NIHRDataInsights/HRCSData huggingface dataset:
Title: The role of novel epigenetic regulators in beta cell development and growth
Funder: Juvenile Diabetes Research Foundation

Abstract: Beta cell replacement therapies hold promise as a bona fide cure for type 1 diabetes. An in depth understanding of the molecular pathways governing pancreas formation and subsequently beta cell differentiation and expansion will be instrumental for achieving this goal. Epigenetic regulatory mechanisms are crucially important for embryonic development, self-renewal and differentiation, yet very little is known concerning how they control the development and growth of beta cells. Preliminary data show that germline knockout mice of the H3K27 demethylases, Utx and Jmjd3, have ventral pancreas agenesis and aberrant islet morphogenesis, respectively. Utx and Jmjd3 are generally known to control developmental programming and cellular growth by promoting differentiation and inhibiting proliferation. This proposal therefore aims to determine the roles of the demethylases in the pancreas. I will first examine Utx- and Jmjd3-expression using antibodies against the demethylases in co-immunofluorescence staining for markers of mature pancreatic cells and their precursors. To dissect essential cell autonomous roles of Utx and Jmjd3 in the pancreas, I will then use the Cre/LoxP system to generate and subsequently analyze conditional mutants with specific deletion of the demethylases in Pdx1+ presumptive pancreatic cells to examine their roles in pancreas morphogenesis, and in the beta cells to evaluate if the H3K27 demethylases control the decline in cell cycle activity associated with ageing. Following Cre-mediated ablation of Utx and/or Jmjd3, Pdx1-Cre;demethylase-/- embryos will be analyzed for defects in early foregut patterning and at later stages for aberrant pancreas and islet morphogenesis, while beta cell specific demethylase knockout mice will be examined for abnormal islet morphogenesis and tested for enhanced beta cell proliferation. Finally, to examine potential redundancy of the demethylases in pancreas development and beta cell growth, I will analyze Utx-flox/Jmjd3-flox double mutant mice by performing developmental stage- and beta cell-specific deletions of both enzymes using available Cre lines. These experiments will yield important knowledge on the roles of Utx and Jmjd3 on specific pancreatic functions which can be readily exploited to control beta cell programming and growth for the treatment of type 1 diabetes.


Research activity codes: 1.1 NORMAL BIOLOGICAL DEVELOPMENT AND FUNCTIONING
Health categories: METABOLIC AND ENDOCRINE
Title: Potential mechanisms of persistent C-peptide in Type 1 diabetes
Funder: Juvenile Diabetes Research Foundation

Abstract: Recent studies, using modern ultra-sensitive C-peptide assays, have challenged the conventional view that patients with Type 1 diabetes (T1D) progress over time to absolute insulin deficiency after the “honeymoon period”. At all durations of T1D, there is a range of endogenous insulin production with variation between individuals, with some individuals having high levels of endogenous despite many years of T1D. Islet destruction in T1D is mediated by pathogenic beta cell specific autoreactive T cells, with T1D occurring due to an imbalance between the pathogenic T cells and islet protective regulatory T cells. This proposal will primarily test the hypothesis that differences between patients endogenous insulin secretion results from differences in their immune phenotype. We have recently assessed stimulated endogenous insulin secretion in 1005 people with long duration T1D around Exeter, UK using the urine C-peptide creatinine ratio (UCPCR). UCPCR correlates well with serum C-peptide in the mixed meal tolerance test. We will identify and study 220 people with log duration (> 5years) T1D selected from the top and bottom extremes (sextiles) of endogenous insulin secretion adjusted for duration (110 C-peptide high and 110 C-peptide low). We will confirm C-peptide status of participants using a mixed meal tolerance test and at the same time take blood for analysis of peripheral blood mononuclear cells (PBMCs), for analysis of autoreactive T-cells . We will use established, state of the art, investigations in London, UK and Leiden, Holland to assess islet specific and regulatory T cell responses from the cryopreserved PBMCs from these patients. This will include measurement of frequency and phenotype of CD8+ autoreactive T cells by Quantum Dot peptide-MHC-multimers, measurement of utoreactive T-cells by dye dilution technology and assessment of regulatory T cell frequency and function using multiparameter flow cytometry and an in vitro co culture suppression assay. In addition we will assess differences in the presence of islet autoantibodies (GAD, IA2 and ZnT8), genetic predisposition and historical glycaemic control. If we identify a difference in the immune phenotype of patients with persistent C-peptide in longstanding type 1 diabetes, this will focus efforts on preserving C-peptide in Type 1 diabetes towards achieving this in more patients. Any difference we see would provide important information about what pathways to focus future immune treatments on. Conversely a lack of association would suggest that other factors such as genetic programming of beta cell mass, tight initial glycemic control, or possibly the ability of beta cells to regenerate, are more important in preservation of beta cell function.


Research activity codes: 2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: METABOLIC AND ENDOCRINE
Title: Small-molecule agonists of GLP-1R for the treatment of Type-2 diabetes
Funder: Innovate UK

Abstract: Type-2 diabetes has assumed almost epidemic proportions, resulting in a significant financial burden to healthcare systems in developed countries. GLP-1R agonism represents one of the mainstays of therapy for type-2 diabetes, but existing GLP-1 mimetics necessitate injection and have been associated with severe side effects. Through the use of its proprietary NMR-based approach to drug design, C4X Discovery (C4X) has built the first comprehensive pharmacophore GLP-1R agonist 'map' and used this to derive the smallest orthosteric agonist of GLP-1R so far reported. As part of the project, C4X will explore whether this molecule can form the basis of a chemical optimisation programme that will enable the creation of orally-available and cheaper alternatives to exisiting therapies, thereby providing a significant healthcare benefit in terms of increased patient convenience, improved compliance and reduced economic burden.


Research activity codes: 5.1 PHARMACEUTICALS
Health categories: METABOLIC AND ENDOCRINE
Title: Rejuvenating beta cell antigen-specific regulatory T-cells for adoptive therapy
Funder: Juvenile Diabetes Research Foundation

Abstract: We aim to develop a new immune-based strategy for cure/prevention. Our approach combines two key elements. (i) Basic science studies in our lab have shown the existence of a population of T cells that regulate immune responses whenever they encounter beta cells (antigen-specific Tregs). (ii) Laboratory investigations in our lab and others show the feasibility of taking adult cells and re-programing and rejuvenating them for use as therapeutic agents (induced pluripotent stem cell; iPSC technology). We aim to combine these two elements and make rejuvenated, long-lived, antigen-specific Tregs that we can grow in abundance. If these proof-of-concept studies yield decisive results, these Treg-iPSCs could be grown in the laboratory and injected back into subjects with, or at risk of Type 1 diabetes, as a means to control the immune destruction of beta cells.The rationale for this approach is that animal studies have shown that infusion of beta cell-specific regulatory immune cells can reverse autoimmune diabetes, even in late disease models. There are technical difficulties associated with isolation and expansion of these cells to sufficient number. We propose a rejuvenation approach for antigen-specific Tregs, based on iPSC technology. iPSCs can be generated from virtually any cell type of the human body by temporarily expressing specific pluripotency factors. iPSCs display embryonic stem cell-like features and can be coaxed to differentiate into many different cell types, including immune cells. Recent studies have shown that the generation of iPSCs from clonally expanded antigen-specific effector T cells and subsequent redifferentiation into T cells relieves their aged phenotype while maintaining their original antigen specificity. Here we propose to investigate whether this rejuvenation strategy can be applied to a previously generated cohort of beta cell antigen-specific T cell clones, which have regulatory properties and appear strongly linked to Type 1 diabetes pathogenesis. This proposal is unique in that it brings together the expertise of stem cell biologists (Henckaerts and collaborators) and Type 1 diabetes immunologists (Tree and Peakman) co-located within the same research program at Kings College. We are the only lab to have generated and characterized beta cell antigen-specific human Treg clones. . If we can demonstrate that these clones can be rejuvenated and expanded without affecting their specific immune cell characteristics and normal chromosomal makeup, we would be able to establish a preclinical and clinical development program that could lead to first-in-man studies of a therapy that promises safe modification of the immune system in Type 1 diabetes patients.


Research activity codes: 5.2 CELLULAR AND GENE THERAPIES
Health categories: METABOLIC AND ENDOCRINE
Title: Understanding T cell migration via inflamed endothelium in Diabetes Type 1
Funder: Juvenile Diabetes Research Foundation

Abstract: Type 1 diabetes (T1D) is caused by immune cells called T cells attacking and damaging the insulin producing beta cells in the pancreas. This leads to a fall in the production of insulin and to the patient having to take insulin injections for the rest of their lives. Whilst this insulin therapy does work, it does not work very well. Patients (often young children) have to finger-prick to check their blood glucose values, and to inject insulin upto 5 times a day. The disease is associated with risk of low blood glucose as well as with risk of blindness, kidney failure and premature death.We believe we have identified an agent that can prevent T1D. It works by preventing T cells (which normally circulate in the blood) from entering and attacking the insulin producing beta cells. We have called this agent PEPITEM for peptide inhibitor of trans endothelial migration. In our experiments so far, we show in a test tube that PEPITEM prevents T cells from crossing blood vessel walls. We have gone on to show in an animal model and it does the same. We now wish to move on to see if PEPITEM can prevent T1D. In order to do so, we will inject PEPITEM into a mouse model of T1D called the NOD (non obese diabetic) mouse. This is a common model of T1D that has been used in a number of experiments over many years. We believe it is a good first step in testing the ability of PEPITEM to prevent disease. If it works, we will move on to testing it in other animal models and well as doing the important safety and dosing work that is required before it can be given to patients. These latter experiments are outside the scope of this current application.


Research activity codes: 5.1 PHARMACEUTICALS
Health categories: METABOLIC AND ENDOCRINE
Title: The role of GPRC6A agonists in satiety
Funder: Biotechnology and Biological Sciences Research Council

Abstract: Recent research has highlighted the importance of nutrient sensing in the regulation of food intake and metabolism. However, the mechanisms by which a high protein diet regulates food intake are largely unknown. We have found that specific amino acids which agonise the G-protein coupled receptor family C group 6 member A (GPRC6A) reduce food intake when peripherally administered, and that this effect may be mediated via the gastric hormone ghrelin. We now aim to determine the molecular mechanisms behind these effects and whether they represent physiological systems controlling food intake and body weight. Aim To investigate the mechanisms by which GPRC6A agonists reduce food intake Objectives 1) To determine the site at which GPRC6A agonists act to cause their anorectic effects. 2) To investigate the mechanisms mediating the effects of GPRC6A agonists on food intake. 3) To determine the physiological role of GPRC6A agonists in the regulation of satiety. Together this programme of work will determine the mechanisms by which the GPRC6A receptor regulates food intake and suppresses ghrelin levels, and investigate the physiological role of GPRC6A in the regulation of energy homeostasis. The role of GPRC6A in food intake has not previously been investigated.


Research activity codes: 1.1 NORMAL BIOLOGICAL DEVELOPMENT AND FUNCTIONING
Health categories: METABOLIC AND ENDOCRINE
Title: Investigation of role of hypothalamic glucokinase in the control of food intake
Funder: Biotechnology and Biological Sciences Research Council

Abstract: Within the CNS glucokinase (GK) is expressed at high levels in the hypothalamus, notably the paraventricular nucleus (PVN), ventromedial nucleus (VMN) and arcuate nucleus. We have conducted pilot experiments to investigate the physiological role of GK in the hypothalamus. We hypothesised GK may play a role in the regulation of food intake. This is supported by our finding that fasting increased GK activity specifically in the arcuate. To investigate the physiological importance of this change we injected AAV expressing GK (AAV-GK) into the arcuate nucleus this produced an increase in GK activity similar to that seen following a fast. This increase in GK activity resulted in a decrease in food intake. This effect on food intake may be due to an effect on glucose appetite as rats with increased arcuate GK had reduced intake of glucose compared to controls, whilst intake of fructose was unaffected. This decrease in glucose intake suggests GK may be the mechanism underlying the glucostatic regulation of food intake. This is supported by our finding a preload of glucose reduced food intake in rats with increased arcuate GK compared to controls. We will now investigate the physiological role of GK in the control of long term food intake. We will do this by injecting AAV encoding GK or anti-sense GK into the arcuate of groups of adult rats. Their food intake and body weight will be measured. The effect of addition of glucose to the diet on food intake and body weight will be investigated. This will be performed using both normal and high fat diet. This will investigate whether GK has a physiological role in the glucostatic regulation of food intake. Possible mechanisms by which GK influences food intake will be investigated. Together this data will examine the physiological role of GK in the regulation of energy homeostasis. It will establish the utility of hypothalamic GK as a pharmaceutical target.


Research activity codes: 1.1 NORMAL BIOLOGICAL DEVELOPMENT AND FUNCTIONING
Health categories: METABOLIC AND ENDOCRINE
Title: GPR103 has multi-tissue effects on health and metabolism
Funder: Biotechnology and Biological Sciences Research Council

Abstract: Few factors act in a coordinated manner on both central and peripheral metabolic pathways to produce anabolic effects in the body. Recent evidence demonstrates that the peptide, QRFP, has major actions in the brain and key metabolic tissues, such as the pancreas and adipose, to increase energy intake, reduce glucose tolerance and cause lipogenesis. We will demonstrate that different populations of QRFP receptors, GPR103A and GPR103B, mediate its varied effects in mice with the two receptors knocked out. We will generate a mouse in which the Qrfp gene promoter drives the expression of cre recombinase and the GFP reporter. This will allow us to record the electrical activity of QRFP neurones following different stimuli. Using the latest adeno-associated virus technology, we will also express selectively in QRFP neurones an anterograde tracer to determine QRFP neuronal projections or "designer" receptors. The latter will allow us to activate or inhibit selective populations of QRFP neurone in vivo by administering a "designer" drug intraperitoneally. Finally, we will generate a loxSTOPlox-Qrfp mouse, which is effectively a peptide null mutant and which is expected to express adverse metabolic phenotypes. By crossing this mouse will other cre-expressing mice, e.g. nestin-Cre or adiponectin-Cre, we can genetically re-introduce QRFP to selective tissues in order to induce different gains in function. This work will validate QRFP as a key regulator of systemic metabolic processes, and potentially as a valuable and novel pharmacological target for the treatment of metabolic disease.


Research activity codes: 1.1 NORMAL BIOLOGICAL DEVELOPMENT AND FUNCTIONING
Health categories: METABOLIC AND ENDOCRINE
Title: Biological resonance: matching internal timing to environmental fluctuations
Funder: Biotechnology and Biological Sciences Research Council

Abstract: This proposal examines the impact of physiological timing on energy balance in mammals. Our studies will determine the degree to which metabolic homeostasis is compromised when feeding and activity patterns oppose our internal clock, a common feature of modern society. We will test this hypothesis by modulating environmental input (light and food) into the clockwork, by weakening internal coordination of different body clocks, and by genetically altering clock speed in a tissue selective way to drive internal de-synchrony. As we disrupt the alignment of the internal circadian clockwork with environmental cycles, and progressively drive internal desynchrony (between liver and SCN), mice will undergo comprehensive longitudinal assessment of behavioural (locomotor activity) and physiological rhythms (Tb, metabolic gas echange) and determination of metabolic efficiency based upon gross anatomical measures (BW, adiposity), nutrient profiling in the liver (TG, glycogen content) and serum (inc. glucose, TG, FFA, cholesterol, insulin, glucagon), and glucose tolerance testing. Genetic manipulations will employ established and validated mouse lines (Ck1etau, Albcre), as well as a new SCN-selective cre-driver line recently created by our collaborator Michael Hastings (LMB Cambridge). Critically, we employ and further refine a novel technology to longitudinally monitor liver gene expression in vivo in free moving mice. This will allow for the first time, assessment of transcriptional changes in response to altered feeding schedules in real time. By this means, we will identify how feeding cycles drive not only the local hepatic clock-work genes (ie BMAL1), but crucially we will measure its impact on specific hepatic metabolic-regulating genes. To our knowledge, this is the first study to use resonance protocols and genetic alteration of clock speed to determine the true impact on circadian timing on mammalian physiology.


Research activity codes: 1.1 NORMAL BIOLOGICAL DEVELOPMENT AND FUNCTIONING
Health categories: METABOLIC AND ENDOCRINE
Title: Impact of food processing on the blood cholesterol-lowering effect of cereal beta-glucan
Funder: Biotechnology and Biological Sciences Research Council

Abstract: The effect of beta-glucan (BG), a water-soluble form of fibre, in lowering LDL cholesterol concentrations has been demonstrated by many research groups. This beneficial impact has been accepted as a valid health claim for BG, in non-processed or minimally processed forms of oats and barley, by EFSA. The mechanism(s) of action of BG are strongly linked to its capacity to generate high viscosity in the gastrointestinal tract and a subsequent decrease in bile reabsorption through decreases in rates of digestion, inhibition of lipolysis and/or transport of mixed micelles. The BG may also directly interact with lipase, bile salts and the mucus layer, leading to decreased bile recycling. However, the precise mechanisms are still not fully understood. Moreover, the cholesterol-lowering activity of BG varies significantly depending on food type, processing and storage conditions, mainly due to depolymerisation and poor solubility of BG. The main aims of this project therefore are to understand (a) the mechanism(s) by which BG lowers blood cholesterol and (b) how the processing of BG-containing foods attenuates this capacity of the polymer. A multidisciplinary approach involving a range of in vitro methods, biochemical (e.g. in vitro digestion, enzyme kinetics) and biophysical (e.g. rheology, fluorescence microscopy), will be used. Samples of purified BG and BG-containing cereals and real foods (porridge & biscuits) will be characterised (e.g. molecular weight, polymer dissolution), before and after food processing, and their behaviour during in vitro digestion and their interactions with BS and lipase(s), will be studied. These samples will be subjected to a range of sophisticated in vitro digestion models allowing viscosity and interactions to be determined during breakdown and transport. This work will provide new information on how BG can be processed and added to food matrices in a way that maintains optimal health effects.


Research activity codes: 3.3 NUTRITION AND CHEMOPREVENTION
Health categories: METABOLIC AND ENDOCRINE
Title: Using crop genetics to understand the importance of dietary resistant starches for maintaining healthy glucose homeostasis
Funder: Biotechnology and Biological Sciences Research Council

Abstract: Our aim is to develop a systematic basis for increasing the intake of resistant starch in the diet in order to protect the function of insulin-secreting pancreatic beta-cells and improve blood glucose homeostasis in an ageing population. Age related declines in beta-cell function and glucose homeostasis increase susceptibility to type 2 diabetes, the incidence of which has doubled in the UK since 1996. We and others have shown that carbohydrates such as resistant starch that escape digestion in the small intestine lead to improved insulin sensitivity and first phase insulin response. We propose that short chain fatty acids liberated by colonic carbohydrate fermentation causes receptor-mediated improvements in beta-cell function. In this project we will define which starches and food matrices are most effective in protecting beta-cell function and discover the basis of this protection. The results will underpin development of a roadmap for increasing the most effective types of resistant starch in the diet via a concerted approach involving crop breeding and food processing. Our project utilises a suite of genetically-defined natural mutants of peas, representing two types of resistant starch in a variety of genetic backgrounds. Experiments with static and dynamic digestion models will examine starch digestibility and colonic fermentability in raw and processed materials and purified starch. Selected samples will be physically characterised to establish which starch/matrix features determine digestibility and fermentability, which will be confirmed in human studies. In parallel we will carry out human trials on selected peas to assess effectiveness in beta-cell protection. Volunteers aged 55 - 65 will be used to assess physiological functions related to susceptibility to diabetes including measures of insulin release, gut hormones, colonic microbiota and the microbiota activity.


Research activity codes: 3.3 NUTRITION AND CHEMOPREVENTION
Health categories: METABOLIC AND ENDOCRINE