
| Organisation | The CNR Institute of Neuroscience, Padova, Italy |
| Country | Italy |
| Sector | Academic/University |
| PI Contribution | Human physiological study data |
| Collaborator Contribution | Study of incretin effects in healthy and type 2 diabetic subjects through the use of mathematical models |
| Impact | No outputs yet |
| Start Year | 2018 |
| Description | Molecular dynamics simulations of class B GPCR interactions with cholesterol |
| Organisation | Imperial College London |
| Country | United Kingdom |
| Sector | Academic/University |
| PI Contribution | Testing GLP-1R mutants fro potential cholesterol binding sites |
| Collaborator Contribution | Prediction of cholesterol binding sites for the GLP-1R |
| Impact | Predicted cholesterol binding sites for the GLP-1R |
| Start Year | 2019 |
| Description | Molecular dynamics simulations of class B GPCR interactions with cholesterol |
| Organisation | University of Oxford |
| Department | Department of Biochemistry |
| Country | United Kingdom |
| Sector | Academic/University |
| PI Contribution | Testing GLP-1R mutants fro potential cholesterol binding sites |
| Collaborator Contribution | Prediction of cholesterol binding sites for the GLP-1R |
| Impact | Predicted cholesterol binding sites for the GLP-1R |
| Start Year | 2019 |
| Description | Molecular dynamics simulations of class B GPCR interactions with the lipid environment |
| Organisation | Council of Scientific and Industrial Research (CSIR) |
| Department | Centre for Cellular and Molecular Biology (CCMB) |
| Country | India |
| Sector | Academic/University |
| PI Contribution | Experimental validation of simulation results |
| Collaborator Contribution | In silico analysis of GLP-1R-lipid interactions |
| Impact | No outputs yet |
| Start Year | 2019 |
| Description | Molecular dynamics simulations of class B GPCRs |
| Organisation | Coventry University |
| Country | United Kingdom |
| Sector | Academic/University |
| PI Contribution | study the effect of biased agonists on receptor function |
| Collaborator Contribution | molecualr dinamics simulations of GLP-1R |
| Impact | co-authored publications |
| Start Year | 2019 |
| Description | Sharing of materials and knowledge for the investigation of biased GLP-1R/GIPR agonists |
| Organisation | AstraZeneca |
| Department | MedImmune |
| Country | United Kingdom |
| Sector | Private |
| PI Contribution | Generation of cell lines and analysis of GLP-1R/GIPR biology |
| Collaborator Contribution | Cell lines Antibodies |
| Impact | Tagged receptor cell sublines |
| Start Year | 2018 |
| Description | Sharing of materials and knowledge for the investigation of biased class B GPCR agonists |
| Organisation | Eli Lilly & Company Ltd |
| Country | United Kingdom |
| Sector | Private |
| PI Contribution | Spatiotenporal regulation of GCGR signalling |
| Collaborator Contribution | GCGR blocking antibodies and control IgGs |
| Impact | no outputs so far |
| Start Year | 2020 |
| Description | Single particle tracking of biased agonist-stimulated GLP-1Rs |
| Organisation | University of Birmingham |
| Department | Institute of Metabolism and Systems Research (IMSR) |
| Country | United Kingdom |
| Sector | Academic/University |
| PI Contribution | Cell lines, plasmids and biased agonists |
| Collaborator Contribution | SPT tracking TIRFM and image analysis |
| Impact | No outputs yet |
| Start Year | 2019 |
| Description | Single-molecule localisation microscopy analysis of plasma membrane GPCR dynamics |
| Organisation | University of Birmingham |
| Country | United Kingdom |
| Sector | Academic/University |
| PI Contribution | Preparation of samples and labelling |
| Collaborator Contribution | Imaging and data analysis |
| Impact | Joint publication: Agonist-induced membrane nanodomain clustering drives GLP-1 receptor responses in pancreatic beta cells. PLoS Biol. 2019 Aug 20;17(8):e3000097. doi: 10.1371/journal.pbio.3000097. |
| Start Year | 2019 |
| Description | Structural analysis of GLP-1R-biased agonist complex |
| Organisation | Monash University |
| Country | Australia |
| Sector | Academic/University |
| PI Contribution | Biased agonists |
| Collaborator Contribution | Expertise in structural biology |
| Impact | No outputs yet |
| Start Year | 2019 |
| Description | Study of GIPR mutants and development of new GIPR in vivo tools |
| Organisation | Eli Lilly & Company Ltd |
| Department | Neuroscience Eli Lilly |
| Country | United States |
| Sector | Private |
| PI Contribution | Experiments and data analysis |
| Collaborator Contribution | Funding and mouse lines |
| Impact | N/A |
| Start Year | 2021 |
| Description | Study of GLP-1R control of membrane contact sites |
| Organisation | Uppsala University |
| Country | Sweden |
| Sector | Academic/University |
| PI Contribution | examination of control of MCS formation by the GLP-1R |
| Collaborator Contribution | Tools to examine MCS formation in beta cells |
| Impact | Preliminary data |
| Start Year | 2022 |
| Description | Superresolution STED-FCS technology pump-priming |
| Organisation | King's College London |
| Country | United Kingdom |
| Sector | Academic/University |
| PI Contribution | STED-FCS technology |
| Collaborator Contribution | Funding |
| Impact | N/A |
| Start Year | 2022 |
| Description | Transcriptome and kinome analysis of GLP-1R biased agonist signalling |
| Organisation | Imperial College London |
| Department | Department of Medicine |
| Country | United Kingdom |
| Sector | Academic/University |
| PI Contribution | Data on GLP-1R trafficking and signalling differences |
| Collaborator Contribution | RNA seq and kinome analysis |
| Impact | No output yet |
| Start Year | 2017 |
| Description | Transcriptome and kinome analysis of GLP-1R biased agonist signalling |
| Organisation | Imperial College London |
| Department | Department of Paediatrics |
| Country | United Kingdom |
| Sector | Academic/University |
| PI Contribution | Data on GLP-1R trafficking and signalling differences |
| Collaborator Contribution | RNA seq and kinome analysis |
| Impact | No output yet |
| Start Year | 2017 |
| Title | "RETRO study" |
| Description | "RETRO study" - a human physiological study investigating the effects of biased GLP-1R agonists in humans. Academy of Medical Sciences. "Studying the impact of biased GLP-1 receptor signalling in humans". Ben Jones (PI), £30,000. 08/2018 - 08/2019 Please note this is a physiological study rather than a Clinical Trial of an Investigational Medicinal Product |
| Type | Therapeutic Intervention - Drug |
| Current Stage Of Development | Refinement. Clinical |
| Year Development Stage Completed | 2018 |
| Development Status | Under active development/distribution |
| Impact | Human physiological study investigating the effects of biased GLP-1R agonists in humans. |
| Description | COMPARE Workshop on Single-Molecule imaging and spectroscopy of membrane proteins |
| Form Of Engagement Activity | Participation in an activity, workshop or similar |
| Part Of Official Scheme? | No |
| Geographic Reach | National |
| Primary Audience | Professional Practitioners |
| Results and Impact | Participation in COMPARE Workshop on Single-Molecule imaging and spectroscopy of membrane proteins |
| Year(s) Of Engagement Activity | 2019 |
| Description | Creation of the Imperial College Cross-Faculty Membrane Receptor Network |
| Form Of Engagement Activity | A formal working group, expert panel or dialogue |
| Part Of Official Scheme? | No |
| Geographic Reach | National |
| Primary Audience | Professional Practitioners |
| Results and Impact | Creation of a Network of researchers working on different aspects of membrane receptors |
| Year(s) Of Engagement Activity | 2022,2023 |
| URL | https://www.imperial.ac.uk/imperial-membrane-receptor-network-of-excellence/ |
| Description | Session chair, Diabetes UK Professional Conference |
| Form Of Engagement Activity | Participation in an activity, workshop or similar |
| Part Of Official Scheme? | No |
| Geographic Reach | National |
| Primary Audience | Professional Practitioners |
| Results and Impact | Session chair, Diabetes UK Professional Conference |
| Year(s) Of Engagement Activity | 2019 |