NIHRDataInsights/HRCSData huggingface dataset:
Title: RAMP2 and the glucagon receptor: important in the pathogenesis of non-alcoholic fatty liver disease?
Funder: Medical Research Council

Abstract: Evidence suggests that reduced hepatic glucagon action may contribute to development of non-alcoholic fatty liver disease (NAFLD), a condition that affects 1 in 3 British adults. Receptor activity-modifying protein-2 (RAMP2) interacts with the glucagon receptor (GCGR) and affects its ligand binding and intracellular signaling. To date this interaction has not been studied in hepatocytes. My pilot data demonstrates that RAMP2 upregulation decreases activity of glucagon in Huh7 hepatoma cells. Further, I have shown that RAMP2 expression is dramatically increased in a mouse model of NAFLD. Additionally, in Huh7 cells upregulation of RAMP2 occurs in response to palmitate exposure, a fatty acid increased in the circulation of patients with metabolic syndrome. This data supports my theory that upregulation of RAMP2, via its effects on the glucagon receptor, is important in the pathogenesis of NAFLD. My experiments will establish the effect of upregulated hepatic RAMP2 on lipid metabolism in mice and determine if increased hepatic RAMP2 expression contributes to dietary-induced steatosis. I will use recombinant adeno-associated virus to specifically upregulate RAMP2 in hepatocytes of adult mice. Furthermore, I will employ histological techniques to establish if the association between RAMP2 and steatosis holds in humans. Alongside, I will investigate the cellular basis of the effects of RAMP2 on the glucagon receptor. This will include use of an established protocol to investigate trafficking of the GCGR using a SNAP-tag in conjunction with confocal microscopy and flow cytometry. This project will dramatically enhance understanding of the pathogenesis of a very common and serious medical condition, with the potential for therapeutic exploitation. More generally, this project will expand current understanding of receptor-interacting proteins, with far-reaching applications for science in related and diverse fields.


Research activity codes: 1.1 NORMAL BIOLOGICAL DEVELOPMENT AND FUNCTIONING;2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: ORAL AND GASTROINTESTINAL
Title: The role of HMGB1 in liver injury and repair - mechanisms and therapeutic interventionsUniversity of Edinburgh
Funder: Medical Research Council

Abstract: Liver disease is a major medical problem. Activation of inflammation serves to promote tissue repair but can contribute to injury exacerbation. Understanding mechanisms whereby key danger signals act as master regulators that drive pathological events or promote injury resolution is critical for developing novel therapies and biomarkers of disease status. Recognition of endogenous molecules as danger signals released by cells upon infection or injury is an emerging concept in Hepatology. My previous research has discovered that one such molecule, High Mobility Group Box-1 (HMGB1), is a biomarker of acute and chronic liver injury and key driver linking cell death to sterile inflammation. Extracellular HMGB1 function is controlled by post-translational modifications. However, its precise regulatory role in liver disease has not been investigated, nor has its therapeutic potential been fully elucidated. This fellowship seeks to dissect the mechanistic role of HMGB1 in the progression and restoration of liver injury and to test novel strategies for its pharmacological modulation. I will test the hypothesis that during liver injury, defined cell types increase the expression of, post-translationally modify and secrete specific HMGB1 isoforms. These isoforms then act as regulatory autocrine and paracrine alarm signals that trigger and/or sustain necrosis and inflammation. To achieve this goal I will utilise novel, cell specific, conditional knock out and transduction models in a loss and gain of function approach. These findings will be reinforced with targeted studies investigating mechanisms by which HMGB1 isoforms regulate parenchymal/non-parenchymal cell cross-talk. Finally, I will determine the utility of anti-HMGB1 biologic therapy in clinically relevant models of liver disease. I will optimise its use through measuring circulating HMGB1 itself and by a novel, real time and non-invasive, HMGB1 hepatic expression reporter in a combined and stratified approach.


Research activity codes: 2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: ORAL AND GASTROINTESTINAL
Title: Interrogating ITGA11 signalling in rodent and human for targeted anti-fibrotic therapy in liver fibrosis.
Funder: Medical Research Council

Abstract: Fibrosis of the liver is characterised by progressive accumulation of ECM proteins and is a major cause of morbidity and mortality. End-stage liver fibrosis can be treated by liver transplantation, but this is limited by donor numbers. Anti-fibrotic therapies are an urgent priority, but no drugs have been approved to date. Pathological ECM and scarring drive progression of fibrosis. Discovering how to block scar production and the signalling in surrounding cells that perpetuates fibrosis represents a very attractive therapeutic avenue. Cells contact the scar via cell surface integrin (ITG) receptors and signal intracellularly to mediate a broad range of cellular responses. We have identified ITGA11, and its partner subunit ITGB1, drives and perpetuates major pro-fibrotic signals in liver fibrosis. Moreover, intracellularly, downstream pathways involving P21-activated kinase (PAK) can be targeted pharmacologically to improve fibrosis in rodent models of liver fibrosis. The pressing challenges are now to test these data in vivo to determine the function and response of ITGA11 during liver fibrosis and if loss of ITGA11 prevents or ameliorates liver fibrosis. Moreover, we need determine the genome-wide effects of ITGA11 signalling to help identify new downstream pathways that may facilitate novel therapeutic intervention and interrogate the role of PAK to determine whether drugs already in development can be used as antifibrotic therapy in liver fibrosis. To facilitate the translational capability of this work, importantly, all pathways will be verified directly in human biopsy tissue from patients with varying severity of fibrosis and molecularly in primary human culture models of fibrosis. Taken together, these experiments are anticipated to prove an important role for ITGA11 in organ fibrosis and assist in identifying new pathways of value in the search for new therapies.


Research activity codes: 2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: ORAL AND GASTROINTESTINAL
Title: Alpha-1 antitrypsin to improve the efficacy of hepatocyte transplantation in children with liver-based metabolic disease
Funder: Medical Research Council

Abstract: Hepatocyte transplantation (HT) is an alternative treatment to liver transplantation for certain inborn errors of metabolism, to date demonstrating some success, but not fulfilling its long-term promise. One of the major hurdles to sustained success of this therapy is the early cell loss of ~70% of hepatocytes following their infusion. This is largely due to the instant blood-mediated inflammatory reaction (IBMIR), with activation of complement and coagulation pathways. Effective inhibition of IBMIR should significantly improve hepatocyte engraftment and function. Alpha-1 antritrypsin (AAT) is an endogenous protease with anti-inflammatory, anti-thrombotic and anti-apoptotic effects. AAT significantly improves efficacy of islet transplantation in models of diabetes. A cynomolgous model of diabetes showed islet function for more than 700 days in those treated with AAT (versus 90 days in controls). As there are several parallels between islet transplantation and HT we hypothesize that a similar effect will be seen in HT. We have shown in vitro that AAT is significantly anti-thrombotic and anti-inflammatory in a Chandler loop containing human hepatocytes and whole blood. Metabolic activity was improved in hepatocytes cultured in AAT. The proposed project will test proof of principle that AAT use in hepatocyte transplantation improves engraftment and function of cells in small animal models. Following on from this a phase IIa non-randomised open-label clinical trial will study AAT in prevention of the IBMIR in children undergoing hepatocyte transplantation for two forms of liver-based metabolic disease: Crigler-Najjar syndrome and Urea Cycle Defects. The primary outcome measure will be of efficacy (demonstrable recovery of enzyme activity) at 24 weeks post transplant which will be measured using disease-specific markers. A proteomic approach will develop biomarkers of cell survival and function and explore the mode of action of AAT in animal models and clinical trial.


Research activity codes: 5.1 PHARMACEUTICALS;6.1 PHARMACEUTICALS
Health categories: ORAL AND GASTROINTESTINAL
Title: Pathogenesis of Ulcerative colitis: The role of BRINP3 in colonic epithelial barrier function and mucosal inflammation
Funder: Medical Research Council

Abstract: Background: Ulcerative colitis (UC) is a chronic inflammatory bowel disease (IBD) affecting 146000 individuals in the UK. It reduces quality of life, increases morbidities and risk of developing colorectal cancer placing significant burden on healthcare. The exact mechanism of this disease is still unknown. Complex interaction between genetic predisposition, environment factors and intestinal microbiota are believed to play role in causing immune dysregulation within the mucosa. Recent work in our laboratory has shown that there is a down-regulation of BRINP3 in colonic mucosal of UC patients compared to healthy controls. This finding was irrespective of the presence or degree of mucosal inflammation indicating that low expression of BRINP3 is unique to UC and may be playing an important role in pathogenesis. We have also shown that the low expression of BRINP3 in the mucosa is caused by increased methylation. The role of BRINP3 in humans is currently unknown. Aims: 1. Determine the molecular and physiological role of BRINP3 in bowel mucosa and how it affects the function of the bowel mucosa and immune response in mice 2. Characterise the gene regulation and role of BRINP3 in colonic epithelial cells Methodology: The changes in cell proliferation, permeability and inflammation of the colonic mucosa will be examined using BRINP3 knockout mice and well characterised models of experimental colitis. The regulation of BRINP3 gene expression will be studied at the mRNA and epigenetic level. I will investigate the protein complexes and regulatory processes involving BRINP3 in colonic epithelial cells through the generation of reporter constructs, biochemistry and confocal microscopy. Scientific and medical opportunity: If low BRINP3 expression is shown to be playing a role in UC, an agent that increase its expression could potentially be developed as a novel treatment.


Research activity codes: 2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: ORAL AND GASTROINTESTINAL
Title: Enteric nervous system progenitor cells from aganglionic gut for Hirschsprung's therapies
Funder: Medical Research Council

Abstract: The aim of this project is to determine mechanisms that regulate the behaviour of progenitor cells from the aganglionic gut of HSCR patients. Objectives: 1.To determine the expression and role of key molecules of the Notch signalling pathway in aganglionic HSCR gut and in derived ENSPC. 2.Characterise and compare differentiation and proliferation responses to Notch, 5-HT and EDN3 signalling in ENSPC derived from aganglionic HSCR gut with those from ganglionic gut. 3.To analyse the effect of Notch, 5-HT and EDN3 signalling on restoring functionality in aganglionic gut explants ex vivo. Methods: -Collection and isolation of cells from human ganglionic and aganglionic colon samples during surgical procedures (HSCR patients and controls). -Culture of ENSPCs as neurospheres in primary cultures. -Lentiviral labelling of neurosphere cells for constitutive expression of GFP to enable cell tracking. -Analysis of components of the Notch signalling pathway in ENSPC prior to isolation and during culture, by observation of expression and subcellular localization of Notch receptors and activation-signalling intermediary RBPkJ, using immunohistology with confocal microscopy. -Assessment of the roles of: (a)Notch signalling - using chemical inhibition (DAPT) and siRNA knockdown of RBPkJ. (b)5-HT signalling - using pharmacological 5-HT4 receptor agonists, e.g. Mosapride citrate or Prucalopride and antagonists, e.g. SB-207266 or GR113808. (c)EDNRB /EDN3 signalling - using EDN3 and pharmacological EDNRB antagonists e.g. BQ-788 or A192621. -Assessment of neuronal differentiation and functionality in gut explants using inverted phase contrast microscopy to assess contractility and Diamtrak software to measure frequency and amplitude of contractions after transplantation of aganglionic gut derived neurospheres into cultured aganglionic embryonic bowel explants with or without prior manipulation of neuronal differentiation using the siRNA and pharmacological approaches.


Research activity codes: 2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: ORAL AND GASTROINTESTINAL
Title: Biominerals in Health and Disease
Funder: Medical Research Council

Abstract: Biomedical research is dominated almost exclusively by interests in organic molecules. However, we and others are showing increasing evidence for large inorganic structures (typically nanominerals) being involved in healthy cell communication and signalling and not just normal storage and structure or ectopic pathology, for which they are well known. Our mission is to identify mineral structures and their cellular interactions in biology to further our understanding of human health and disease and, where appropriate, to exploit lessons learned through the engineering of novel mineral analogues that may offer fresh treatment modalities in human disease. We have a clear focus on nanoparticulate mineral structures, especially in terms of their occurrence and handling in the gastrointestinal tract. Over the next four years' of research we intend to (i) better describe why there are endogenous nanomineral pathways in the gastrointestinal tract and how these facilitate cell sampling of luminal macromolecules, (ii) demonstrate to what extent engineered nanoparticles, to which we are increasingly exposed, hijack these pathways and are systemically absorbed in humans, (iii) delineate whether dysfunction in the cellular handling of endogenous or exogenous nanomineral underlies the cause of Crohn's disease and (iv) show whether engineered nanoparticles can be exploited to establish selective cell activation and thus have immunotherapeutic properties (e.g. for cancer treatments).


Research activity codes: 2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: ORAL AND GASTROINTESTINAL
Title: MECHANISM OF INFLAMMATION IN ENVIRONMENTAL ENTERIC DYSFUNCTION
Funder: Medical Research Council

Abstract: BACKGROUND: Malnutrition is responsible for 45% of global child deaths, and via long-term effects on health and development it perpetuates poverty across societies and through generations. Chronic malnutrition (stunting) fails to respond to improvement in diet and may be driven by Environmental Enteric Dysfunction (EED), a highly-prevalent, sub-clinical inflammatory enteropathy. The mechanism of inflammation in EED is undefined. OBJECTIVES: 1. To characterise intestinal epithelial cell (IEC)-dependent barrier (dys)function in a murine model of EED. 2. To test the hypothesis that endogenous inflammatory signals from lamina propria leukocytes (LPL) are induced by bacterial translocation and exacerbate IEC-dependent barrier dysfunction, thereby establishing a vicious cycle in EED. 3. To identify endogenous inflammatory pathways and mediators contributing to IEC-dependent barrier dysfunction in EED and to prove the concept that interruption of these pathways improves mucosal immune homeostasis in EED. 4. To assess applicability of findings from the murine model of EED to human populations by extending the IEC monolayer system to investigate consequences of signalling by LPL isolated from patients with EED on IEC-dependent barrier function. METHODOLOGY: Murine work will centre around a model of 'EED' driven by provision of a moderately protein- and lipid deficiency diet with induction of dysbiosis. The same conditions will be applied to strains with cell type-specific modulation of microbial recognition and response pathways. Complementary work with IEC monolayers will allow elucidation of the endogenous pathways and mediators responsible for impeding IEC-dependent barrier function OPPORTUNITIES: The results will help to generate adjunctive immunomodulatory approaches that could be subjected to clinical trials. By targeting redundant inflammatory activation I aim to develop strategies to help support healthy growth of children living in poverty.


Research activity codes: 2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: ORAL AND GASTROINTESTINAL
Title: Fungal suppression by human gut Vdelta2 T-cells in mucosal immunity and inflammation
Funder: Medical Research Council

Abstract: Background: Gut dysbiosis is now recognized as a major driver of the rapid increase in many chronic diseases over the last century, but the influence of commensal fungi on human tissue inflammation is largely unknown. The applicant, Dr Neil McCarthy recently observed that human gut tissue can be protected against fungal outgrowth by using the bacterial metabolite HMB-PP to activate Vdelta2 T-cells (which are absent in rodents). Since host responses to gut fungi are strongly implicated in inflammatory bowel disease (IBD), we will determine whether this novel mechanism of anti-fungal immunity in human mucosal tissue is disrupted in IBD. Hypothesis: Bacterial activation of Vdelta2 T-cells restricts fungus growth and protects against human gut inflammation. Method: Human blood and healthy/inflamed gut tissue will be treated with antibiotics to ablate bacteria and promote fungal growth (either added Candida or endogenous fungi) with/without Vdelta2 T-cell activation by HMB-PP. Key mediators of fungal suppression will be identified using blocking antibodies, recombinant cytokines, and therapeutic reagents that modify Vdelta2 T-cell function in vivo. We will also assess the fungal-protective effects of biofilm formation and genetic deficiencies in the BTN3A proteins that regulate Vdelta2 T-cell responses (patients identified by East London Genes & Health project). Fungal isolates will be identified by MALDI-TOF. Outputs: We will determine how Vdelta2 T-cells suppress intestinal fungi, the range of fungal species restricted by this mechanism, and how gut inflammation and IBD therapies interact with this process. Assays in patients with BTN3A mutations will generate crucial new data on how these proteins regulate Vdelta2 T-cell function and inform the design of novel therapies. Several Vdelta2-targeted drugs have already been developed that potently activate these cells in vivo and could potentially be used to increase patient protection against fungi and treat IBD.


Research activity codes: 2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: ORAL AND GASTROINTESTINAL
Title: Management of chronic hepatitis B in Africa: is a one-stop assessment of liver disease enough? The MATCH-B study
Funder: Medical Research Council

Abstract: Infection with hepatitis B virus (HBV) is highly endemic in Africa and is a leading cause of death mainly from cirrhosis and hepatocellular carcinoma. The World Health Organization has recently called for HBV elimination and scaling up screen-and-treat interventions in particular in sub-Saharan Africa. Between 2011-2014 the PROLIFICA project-the first "screen-and-treat" intervention for HBV in Africa-has demonstrated that the vast majority (90%) of HBV-infected adults in west Africa are inactive carriers (IC) and do not require antiviral therapy. Whether these subjects are at risk of liver disease progression and need regular follow-up is unknown in the African settings. The aim of this study is 1) to determine the rate of HBV reactivation and liver disease progression in HBV inactive carriers in Africa and 2) the efficacy of antiviral therapy to reduce HBV-related complications in the African context. We will re-assess the untreated (n=1,825) and treated patients (n=283) previously enrolled in the PROLIFICA project. Using reference tests (e.g HBV DNA and Fibroscan) patients will be re-assessed after a minimum of 4 years following the first liver assessment. In the untreated cohort (mainly IC) the primary objective will be to determine whether patients who did not meet treatment eligibility criteria at baseline remain so without progression of liver disease. In the treatment cohort the primary objective will be to assess the clinical effectiveness of tenofovir in West African patients. If untreated patients do not exhibit significant liver progression it may be possible to limit the number of follow-up of the millions of patients with inactive hepatitis B in Africa where healthcare resources are seriously limited. This study will provide unique longitudinal data on chronic hepatitis B in Africa and will therefore contribute to the development of national and regional strategies for the control of HBV infection in Africa adapted to the local environment.


Research activity codes: 2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: ORAL AND GASTROINTESTINAL
Title: Human gastruloids: an in vitro system for the study of human gastrulation
Funder: Medical Research Council

Abstract: Over the last few years, together with our collaborator Matthias Lutolf at EPFL (Lausanne, Switzerland), we have developed a robust ESC based 3D model of mouse embryogenesis in culture. In our system, small and defined numbers of ESCs undergo symmetry breaking, germ layer specification and gastrulation like movements in a manner that mimics the embryo. These processes happen with spatial and temporal patterns that parallel the embryo and are both robust and reproducible. The resulting organoids, that we call 'gastruloids', develop the primordial of ectodermal, mesodermal and endodermal derivatives with a remarkable axial organization. We feel that we are now in a position to extrapolate this system to human ESCs and have obtained preliminary results suggesting that this is indeed feasible. Our background work with mouse ESCs and the preliminary results with human ESCs form the basis for this project in which we propose to ask 1. What are the molecular events that underlie the establishment and elaboration of the embryonic axes in humans. 2. The sequence of cellular and molecular events underlying the process of gastrulation. 3. How do early developmental events in human and mouse embryos compare with each other and what are the relative roles that signal and transcription factor networks play in these differences. The project will deliver a unique, robust, reproducible and ethically safe system for the study of post implantation human development, gastrulation and the specification of the primordial for tissues and organs. The experimental system will open up significant opportunities in the areas of disease modelling, iPSC validation, drug testing and, fundamentally, knowledge about human biology which, at the moment is inaccessible.


Research activity codes: 1.1 NORMAL BIOLOGICAL DEVELOPMENT AND FUNCTIONING
Health categories: ORAL AND GASTROINTESTINAL