Investigating the role of immature beta cells in insulin release from the intact islet
Lead Research Organisation:
University of Birmingham
Department Name: Inst of Metabolism & Systems Research
Abstract
Type 2 diabetes mellitus (T2DM) occurs when pancreatic beta-cells are unable to release enough insulin to combat resistance to the hormone. The resulting increase in blood glucose levels drives a range of severe complications including cardiovascular disease, renal and liver failure, retinal degeneration and cancer. As such T2DM is considered an immediate healthcare crisis, affecting 1 in 10 adults in the UK and consuming 9% of the NHS annual budget. Despite intensive research efforts, T2DM incidence continues to rise and NHS spend is projected to double by 2035. To have any hope of halting this trend, we urgently need to open up new research avenues to identify mechanisms and therapeutic targets. Research from us and others over the past few years has shown that not all beta-cells are equal. Much like society, beta-cells possess different ages, abilities and capacities. When the proportions between these different beta-cells become imbalanced, for example during obesity, ageing or diabetes, then insulin release declines. In particular, predominance of younger or immature beta-cells is associated with generation of more beta-cells, but at the expense of insulin release. Despite this, practically nothing is known about the immature beta-cells that are normally resident in the healthy adult pancreas. Our most recent studies have shown that, contrary to expectation, immature beta-cells might in fact play a central role in regulating insulin release. We now aim to:
1) investigate how loss of immature cells influences behaviour of the other, more mature beta-cells;
2) control the maturity status of individual beta-cells using light to directly explore their contribution to insulin release;
3) develop a mouse model where immature beta-cells can be made to be more mature using drugs in the food;
4) determine whether loss of immature beta-cells predisposes to T2DM.
It is anticipated that these studies will provide an updated blueprint for insulin release, with relevance for T2DM treatment as well as the generation of islets from stem cells for transplantation.
1) investigate how loss of immature cells influences behaviour of the other, more mature beta-cells;
2) control the maturity status of individual beta-cells using light to directly explore their contribution to insulin release;
3) develop a mouse model where immature beta-cells can be made to be more mature using drugs in the food;
4) determine whether loss of immature beta-cells predisposes to T2DM.
It is anticipated that these studies will provide an updated blueprint for insulin release, with relevance for T2DM treatment as well as the generation of islets from stem cells for transplantation.
Technical Summary
It is becoming increasingly apparent that pancreatic beta-cells can be grouped into subpopulations according to their transcriptomic, protein and functional signatures. A unifying feature is that immature and mature beta-cell subpopulations co-exist within the islet. Plasticity in these subpopulations occurs during ageing and metabolic stress, and is generally associated with reductions in insulin release. In particular, maturity might be traded for proliferation at the expense of insulin secretion. However, this questions the role of immature beta-cells in the adult islet given the low levels of beta-cell proliferation and the requirement to maintain function. Moreover, most studies to date have either used isolated beta-cells, devoid of paracrine, juxtacrine and electrical communications known to shape beta-cell function, or alternatively have averaged measures across the population. Our preliminary data show that loss of immature beta-cells from the islet leads to insulin secretory failure due to impaired cell-cell communication, metabolism and amplifying signals. As such, we hypothesise that immature beta-cells are an underappreciated player in the regulation of insulin release when viewed directly within the islet setting. By melding cutting-edge imaging, recombinant genetics, light-activated transcription factors and photopharmacology, we now aim to functionally interrogate a role for immature beta-cells in islet operation and insulin release. Specifically, we will:
1) produce and test a loss-of-immaturity beta-cell model using viral approaches and mouse genetics;
2) deploy next-generation optogenes and photoactivatable doxycycline to reversibly control beta-cell maturity with pinpoint precision in multiple systems;
3) understand how inducible loss of immature beta-cells in vivo might influence glucose tolerance;
4) investigate whether loss of immature beta-cells leads to islet failure and diabetes during metabolic stress.
1) produce and test a loss-of-immaturity beta-cell model using viral approaches and mouse genetics;
2) deploy next-generation optogenes and photoactivatable doxycycline to reversibly control beta-cell maturity with pinpoint precision in multiple systems;
3) understand how inducible loss of immature beta-cells in vivo might influence glucose tolerance;
4) investigate whether loss of immature beta-cells leads to islet failure and diabetes during metabolic stress.
Planned Impact
Pharmaceutical industry: The proposed research will provide new mechanistic insights into how beta-cell heterogeneity within the islet contributes to insulin release. This may lead to the identification of drug targets achieving glucose homeostasis in humans. Indeed, current treatment regimens are based on single cell characteristics, yet increasing evidence suggests that we should be aiming to reverse "networkopathies" (i.e. defects in the functional pattern of cell gene expression/activation). Moreover, islets engineered from stem cells (iPSC) are currently in phase 1 trials for transplantation during diabetes, but their in vitro performance does not match that of native islets. The present studies will therefore be important for informing how we can engineer better performing and more robust iPSC-derived islets for transplantation.
UK economy: Since the diabetes drugs market is worth billions per year, and the stem cell market predicted to be similarly large, the licensing or spin-off of any molecules/novel targets may contribute to UK economic competitiveness.
Life Sciences suppliers: We will generate a number of novel tools to precisely induce gene expression in single cells within a tissue using light. This will impact many different research fields (cancer, cardiovascular, neuroscience) that currently rely on less specific methodology to control gene expression levels.
Patients living with diabetes: The characterization of novel mechanisms governing insulin secretion from islets may eventually lead to therapies which allow better or ancillary control of blood glucose levels (e.g. by preserving the balance between immature and mature beta-cells). Moreover, by applying this information to the production of islets from iPSC cells, transplants may be more robust and perform better.
Staff: The postdoctoral researcher and technician will be recruited into the IMSR (~ 80 researchers), providing excellent opportunities for multi-disciplinary training. The present project proposal seeks to apply cutting-edge technologies to functionally dissect the contribution of immature beta-cells to insulin secretion. This would equip the postdoctoral researcher with a broad and interdisciplinary skills-set, which would open up the possibility to work in a number of other academic fields, thus increasing their future career prospects. In addition, such a background is desirable for the private sector. For example, PhD candidates with strong interdisciplinary skills are attractive not only to the pharmaceutical industry, but also to the financial sector.
General engagement: Societal impact will be assisted through engagement activities. These include formal and informal presentations (e.g. Pint of Science, Life Sciences in Six), press releases and social media (e.g. Twitter). The lab has an extensive track record in public engagement, including production of bitesize YouTube videos and appearances on BBC Radio.
UK economy: Since the diabetes drugs market is worth billions per year, and the stem cell market predicted to be similarly large, the licensing or spin-off of any molecules/novel targets may contribute to UK economic competitiveness.
Life Sciences suppliers: We will generate a number of novel tools to precisely induce gene expression in single cells within a tissue using light. This will impact many different research fields (cancer, cardiovascular, neuroscience) that currently rely on less specific methodology to control gene expression levels.
Patients living with diabetes: The characterization of novel mechanisms governing insulin secretion from islets may eventually lead to therapies which allow better or ancillary control of blood glucose levels (e.g. by preserving the balance between immature and mature beta-cells). Moreover, by applying this information to the production of islets from iPSC cells, transplants may be more robust and perform better.
Staff: The postdoctoral researcher and technician will be recruited into the IMSR (~ 80 researchers), providing excellent opportunities for multi-disciplinary training. The present project proposal seeks to apply cutting-edge technologies to functionally dissect the contribution of immature beta-cells to insulin secretion. This would equip the postdoctoral researcher with a broad and interdisciplinary skills-set, which would open up the possibility to work in a number of other academic fields, thus increasing their future career prospects. In addition, such a background is desirable for the private sector. For example, PhD candidates with strong interdisciplinary skills are attractive not only to the pharmaceutical industry, but also to the financial sector.
General engagement: Societal impact will be assisted through engagement activities. These include formal and informal presentations (e.g. Pint of Science, Life Sciences in Six), press releases and social media (e.g. Twitter). The lab has an extensive track record in public engagement, including production of bitesize YouTube videos and appearances on BBC Radio.
Publications
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
Mendive-Tapia L
(2023)
Acid-Resistant BODIPY Amino Acids for Peptide-Based Fluorescence Imaging of GPR54 Receptors in Pancreatic Islets
in Angewandte Chemie
Mendive-Tapia L
(2023)
Acid-Resistant BODIPY Amino Acids for Peptide-Based Fluorescence Imaging of GPR54 Receptors in Pancreatic Islets.
in Angewandte Chemie (International ed. in English)
Marzook A
(2021)
Evaluation of efficacy- versus affinity-driven agonism with biased GLP-1R ligands P5 and exendin-F1.
in Biochemical pharmacology
Pickford P
(2020)
Signalling, trafficking and glucoregulatory properties of glucagon-like peptide-1 receptor agonists exendin-4 and lixisenatide.
in British journal of pharmacology
Hu M
(2021)
Chromatin 3D interaction analysis of the STARD10 locus unveils FCHSD2 as a regulator of insulin secretion.
in Cell reports
Westbrook RL
(2022)
Proline synthesis through PYCR1 is required to support cancer cell proliferation and survival in oxygen-limiting conditions.
in Cell reports
Yip SH
(2019)
Elevated Prolactin during Pregnancy Drives a Phenotypic Switch in Mouse Hypothalamic Dopaminergic Neurons.
in Cell reports
Viloria K
(2020)
Vitamin-D-Binding Protein Contributes to the Maintenance of a Cell Function and Glucagon Secretion.
in Cell reports
Cadilhac C
(2021)
Excitatory granule neuron precursors orchestrate laminar localization and differentiation of cerebellar inhibitory interneuron subtypes.
in Cell reports
Title | Artist in residence |
Description | We are currently working with an artist in residence who will design a creative media piece to show how diversity across beta cells impacts insulin release. |
Type Of Art | Artwork |
Year Produced | 2020 |
Impact | The artwork is still in progress. |
Description | Determining how glucose-dependent insulinotropic polypeptide (GIP) and glucagon-like peptide 1 (GLP1) synergistically regulate beta cell function |
Amount | £289,769 (GBP) |
Funding ID | MR/W000881/1 |
Organisation | Medical Research Council (MRC) |
Sector | Public |
Country | United Kingdom |
Start | 01/2022 |
End | 09/2022 |
Description | ERC CoG, which was ultimately underwitten by the UKRI Horizon Guarantee. |
Amount | € 1,950,000 (EUR) |
Funding ID | EP/X026833/1 |
Organisation | European Research Council (ERC) |
Sector | Public |
Country | Belgium |
Start | 01/2023 |
End | 01/2028 |
Description | GC-globulin as a novel regulator of alpha cell function and glucagon secretion during type 2 diabetes |
Amount | £249,086 (GBP) |
Organisation | Diabetes UK |
Sector | Charity/Non Profit |
Country | United Kingdom |
Start | 06/2023 |
End | 05/2026 |
Title | CRIPSR GLP1R-/- mice |
Description | We used CRISPR to introduce a single nucleotide deletion into exon 2 of the Glp1r. This resulted in a frameshift, nonsense mediated mRNA decay and complete loss of the protein. These mice are useful for pre-clinical testing of drugs targeted against the GLP1R. |
Type Of Material | Model of mechanisms or symptoms - mammalian in vivo |
Year Produced | 2019 |
Provided To Others? | Yes |
Impact | We have provided the mice to labs in the UK and Germany under a standard MTA provision. We are likely to get further requests once the model is published. |
URL | https://www.nature.com/articles/s41467-020-14309-w |
Title | Cell impermeable SNAP tag labels |
Description | SNAP tag labels are usually cell impermeable due to the fluorophore which they use. This precludes use of a number of super-resolution imaging-compatible fluorophores, including SiR, JF549 and JF646. We have synthesised novel SNAP tag labels which incorporate a charged sulfonate, thus rendering any fluorophore cell impermeable. This allows surface proteins to be specifically interrogated. |
Type Of Material | Technology assay or reagent |
Year Produced | 2020 |
Provided To Others? | Yes |
Impact | The novel SNAP tag labels increase resolution of super-resolution microscopy. They also allow the specific interrogation of surface proteins for the first time using nanoscopy approaches. |
URL | https://pubs.rsc.org/en/content/articlelanding/2020/sc/d0sc02794d#!divAbstract |
Title | Chemical probes for the detection and visualization of GIPR |
Description | We developed the GIPR probes and are testing their specificity in pancreatic islets. Our partners (Frank Reimann and Fiona Gribble) are testing the ligands in the brain. We recently filed a European patent to protect intellectual property surrounding these probes, and will look to commercialize going forward. |
Type Of Material | Technology assay or reagent |
Year Produced | 2023 |
Provided To Others? | Yes |
Impact | There is no specific antibody for GIPR, which is holding back understanding of sites of action of this important therapeutic receptor. |
Title | Chemical probes for the detection and visualization of GLP1R |
Description | We extended the color palette of our chemical probes for the detection and visualization of GLP1R. LUXendin645, LUXendin551 and LUXendin762 have been distributed so far to over 25 labs worldwide. https://chemrxiv.org/engage/chemrxiv/article-details/60ff0d5a393cc904c94f1f18 |
Type Of Material | Technology assay or reagent |
Year Produced | 2021 |
Provided To Others? | Yes |
Impact | Antibodies are notoriously unreliable for the detection of GLP1R. This has led to mis-assignment of GLP1R agonist target tissues. For example, some investigators detect GLP1R in the liver, but we now know that there is no GLP1R in the liver. Thus the reported affects of GLP1R agonists on fatty liver disease are likely indirect or via a small immune cell compartment. Our chemical probes specifically and sensitively detect GLP1R. They have been provided to numerous labs worldwide and have increased the accuracy of GLP1R research. Since we no longer have budget or person power, we have most recently entered into a licensing deal with Celtarys Research to supply probe. |
URL | https://www.celtarys.com/categoria-producto/glucagon-like-peptide-1-receptor-glp1r |
Title | Far red antagonist peptide labels for visualising GLP1R |
Description | Far red antagonist peptide labels for visualising GLP1R. The probes are wash free and allow detection of endogenous GLP1R in vitro and in vivo. |
Type Of Material | Technology assay or reagent |
Year Produced | 2018 |
Provided To Others? | Yes |
Impact | We have disseminated probes to multiple labs in the UK, EU and North America. The probes are being used for a number of projects related to GLP1R function, with relevance for the treatment of diabetes. |
URL | https://www.nature.com/articles/s41467-020-14309-w |
Title | SNAP_GLP1R mice for visualization of GLP1R in the living system |
Description | We used CRISPR to knock-in a SNAP self-labelling tag onto the N-terminus of the GLP1R. We can now label GLP1R in vivo in a mammalian species using a range of fluorophores. |
Type Of Material | Model of mechanisms or symptoms - mammalian in vivo |
Year Produced | 2020 |
Provided To Others? | No |
Impact | SNAP_GLP1R mice will help us understand GLP1R distribution and function, as well as test novel drugs. This will impact potentially impact how we treat diabetes, since many drugs target the GLP1R. |
URL | https://pubmed.ncbi.nlm.nih.gov/36653347/ |
Description | Design, synthesis and testing of SNAP labels for visualizing surface proteins |
Organisation | Cornell University |
Department | Weill Cornell Medicine |
Country | United States |
Sector | Academic/University |
PI Contribution | We helped to develop and generate a number of cell-impermeable fluorophores for interrogating surface proteins. We performed trafficking studies using the SNAP_GLP1R. |
Collaborator Contribution | Our partners performed SIMPull experiments, as well as synthesised compounds. |
Impact | The is an inter-disciplinary collaboration. The collaboration has only just started, so outputs are pending. |
Start Year | 2019 |
Description | Design, synthesis and testing of SNAP labels for visualizing surface proteins |
Organisation | Max Planck Society |
Department | Max Planck Institute for Medical Research |
Country | Germany |
Sector | Charity/Non Profit |
PI Contribution | We helped to develop and generate a number of cell-impermeable fluorophores for interrogating surface proteins. We performed trafficking studies using the SNAP_GLP1R. |
Collaborator Contribution | Our partners performed SIMPull experiments, as well as synthesised compounds. |
Impact | The is an inter-disciplinary collaboration. The collaboration has only just started, so outputs are pending. |
Start Year | 2019 |
Description | Design, synthesis and testing of orthosteric peptide labels for visualizing GLP1R |
Organisation | Max Planck Society |
Country | Germany |
Sector | Charity/Non Profit |
PI Contribution | We performed all the pharmacological and biological testing of next generation antagonist far red GLP1R peptide labels. To test specificity of the probes, we also generated GLP1R-/- mice using CRISPR technology. |
Collaborator Contribution | Kai Johnsson's and Johannes Broichhagen's group at the MPI for Medical Research designed and synthesised next generation antagonist far red peptide labels for detecting endogenous GLP1R. They also performed super-resolution microscopy (STED) using Stefan Hell's nanoscopy platform, located in Heidelberg, Germany. |
Impact | We have had numerous requests for probes and animals based upon conference presentations. So far, we have disseminated probes to muliple labs in UK, EU and North American. We have also distributed GLP1R-/- mice under MTA to Imperial College London and Helmholtz Munich, as well as deposited animals into the Mutant Mouse Resource and Research Center. We recently entered into an exclusive licensing deal with Celtarys Research, who commercialize the probes on our behalf, with a profit share agreement. The collaboration is highly multi-disciplinary spanning chemistry, chemical biology, structural biology, physiology and pharmacology. |
Start Year | 2018 |
Description | Development and testing of GIPR probes |
Organisation | University of Cambridge |
Department | Institute of Metabolic Science (IMS) |
Country | United Kingdom |
Sector | Academic/University |
PI Contribution | We have developed GIPR probes. No antibody exists for this GPCR, which is becoming translationally very relevant, since it is targeted by GLP1R/GIPR dual agonists for the treatment of diabetes and obesity. |
Collaborator Contribution | We developed the GIPR probes and are testing their specificity in pancreatic islets. Our partners (Frank Reimann and Fiona Gribble) are testing the ligands in the brain. We recently filed a European patent to protect intellectual property surrounding these probes, and will look to commercialize going forward. |
Impact | The partnership is still in its early phase, but we have already filed a patent, have a mansucript under revision with Eli Lilly and University Cambridge. The collaboration is multi-disciplinary spanning chemical biology, neuroscience, and involves an industrial partner (Eli Lilly). |
Start Year | 2021 |
Description | Endogenous GPCR clustering analysis |
Organisation | University of Birmingham |
Country | United Kingdom |
Sector | Academic/University |
PI Contribution | We collaborated with Prof Dylan Owen's lab to perform dSTORM microscopy, as well as clustering analysis of the ensuing datasets. |
Collaborator Contribution | Dylan's lab provided free time on an ONI nanoimager system (usually 37 GBP per hr), as well as technical assistance with analysis. |
Impact | We recently published a manuscript together in Nature Communications, which shows data from the collaboration. https://pubmed.ncbi.nlm.nih.gov/36653347/ |
Start Year | 2021 |
Description | Purifying mature hESC-derived beta cells using GLP1R probes |
Organisation | University of British Columbia |
Country | Canada |
Sector | Academic/University |
PI Contribution | We developed and validated GLP1R probes, allowing its visualization. |
Collaborator Contribution | The partner generated beta-like cells from hESC, before staining with GLP1R probes and FACS-sorting for assessment of maturity. |
Impact | We have shown that GLP1R can be used to purify the most mature beta-like cells derived from hESC. This is important, since transplants of such cells are in clinical trials, yet performance is still lacking versus native islets. |
Start Year | 2019 |
Description | Understanding crosstalk between androgen and GLP1 signaling |
Organisation | Tulane University |
Country | United States |
Sector | Academic/University |
PI Contribution | We have performed a number of FRET assays to study the effects of androgen on GLP1R signaling in mouse islets and cell lines. |
Collaborator Contribution | They developed the research area and are performing molecular and in vivo studies. |
Impact | We have published a paper in JCI Insight and are currently preparing another manuscript. |
Start Year | 2019 |
Description | Art exhibition to showcase OCDEM research to patients |
Form Of Engagement Activity | Participation in an activity, workshop or similar |
Part Of Official Scheme? | No |
Geographic Reach | Regional |
Primary Audience | Patients, carers and/or patient groups |
Results and Impact | An interactive art show was setup in the atrium of OCDEM, which is also the waiting area for patients referred to tertiary diabetes, lipid and podiatry clinics. |
Year(s) Of Engagement Activity | 2022 |
Description | Presented the labs work to patients with diabetes in Coventry |
Form Of Engagement Activity | A talk or presentation |
Part Of Official Scheme? | No |
Geographic Reach | Regional |
Primary Audience | Patients, carers and/or patient groups |
Results and Impact | I presented the labs work to patients with type 1 and type 2 diabetes as part of a Diabetes UK networking event. |
Year(s) Of Engagement Activity | 2019 |
Description | Vancouver Diabetes Research Day |
Form Of Engagement Activity | A talk or presentation |
Part Of Official Scheme? | No |
Geographic Reach | International |
Primary Audience | Postgraduate students |
Results and Impact | Vancouver Diabetes Research Day for research trainees. I gave one of the Keynote presentations, covering work related to the UKRI awards, as well as mentorship. |
Year(s) Of Engagement Activity | 2020 |
URL | https://diabetesbc.ca/event/vancouver-diabetes-research-day-2020/ |
Description | World Diabetes Day 2020 |
Form Of Engagement Activity | A formal working group, expert panel or dialogue |
Part Of Official Scheme? | No |
Geographic Reach | International |
Primary Audience | Public/other audiences |
Results and Impact | With our Institute Communications Manager, we set up an expert panel for World Diabetes Day 2020. The panel included the Dean of the Medical school, the Research Director for the charity Diabetes UK, clinical academics, basic scientists and patients with lived experience. The virtual panel was streamed live on Zoom and attracted > 500 attendees from all over the world, including patients, general public and students. The format was questions and answers. |
Year(s) Of Engagement Activity | 2020 |
Description | World Diabetes Day 2021 |
Form Of Engagement Activity | Engagement focused website, blog or social media channel |
Part Of Official Scheme? | No |
Geographic Reach | International |
Primary Audience | Public/other audiences |
Results and Impact | I took part in a Q and A with Diabetes UK regarding research funded by Diabetes UK and MRC. The Q and A was hosted on their website as well as Twitter. |
Year(s) Of Engagement Activity | 2021 |
Description | World Diabetes Day 2022 |
Form Of Engagement Activity | Participation in an activity, workshop or similar |
Part Of Official Scheme? | No |
Geographic Reach | Regional |
Primary Audience | Public/other audiences |
Results and Impact | We set up a marquee in town, with interactive displays, as part of World Diabetes Day 2022. |
Year(s) Of Engagement Activity | 2022 |