NIHRDataInsights/HRCSData huggingface dataset:
Title: Exosomes - circulating cardioprotective nanoparticles
Funder: Medical Research Council

Abstract: Exosomes are nanometre-sized, lipid membrane vesicles that ferry proteins and other components through the circulation and appear to have a signalling role. Exosomes circulate in the blood of healthy individuals, but their properties and purpose are virtually unknown. In pilot data we have isolated exosomes from healthy rat plasma and shown that they are cardioprotective in a Langendorff isolated, perfused, rat heart system. Our objective is to characterize these exosomes, to determine their mechanism of cardioprotection, and to evaluate the potential of circulating exosomes as an autologous therapeutic agent. This will be achieved using established techniques for exosome purification and analysis which require the use of a preparative ultracentrifuge and Nanosight equipment for determining particle size distribution and concentration. The Langendorff isolated heart preparation will be used extensively for evaluating cardioprotection, and confirmed using our established in vivo rat MI model. A recovery model will be utilized establish the importance of this protection on a longer time-scale relevant for patients. Protein content of plasma exosomes will be determined using proteomics and Western blotting. This will inform design of experimental protocols to identify the mechanism of cardioprotection. Molecular characterization of exosomal interaction and/or fusion with the heart will be achieved using multiphoton microscopy of exosomes labelled with fluorescent nanodots. We have already identified one candidate cardioprotective protein (HSP70) and its role will be investigated using a HSP70-neutralizing antibody developed by Prof Multhoff (Munich). Investigation of the cardioprotective component of exosomes may lead to identification of novel cardioprotective pathways or proteins. We stand ready to fully exploit other lead proteins and cardioprotective pathways identified, in terms of potential patents, publications and further experiments.


Research activity codes: 2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: CARDIOVASCULAR
Title: Investigation of MiR-140-5p in Humans and Experimental Models of Pulmonary Arterial Hypertension.
Funder: Medical Research Council

Abstract: Pulmonary arterial hypertension (PAH) is a devastating and life threatening condition for which there is no pharmacological cure. Dissecting the molecular mechanisms that drive pathogenesis is crucial for the development of effective treatments. Several studies have highlighted the importance the Bone Morphogenetic Protein Receptor Type II (BMPR2) and related signalling pathways in PAH, but how these cause the disease remains unclear. MicroRNA, a recently described class of RNA, are proposed to play a key role in regulating health and disease by modulating the translation of target mRNA. In-house data that I have performed demonstrate that whole blood microRNA-140-5p (miR-140-5p) is decreased in patients with idiopathic and scleroderma associated PAH. In Silico analysis predicts that miR-140-5p targets molecules downstream of BMPR2. I therefore hypothesise that miR-140-5p expression is decreased in PAH, resulting in reduced BMPR2 signalling and contributing to disease pathogenesis; even in the absence of a BMPR2 mutation. To test this hypothesis I will use a combination of clinical samples, in vitro cell culture and rodent models of PAH to validate the initial microRNA (miR) array results, and subsequently quantify changes in the key predicted targets of miR-140-5p in pulmonary artery endothelial cells (PAEC) and pulmonary artery smooth muscle cells (PASMC). We will measure the effect of miR-140-5p on PASMC and PAEC phenotype, before determining the effect of exogenous miR-140-5p on disease phenotype in rodent models. This data will provide clear and novel information on the function of miR-140-5p in pulmonary vascular cells and the temporal expression in two animal models of PAH. This will allow direct translation to our clinical data showing reduced miR-140-5p in PAH, and establish whether reduced miR-140-5p levels are causative, or a bystander in PAH pathogenesis. Finally this will provide insight into the utility of miR-140-5p levels as a biomarker in PAH.


Research activity codes: 4.1 DISCOVERY AND PRECLINICAL TESTING OF MARKERS AND TECHNOLOGIES;2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: CARDIOVASCULAR
Title: TRANSLATING HUMAN EMBRYONIC STEM CELL-DERIVED ENDOTHELIAL CELL THERAPY TO THE CLINIC
Funder: Medical Research Council

Abstract: Human embryonic stem (hES) cells and induced pluripotent stem (iPS) cells hold broad potential in regenerative medicine. Such stem cells can generate all cell types upon stimulation to differentiate along defined cell commitment pathways. We are interested in the mechanisms that govern stimulation of these stem cells into vascular ECs and their subsequent application to regenerative medicine. We have recently published studies that show the potential of human ECs derived from hES cells (Kane et al., ATVB, 2010; Stem Cells., 2012). We showed a very efficient production of ECs in just 10 days following initiation of differentiation. This rapid system induced robust changes, i.e. very effective reduction in pluripotency markers and induction of endothelial selective markers. We also defined the "therapeutic" potential of these cells in cell cultures and in vivo by demonstrating the induction of angiogenesis and blood flow improvement in a mouse model of limb ischaemia. These results suggest that these cells and this differentiation system may be useful clinically in the treatment of ischaemic conditions in patients with cardiovascular disease. This present application builds upon pilot data showing that we can generate human endothelial cells from hES cell lines under GMP-compliant conditions (achieved through TSB funding). We now wish to develop this translational agenda towards a first-in-man trial through careful assessment of these cells. This will be achieved by final protocol optimisation and validation, safety studies, efficacy and biodistribution studies. This will be conducted by bringing together key researchers to work together towards this goal.


Research activity codes: 5.2 CELLULAR AND GENE THERAPIES
Health categories: CARDIOVASCULAR
Title: Randomised controlled trial of podoconiosis treatment in northern Ethiopia.
Funder: Medical Research Council

Abstract: The objective of this trial is to test the effectiveness of lymphoedema management in podoconiosis patients. We plan to achieve this using a pragmatic, single-blind, randomised controlled trial (RCT) with three intervention 'arms' that will compare two models of podoconiosis lymphoedema management in the community with delayed treatment (as control). 'Standard' podoconiosis lymphoedema intervention consists of twice-weekly group meetings with instruction and practical demonstration of foot hygiene by a community project assistant. Management comprises: foot hygiene, use of emollient, bandaging, exercises to improve lymph drainage, elevation of the foot of the bed and use of shoes and socks. 'Intensive' podoconiosis lymphoedema management consists of 'standard' plus additional 5 times per week clinic-based manual lymphatic drainage for one hour for the first two weeks. 900 adult podoconiosis patients located in independent households will be randomised, 300 to each arm. The primary outcome is incidence of acute dermatoadenolymhangitis (ADLA, a common, debilitating complication of untreated podoconiosis). Secondary outcomes include adherence with treatment, clinical stage of disease, lower leg and foot circumference, quality of life, prevalence of entry lesions, duration of ADLA, perceived stigma and economic productivity. The principal analyses will be conducted at 6 and 12 months of follow-up, and will be based on the rate of ADLA events using multivariable Poisson regression. Results will be disseminated rapidly through the National Podoconiosis Action Network in Ethiopia, and through Footwork, the International Podoconiosis Initiative, to stakeholders in other podoconiosis-endemic countries, as well as through more conventional academic channels such as peer-reviewed articles. The demand for information from potential podoconiosis care-giving organisations is high, and application of results is likely to be swift.


Research activity codes: 7.1 INDIVIDUAL CARE NEEDS;6.8 COMPLEMENTARY
Health categories: CARDIOVASCULAR
Title: Investigation of novel protein pathways which regulate vascular smooth muscle cell phenotype in neointimal hyperplasia
Funder: Medical Research Council

Abstract: Despite recent advances in stenting, neointimal hyperplasia from increased vascular smooth muscle (VSM) cell proliferation remains a significant clinical problem, particularly in peripheral arterial disease. Delineation of the mechanisms which change VSM cells from a quiescent to a proliferative cell phenotype during neointimal formation is still urgently required to alleviate this health burden. Our recent study has uncovered a protein, uncharacterised in the cardiovascular system, which we revealed has an important role in regulating VSM cell gene expression. This protein is called phosphoprotein enriched in astrocytes-15 (PEA-15). We have already determined that PEA-15 binds the mitogen-activated protein kinases (ERK1/2) and acts as a cytoplasmic "tether" preventing ERK1/2 activating gene expression in the nucleus. ERK1/2-induced gene expression is directly associated with growth factor-mediated proliferation and, by preventing this, PEA-15 maintains VSM cells in a quiescent state. Our preliminary evidence now indicates that PEA-15 expression is substantially decreased in the VSM cells of arteries from a mouse model of neointimal hyperplasia. In the current proposal we will address the hypothesis that PEA-15 expression is decreased in neointimal hyperplasia and directly contributes to VSM cell proliferation during neointimal development. Both in vivo and in vitro models will be used to fully uncover the effects and mechanisms which modulate PEA-15 expression in neointimal formation. We will further examine the therapeutic potential of increasing PEA-15 expression to reduce ERK1/2-induced gene expression and inhibit neointimal formation. Our pilot data has indicated that vitamin D can increase PEA-15 expression and this will be examined in models of vascular disease. In this project, we will therefore uncover a novel mechanism responsible for the development of neointimal hyperplasia and, in addition, reveal a viable therapeutic approach.


Research activity codes: 2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: CARDIOVASCULAR
Title: Developing new mature, functional vascular networks in ischemic disease
Funder: Medical Research Council

Abstract: The proposal consists of two parts. Part 1 will determine the balance of factors required to provide a physiologically relevant neovasculature in a relatively sparsely perfused tissue. This will be carried out using the rat mesenteric microvascular angiogenesis assay, using adenoviruses expressing VEGF, Ang1, eNOS, netrin, ephrins and DLL4. Angiogenesis, arteriolargenesis (formation of novel arterioles from existing capillaries), and arteriogenesis (expansion of the arterial system from existing arterioles) will be measured using intravital microscopy and confocal imaging of the mesentery. Flow, permeability coefficients (solute permeability, hydraulic conductivity, oncotic reflection coefficient), compliance, and vasoreactivity will be determined. Endothelial intracellular calcium in response to agonists and physical forces in vivo will be measured using intravital dyes perfused into the vessels. This will dissect mechanisms through which vascular remodelling occurs under physiological conditions. Part 2 will transfer this knowledge, built on existing expertise, into animal models of human cardiovascular disease. It will develop and validate novel vectors for the delivery of key remodelling agents to cardiac and skeletal muscle cells. We will use adenovirus (AdV), validated for delivery of genes to these cells, and generate novel viruses expressing Ang1, eNOS and VEGF under control of tissue specific promoters, inducible by hypoxic conditions. These will then be validated in vivo, and applied to disease models, as adenoviruses injected locally. The effect of these viruses on muscle blood flow and cardiac function will be assessed to determine whether the approach of developing normal vasculature is more efficient at recovering animals from cardiovascular insults than angiogenic or arteriogenic agents alone.


Research activity codes: 2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: CARDIOVASCULAR
Title: The role of autophagy during cardiac forward and reverse remodelling
Funder: Medical Research Council

Abstract: We will examine the role of autophagy in reverse remodelling and the role of TSC1/2 to determine the size and nature of the heart in mTORC1 signalling. In this project, we will mainly use two different genetically modified animals, Atg5- and TSC2-deficient mice. Both are cardiac-specific deletion models generated by crossing floxed mice with cardiac-specific Cre transgenic mice. To confirm the involvement of mRNA translation and autophagy, the TSC2-deficient animals will be crossed with 4E-BP1 and Atg5-deficient animals, respectively. We will use three different stress models: i.e., pressure overload by means of transverse aortic constriction and angiotensin II infusion using by osmotic mini-pumps. LV function will be determined by echocardiography and hemodynamic measurement. The extent of hypertrophy will be examined by physiological measurement, histological analysis, and mRNA expression of marker genes using quantitative RT-PCR. Apoptosis is estimated by TUNEL staining. The level of necrosis will be evaluated by nuclear labelling with ligation of pfu generated probes. Intracellular signalling pathway will be examined by immunoblotting using specific antibodies and kinase assay. At in vitro level, we shall isolate cardiomyocytes from 8 week-old mice and incubate them with H2O2 or isoproterenol to confirm the effect of gene deletion on cell death. Viability of the cells will be measured by Cell Titre Bleu assays. We will examine the signal transduction mechanisms in isolated cardiomyocytes from rat neonatal or mouse adult hearts using immunoblotting. Cardiomyocyte hypertrophy will be estimated by measurement of cell surface area and immunohistochemical analysis. Cardiomyocyte contractile function will be measured using edge detection and Ca2+-sensitive fluorescent dye. Protein translation activity will be evaluated by polysome assay. Autophagy levels will be examined by Western blot analysis and ultra-structural analysis.


Research activity codes: 2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: CARDIOVASCULAR
Title: Role of defective Kv7.4 channels in vascular disease.
Funder: Medical Research Council

Abstract: Opening of voltage-dependent potassium channels encoded by the KCNQ4 gene (Kv7.4) maintain vascular smooth muscle in a relaxed state and also underlie vasodilations by physiological mediators. However, dysfunction of Kv7.4 is a major feature of arteries from hypertensive animals. More importantly, as Kv7.4 has a key role in renal arteries then defective Kv7 channels may be central to the development of high blood pressure. The intended project will build upon the foundation studies by the applicant and will determine by quantitative PCR and western blot analysis how the expression of KCNQ genes alters in the renal and cerebral arteries as rats develop hypertension. KCNE1-5 gene products alter the biophysical properties of Kv7.4 so this project will use a molecular interference approach to assess the role of KCNE-products in vascular tone and will determine how the expression of these genes is affected by hypertension. These studies will be consolidated by isometric tension recording and patch clamp electrophysiology on isolated smooth muscle cells and HEK293 cells heterologously expressing KCNQ4, which will define how the Kv7.4 channel is stimulated by beta-adrenoceptor agonists. The factors dictating the membrane abundance of Kv7.4 and the impact of angiotensin II on protein levels will be elucidated by Western blot analysis and immunocytochemistry.


Research activity codes: 2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: CARDIOVASCULAR
Title: Integrated cardiopulmonary modelling for the investigation of the management of disturbed tissue perfusion
Funder: Medical Research Council

Abstract: We will design and develop novel computational models of the human cardiac and vascular systems and integrate these with our published, validated pulmonary models. The resulting multi-organ modelling will be multiscalar and will have great scope and depth of detail (e.g. a branching three-dimensional vessel-bed containing variable depths of cells, tissue perfusion distribution affected by endogenous mediators and drugs, heart comprising four chambers with stretch- and perfusion-affected myocardial contractility and valvular outlets, heart contained within a pericardium/mediastinum/thorax complex that will be externally compressible to allow simulation of cardiac massage). Each compartment will affect neighbouring compartments and flow between compartments will be calculated arithmetically for micro-epochs (<1 ms) and a new dynamic state calculated by moving compartmental contents as determined by calculated flows. We will apply novel "smart" validation techniques to test the veracity and fitness-for-purpose of the integrated models; these will include robustness analysis techniques, prospective validation against single-patient data streams and replication of previously published clinical research. Using these integrated multi-organ models we will investigate the disturbed physiology of low cardiac output states and potential therapeutic strategies. Such "shock states" are poorly understood and are a common feature of critical illness, which consumes a large part of the healthcare budget. Initially, we will address poorly understood aspects of such states (e.g. How does manipulation of blood viscosity affect cellular oxygenation during shock? How does permissive hypercapnia affect cellular oxygenation and pH during shock?). Using the knowledge generated in this phase, we will test algorithms for the management of various shock states in large populations of in-silico subjects.


Research activity codes: 6.9 RESOURCES AND INFRASTRUCTURE (TREATMENT EVALUATION);2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: CARDIOVASCULAR
Title: MICA: Reducing cardiovascular disease risk through replacement of saturated fat in milk and dairy products
Funder: Medical Research Council

Abstract: UK exceeds dietary targets for SFA recommended for CVD risk reduction. Milk/milk products are one of the largest sources (30% of total SFA). Recent data show likely benefits of increased milk consumption in reduced CVD risk thus consuming less milk/products to lower SFA intake may be counterproductive. Recent studies by our group (funded by BBSRC DRINC) show the potential of dairy cow nutrition to produce reduced SFA and increased cis-MUFA milk in a sustainable fashion. The hypothesis that consumption of such low-SFA dairy foods will reduce CVD risk in those at increased risk, compared with normal dairy foods, via mechanisms to increase secretion of endothelial-derived vasodilators will be tested. Initially, reduced SFA/enhanced cis-MUFA milk, cheese and butter will be produced by cow dietary changes. These modified foods will be used in double-blind, randomised, controlled, crossover chronic and acute studies with adults at increased CVD risk (n=54) to assess their effects in the background diet (12-weeks) and in test meals on biomarkers of CVD. These will include traditional (lipids and blood pressure) and novel markers (vascular function, determined by flow mediated dilatation and stiffness, pulse wave velocity), postprandial lipids, biomarkers of inflammation and vascular function (cellular adhesion molecules, endothelial progenitor cells). Furthermore, this project will use a co-culture system (human aortic smooth muscle cells overlayered with endothelial cells) which more closely mimics physiological systems, to examine the responses to plasma isolated during the acute postprandial studies on gene expression and protein levels of biomarkers of vascular function and endothelial vasodilators (including nitric oxide), providing a novel insight into mechanisms of action of low-SFA dairy products. These studies will provide unique evidence to inform public health policy on sustainable food-based dietary recommendation for CVD risk reduction.


Research activity codes: 4.1 DISCOVERY AND PRECLINICAL TESTING OF MARKERS AND TECHNOLOGIES;3.3 NUTRITION AND CHEMOPREVENTION
Health categories: CARDIOVASCULAR
Title: PRedicting Out of OFfice Blood Pressure in the clinic (PROOF-BP)
Funder: Medical Research Council

Abstract: Background Accurate blood pressure (BP) measurement is important to ensure antihypertensive treatment is targeted appropriately. New guidelines recommend the use of the 'gold standard' out of office measurement to confirm all diagnoses of hypertension. Our own pilot work suggests that the characteristics of BP measured repeatedly in the clinic can be used to predict out office BP thus, reducing the need for expensive out of office measurement. This method of assessing clinic BP characteristics has only been evaluated in patients with uncontrolled hypertension. Further work is needed to translate this novel method into routine clinical practice. Aims 1)Establish the predictive abilities of multiple clinic BP measurements for out of office BP in a general population. 2)Develop a 'triaging' tool which reduces the number of patients with suspected hypertension being referred for out of office BP measurement. 3)Pilot a clinical trial assessing general practitioner (GP) use of this triaging tool for patients with suspected and existing hypertension. Methods Using a dataset of approximately 1,400 patients (>18 years; white, South Asian or Black ethnicity from within the UK and Canada), BP characteristics from multiple clinic measurements will be estimated using polynomial regression modelling. These will be compared to out of office BP monitoring (ambulatory and home) and the predictive abilities of clinic BP for white coat and masked hypertension will be evaluated. Using the resulting diagnostic thresholds for these conditions, a web based tool for use in clinical practice will be developed. This will be piloted in GP practices in Birmingham, providing preliminary data to inform a full clinical trial. Scientific/Medical opportunities This work presents an opportunity to improve the diagnosis of hypertension and accuracy of routine clinic BP measurement. This could lead to significant cost savings and improved quality of care for patients with hypertension.


Research activity codes: 6.3 MEDICAL DEVICES
Health categories: CARDIOVASCULAR