NIHRDataInsights/HRCSData huggingface dataset:
Title: REGULATION OF HUMAN GLUCOSE HOMEOSTASIS BY THE NOVEL CHC22 CLATHRIN ISOFORM
Funder: Medical Research Council

Abstract: This research addresses how membrane traffic of the GLUT4 glucose transporter in humans is regulated by CHC22 clathrin. Insulin-stimulated delivery of GLUT4 to the cell surface from an intracellular GLUT4 storage compartment (GSC) is the primary mechanism for post-prandial glucose clearance from blood into muscle and fat. We discovered that CHC22 clathrin mediates formation of the human GSC, and accumulates at the GSC in muscle of Type 2 diabetes patients, when GLUT4 is trapped inside due to insulin resistance (IR). CHC22 clathrin is a novel isoform of clathrin that functions in a distinct pathway from the familiar clathrin (CHC17) involved in endocytosis. This research grant will define CHC22 function and regulation with three aims. - Advanced imaging of human muscle cells will produce a comprehensive map of CHC22 membrane traffic within the GLUT4 pathway. The molecular mechanism of changes in CHC22 traffic following insulin treatment and in models of IR in human muscle cells will be addressed through analysis of phosphorylation and changes in CHC22 partner proteins. - Proteomics and functional assays in human myotubes will be used to discover proteins involved in CHC22 regulation under different nutritional conditions. Classical biochemistry will be used to define the mechanism of CHC22 uncoating from membranes and to compare the assembly properties of common genetic variants of CHC22. - CHC22 genotyping will be performed from DNA of human subjects participating in studies of whole-body glucose control (Univ of Bath) to correlate genetic variation with measured parameters of glucose homeostasis. CHC22 transgenic mice will be produced for breeding and nutrition experiments (Univ of Oklahoma) to assess the influence of CHC22 variation on development of IR in mice expressing human GLUT4. These studies will impact the cell biology and diabetes fields by addressing the molecular basis for CHC22 regulation of human glucose metabolism during health and disease


Research activity codes: 1.1 NORMAL BIOLOGICAL DEVELOPMENT AND FUNCTIONING;2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: METABOLIC AND ENDOCRINE
Title: The effect of GLP-1/glucagon 'dual agonists' on amino acid metabolism: potential side effects and solutions
Funder: Medical Research Council

Abstract: Obesity and type 2 diabetes are global pandemics. Novel glucagon-like peptide-1 (GLP-1)/glucagon 'dual agonists' are being developed, combining appetite-suppressive and insulinotropic effects of GLP-1 and glucagon with the energy expenditure (EE) promoting effects of glucagon. However, glucagon induces catabolism of amino acids and excessive glucagon secretion in patients with glucagonoma leads to marked muscle wasting. I have shown that a naturally secreted GLP-1/glucagon dual agonist, oxyntomodulin (OXM), reduces amino acid levels in humans. I have also demonstrated upregulation of hepatic urea cycle enzymes, using a dual agonist analogue. I hypothesise that chronic GLP-1/glucagon dual agonist treatment leads to increased amino acid catabolism and loss of lean body mass. My objectives are to determine: 1. The acute effect of a GLP-1/glucagon dual agonist on amino acid metabolism is glucagon receptor dependent. 2. The chronic effect of a GLP-1/glucagon dual agonist on lean body mass, muscle function, protein metabolism and EE. 3. Whether a high protein diet prevents loss of lean body mass during chronic dual agonist treatment. I will use a long-lasting analogue of OXM, OX-SR, and OX-SR-Glu3 which possesses only GLP-1 receptor action, to: 1. Confirm the effect of a dual agonist on the acute metabolism of amino acids is due to the glucagon component. 2. Determine effects of a dual agonist on lean body mass, muscle function, protein metabolism and EE, when given to animals for 30 days. 3. Determine whether a high-protein diet prevents loss of lean body mass and muscle function when a dual agonist is given to animals for 30 days. 4. Demonstrate a similar effect on loss of lean body mass and amino acid metabolism in humans when OXM is given subcutaneously for 30 days. These findings will establish the safety of GLP-1/glucagon dual agonists prior to clinical use and establish a preventative measure against side effects from these drugs.


Research activity codes: 5.1 PHARMACEUTICALS
Health categories: METABOLIC AND ENDOCRINE
Title: Investigating the role of immature beta cells in insulin release from the intact islet
Funder: Medical Research Council

Abstract: 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.


Research activity codes: 2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: METABOLIC AND ENDOCRINE
Title: High-throughput in vivo profiling of candidate human obesity genes in zebrafish
Funder: Medical Research Council

Abstract: Obesity and associated diseases cause 3.4 million deaths every year. The design and development of new anti-obesity therapies is urgently needed. Exome sequencing studies have identified many new genes that are associated with severe obesity in humans. These genes represent potential new therapeutic targets; however, how they function in obesity is entirely unknown. In this proposal we will utilise a novel, in vivo zebrafish model to identify mechanisms that control human obesity and energy homeostasis. We will use zebrafish for two main reasons: high-throughput in vivo mutagenesis and real-time in vivo imaging. Our preliminary data have identified that Semaphorin 3 signalling regulates obesity susceptibility. First, we will perform a series of experiments to identify how Semaphorin 3 signalling impacts the real-time activity dynamics of neurons within the hypothalamus, which are known to regulate appetite and energy homeostasis. Second, we will implement a new high-throughput automated mutagenesis strategy in zebrafish to comprehensively identify new candidate human obesity genes outside of known obesity pathways. We have established a collaboration with Professor Sadaf Farooqi and the Genetics of Obesity Study to identify candidate human obesity genes, and their function will be assessed in vivo, in zebrafish. Altogether, this proposal will both (i) comprehensively identify new human obesity genes, and (ii) establish how human obesity genes impact dynamic feeding circuit activity with consequences for feeding behaviour and obesity. These studies will be critical for expanding the human obesity gene map, and will provide new molecular targets for anti-obesity therapies.


Research activity codes: 2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: METABOLIC AND ENDOCRINE
Title: NutriPROGRAM: Early-life Nutritional Programming of Metabolic Health through Epigenetic Pathways
Funder: Medical Research Council

Abstract: Pregnancy and infancy are critical periods for nutritional programming of metabolic health. Epigenetic changes seem to have a crucial role in pathways leading from early-life nutrition to metabolic health across the life course. NutriPROGRAM will study key maternal and infant nutrition-related exposures in relation to DNA methylation in mothers and offspring at different ages, and metabolic health outcomes across the life course. It will involve: (1) integrating sets of early-life nutritional factors in mothers and infants; (2) observational and intervention studies across the life course on metabolic outcomes including longitudinal growth and fat development, advanced imaging for body fat, metabolomic profiles and clinical outcomes; (3) cause-and-effect relationships of DNA methylation in the associations of nutrition-related factors and metabolic outcomes across the life course, by advanced causal inference approaches; (4) modification of DNA methylation changes by nutrition, lifestyle, physical activity and microbiota; (5) functional consequences of differences in DNA methylation using RNA expression and tissue specific approaches; and (6) translation into preventive or population-health strategies. NutriPROGRAM will capitalize on rich, existing and complementary nutritional, metabolic and epigenetic datasets from observational and intervention studies in pregnancy, childhood and adulthood, together covering the full life course. It will use knowledge and data infrastructures from different collaborative consortia including the Horizon2020 DynaHEALTH and LifeCycle Projects, the ERA-HDHL ALPHABET project, and the Pregnancy and Childhood Epigenetics (PACE) Consortium. Eventually, NutriPROGRAM will develop a sustainable, translational, international collaboration for current and future studies on nutrition and epigenetics and their role in sustaining good health throughout the lifecourse.


Research activity codes: 2.1 BIOLOGICAL AND ENDOGENOUS FACTORS;3.1 PRIMARY PREVENTION INTERVENTIONS TO MODIFY BEHAVIOURS OR PROMOTE WELL-BEING
Health categories: METABOLIC AND ENDOCRINE
Title: Optimising islet transplantation therapy by targeting the liver niche to promote the long-term engraftment and function of human islets.
Funder: Medical Research Council

Abstract: Islet transplantation consistently restores awareness of hypoglycaemia and stabilises blood glucose control. Unfortunately there is a limited supply of donor pancreases with most recipients requiring 2 donor pancreases. Islets are avascular when transplanted and rely on the process of angiogenesis for engraftment:>60% of islets are destroyed in this avascular state. We have developed particles from the polymer poly(DL-lactide-co-glycolic acid)(PLGA): a biodegradable material used in pharmaceutical formulations in which growth factors can be loaded and their release controlled. The polymer has a galactose moiety attached to it and targets the liver through exploiting the asialoglycoprotein receptor(ASGPR), specific to liver. Keratinocyte growth factor (KGF) is a small polypeptide that binds to the KGF receptor and has proliferative and anti-apoptotic effects on epithelial cells eg. hepatocytes. We have determined the KGF particle dose which in mice promotes proliferation of cells within the liver, increases islet engraftment (histologically) and significantly improves glycaemic control when transplanted with islets via the hepatic portal vein. VEGF particles are also in development. In this proposal the objectives are to: 1. Refine/develop growth factor particles to target liver with appropriate growth factor release kinetics; 2. Demonstrate /confirm efficacy of human islets co-transplanted with these growth factor-particles; 3. Understand the mechanism of action of these growth factors. To answer these objectives we plan to: 1. Refine the release kinetics of KGF from galactosylated KGF-PLGA particles already developed; 2. Further develop VEGF-PLGA particles to target the liver; 3. Assess fate/track particles; 4. Assess short term effects of particles on liver; 5. Assess long-term metabolic effects of particles on islet function; 6. long term negative or positive effect of particles on islet graft. If successful, we will progress to first in man studies.


Research activity codes: 5.2 CELLULAR AND GENE THERAPIES
Health categories: METABOLIC AND ENDOCRINE
Title: The role of chain length in fatty acid uptake in both health and disease: a combined experimental-theoretical modelling approach
Funder: Medical Research Council

Abstract: Metabolic syndrome refers to a range of conditions including obesity, diabetes and high blood pressure, which increases an individual's risk of developing cardiovascular disease. These conditions are typically preceded by elevated levels of circulatory free fatty acids (FFAs), which are taken up by cells, subsequently leading to cellular dysfunction and death. This work will identify the mechanisms by which FFAs cross the plasma membrane (PM) and how FFA size alters the rate and mode of transport. This is important as it is often the longer chained FFAs that exert a toxic phenotype. By identifying such mechanisms, therapies can ultimately be developed to regulate cellular FFA entry, thus providing potential targeted therapies for metabolic syndrome. Raman spectroscopy will be used to image FFAs crossing the PM of cells. These experiments will determine the FFA flux and rate constants, which will then be incorporated into a biophysical model of FFA uptake, based on reaction-diffusion equations. The mathematical model will describe the movement of FFAs both outside and inside the cell, and FFA transport across the PM by both passive diffusion and facilitated transport. By fitting the mathematical model to the data, it will be determined: a) if FFA uptake is facilitated, passive or some combination of the two, b) how FFA size and concentration affect the mode of uptake, and c) how this mechanism is altered in metabolic syndrome. The mathematical modelling predictions will then be validated in a human hepatocyte cell line, a cell that contributes towards insulin resistance, a central characteristic of metabolic syndrome. Hepatocytes will be genetically engineered to over- or under-express components of FFA transport, and then exposed to FFAs of increasing chain length for different durations and at different concentrations. These experimental results will then be used to refine the mathematical model, generating fresh predictions and suggesting further experiments.


Research activity codes: 1.1 NORMAL BIOLOGICAL DEVELOPMENT AND FUNCTIONING
Health categories: METABOLIC AND ENDOCRINE
Title: Introduction of a clinical guideline to manage type-2 diabetes by ayurvedic practitioners in Nepal: intervention development and feasibility study
Funder: Medical Research Council

Abstract: The burden of type 2 diabetes (T2DM) is high in Nepal. Many T2DM patients, especially in rural areas, consult ayurvedic practitioners and use ayurvedic treatments. Ayurveda is a recognised medical system in Nepal, and ayurvedic practitioners are part of the health workforce in Nepal. Despite the clinical effectiveness and safety of several ayurvedic medicines for managing T2DM found in trial settings, strong concerns remain about the sub-optimal T2DM management of many patients in ayurvedic clinical practice. The overarching aim is to explore whether the introduction of a clinical guideline can improve the management of T2DM by ayurvedic practitioners as compared to usual ayurvedic management (i.e., without any clinical guideline). Intending to follow up this work with a future cluster randomised controlled trial (cRCT, the main study), the subsidiary objectives of this early phase study are: 1. To develop the clinical guideline: We will follow the standard clinical guideline development process. The scientific evidence base is not large for many ayurvedic components (e.g., lifestyle) and we will draw analogous evidence from the standard western medicine clinical guidelines. A systematic review will be conducted on the effectiveness and safety of ayurvedic medicines in T2DM management, to generate evidence statements. These statements will be presented to an independent clinical guideline development committee for making the final decisions. Accordingly, the clinical guideline will be drafted and finalised. 2. To determine the feasibility of undertaking the main cRCT: Primary Health Centres (PHCs) with an ayurvedic practitioner will be randomised to intervention or comparator groups. New cases of T2DM, diagnosed by the ayurvedic practitioner, will be recruited. Quantitative data will be collected to estimate essential parameters that are needed to design the main cRCT. Semi-structured interviews will be conducted to explore the study-specific issues.


Research activity codes: 7.3 MANAGEMENT AND DECISION MAKING
Health categories: METABOLIC AND ENDOCRINE
Title: Circadian regulation of liver energy metabolism: translational studies in diabetes and obesity
Funder: Medical Research Council

Abstract: Evidence suggests that circadian control mechanisms in the liver are essential for energy homeostasis, but how bioenergetic substrates communicate with the clock, and how liver circadian metabolism acts on the substrates is unknown. I will model human liver using 3D spheroid culture systems, with primary human hepatocytes (PHH), as immortalized cell lines poorly replicate human liver. These human liver spheroids maintain insulin sensitivity in long term culture and this phenotype can be modulated by different intervention (i.e. glucose/insulin concentration). This technology offers me a perfect research training opportunity to investigate the interplay between energy metabolism, especially lipids, and the hepatic circadian clock, as explained below. The circadian metabolic system in the liver is plastic, and responds to changes in state, such as presence of inflammation. This project capitalises on recent innovations which permit human liver organoids to be used as a translational model for diabetes and metabolic dysfunction. The microlivers will be challenged with lipogenic, high-energy culture medium, to drive lipid accumulation within the hepatocytes. The impact of this challenge, the mechanism underlying human hepatosteatosis, on the core circadian clock will be assessed by tracking the PER2-luc output, and by measuring gene expression profiles. As 90% of the lipogenic programme in the liver lies under circadian control, I will target the core clock components BMAL1 (which stops the clock), and REVERBa, (major link between the clock and lipogenic gene expression). I have in house BMAL1, and REVERBa floxed mice crossed to albumin cre-ERT, which allows post-natal deletion of the clock genes. I will recover the livers, and use these to make microlivers, as above. My primary outcomes will include metabolic gene expression profiles through time, measured using nanostring technology, and this will be supplemented with stable isotope studies.


Research activity codes: 2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: METABOLIC AND ENDOCRINE
Title: Gallstone Disease: A Sentinel Presentation of Cardiometabolic Diseases
Funder: Medical Research Council

Abstract: Cardiovascular disease remains the highest source of mortality in the UK whilst mortality and morbidity from metabolic diseases such as insulin resistance, hypertension and dyslipidaemia are increasing significantly. Identification of patients at risk from cardiometabolic diseases may allow timely intervention with preventative measures. Affecting up to 20% of the population, gallstone disease occurs more frequently in those with risk factors for cardiometabolic diseases such as obesity. Gallstones also share common genetic aetiology with cardiometabolic diseases in the ABCG8/5 cholesterol transporter genes. This project aims to evaluate the relationship between gallstones and cardiometabolic disease and to explore shared genetic aetiology. The project will make use of UK Biobank data, the Edinburgh Type 2 Diabetes Study, Generation Scotland and retrospective data from Information Services Division (NHS Scotland). Data will also be retrieved from NHS Lothian radiology reports using natural language processing to determine gallstone status. The radiological data will be linked to genetic and phenotypic data from the other data sources. The cohort data will be assessed for a temporal relationship between gallstones and cardiometabolic phenotypes using Cox proportional hazards model. A GWAS will be performed to identify single nucleotide polymorphisms (SNPs) associated with development of gallstones. SNPs shared between gallstones and cardiometabolic disease will be assessed for neighbouring genes with plausible mechanistic links. Evidence that gallstones are an intermediate phenotype in the development of cardiometabolic diseases provides a key step in supporting future research into preventative measures in this patient group and will have a key role in personalised, predictive medicine. This research aims to lay the foundation for a trial of aspirin, statins or lifestyle modification in patients with gallstones.


Research activity codes: 2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: METABOLIC AND ENDOCRINE
Title: Development and potential reversal of type 2 diabetes: How critical is vitamin A in the regulation of insulin responsiveness and lipid homeostasis?
Funder: Medical Research Council

Abstract: According to WHO, the global prevalence of diabetes has increased from 4.7% in 1980 to 8.5% in 2014. In the Philippines, the national prevalence in 2008 was already at 7.2%. Although recent clinical and pathophysiological studies have shown that type 2 diabetes is caused by excess, yet reversible, fat accumulation in liver and pancreas, there is convincing evidence that impaired vitamin A signalling and metabolism contributes to this condition. Indeed, serum retinol binding protein (RBP4) correlates with hepatocyte triglyceride concentrations in both animals and humans, and is associated with increased de novo lipogenesis. Binding of retinol-RBP4, but not apo-RBP4, to STRA6 triggers phosphorylation of STRA6, up-regulation of SOCS3 with subsequent suppression of insulin signalling. Retinoic acid receptor (RAR)-alpha signalling is required to maintain glucose-stimulated insulin secretion in pancreatic beta-cells and adequate mitochondrial beta-oxidation in the liver. The project therefore aims to determine how increased RBP4 augments liver de novo lipogenesis via a range of mechanisms, and to determine whether energy restriction breaks this potential vicious cycle. Finally, the project aims to determine whether: i) diet-induced weight loss among diabetics in the Philippines achieves the same pathophysiological changes as in Causcasians in Newcastle; ii) a low energy liquid diet is culturally acceptable; and iii) if substantial weight loss can deliver a clinically superior overall outcome at 6 months compared with management by current best practice guidelines from the Ministry of Health in the Philippines.


Research activity codes: 4.1 DISCOVERY AND PRECLINICAL TESTING OF MARKERS AND TECHNOLOGIES
Health categories: METABOLIC AND ENDOCRINE