NIHRDataInsights/HRCSData huggingface dataset:
Title: Predicting acute rejection following kidney transplantation by microarray analysis of the T cell transcriptome
Funder: Medical Research Council

Abstract: Acute rejection remains a significant problem post-renal transplantation. Even a single acute rejection episode has a strong negative correlation with long-term graft outcome. Currently there is no method of predicting acute rejection before it becomes clinically apparent. Microarray analysis has led to the development of biomarkers predictive of clinical outcome in a number of settings. We are proposing to use microarray analysis of purified leucocyte subsets to look prospectively for signatures which can predict acute rejection. We have recently shown that patients with autoimmune disease can be subdivided on the basis of CD8 T cell gene expression into two groups with very different clinical prognoses (see Appendix 1). Interestingly, the same subgroups exist in normal controls suggesting they may play a more general role in response to infection, vaccination and possibly transplantation. Given the pivotal role T cells have as mediators of transplant rejection, we plan to determine if similar subgroups exist in transplant recipients and whether they predict acute rejection. Blood samples will be obtained from transplant recipients - one immediately prior to transplantation (D0) and one 3 months post-transplant (D90). Blood will be subject to FACS analysis to measure leucocyte subset populations and the remaining blood will be processed to isolate separate cell populations for microarray analysis. Throughout the study, a comprehensive record of clinical data will be documented. The following analyses will be performed to address the aims of the study: 1. Microarray expression data from T cell samples will be subjected to supervised hierarchical clustering using the genes that best define subgroups of patients in autoimmune disease to determine whether similar subgroups exist in transplant recipients. Subgroups identified by this analysis will be analysed to look for correlations with acute rejection (AR) and other clinical parameters. 2. Cell subset data will be subjected to unsupervised analysis to identify transcriptional changes that might then be associated with AR. Genes identified will be further examined for functional significance. 3. Gene expression changes associated with End-Stage Renal Failure and contributing to the increased incidence of infection in this population will be sought. The scientific opportunities of this research include the combined application of techniques rigorously optimised within the host laboratory together with bioinformatics approaches to identify cell-specific gene expression changes predictive of acute rejection. The medical application of such a discovery would be more individualised therapy post-transplantation.


Research activity codes: 4.1 DISCOVERY AND PRECLINICAL TESTING OF MARKERS AND TECHNOLOGIES
Health categories: RENAL AND UROGENITAL
Title: The importance of Spleen tyrosine kinase (Syk) as a potential therapeutic target in glomerulonephritis
Funder: Medical Research Council

Abstract: Antibody-mediated glomerulonephritis (GN) is a major cause of end stage renal disease. Current treatments for GN are non-specific, not always effective, and associated with significant toxicity. More effective treatment is needed. We have shown that R788, an oral prodrug of R407, a small molecule inhibitor of spleen tyrosine kinase (Syk) is remarkably effective in both preventing and treating established disease in an antibody-dependent animal model of crescentic GN that is characterized by early macrophage infiltration into glomeruli (nephrotoxic nephritis, NTN). Syk is an intracellular tyrosine kinase that has an established role in immunoreceptor signaling. There is also accumulating evidence that Syk is involved in post-receptor signaling of various cytokines including TNFa, IFNa and IL-1a. I wish to establish the precise role of Syk in NTN and the mechanisms by which Syk inhibition acts, with a view to developing a rationale for inhibiting Syk in the treatment of human GN. The proposed study incorporates a detailed in vitro analysis of the effects of Syk inhibition on cultured rat and human cell lines (macrophages and mesangial cells) and nephritic rat glomeruli ex vivo, using a variety of techniques including standard and cell-based ELISA, western blot and microsphere cytokine assays. siRNA techniques will be employed to determine the specificity of R406 in inhibiting Syk dependent pathways. I will also examine the effect of Syk inhibition on autoantibody production in vivo, using an animal model of glomerulonephritis that relies on antibody production following immunization with the a3 chain of type IV collagen (experimental autoimmune glomerulonephritis). The proposal also includes a study of human material. The presence of activated Syk and its downstream targets will be assessed in biopsy specimens from patients with GN using immunohistochemistry (IHC), allowing me to identify those patients in whom Syk inhibition should be a therapeutic target. I will determine the pattern of urinary and tissue cytokine expression in these patients, using microsphere cytokine assays of urine samples and real time PCR of tissue obtained from biopsies by laser capture microdissection. By correlation with the IHC findings, I may be able to identify surrogate markers of Syk activation in tissue or urine that could be used clinically to identify patients who would benefit from Syk inhibition. I expect that the results of these studies should clarify the potential role of Syk in human GN, and may form the basis of future clinical trials using Syk inhibition as a therapeutic strategy.


Research activity codes: 5.1 PHARMACEUTICALS;2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: RENAL AND UROGENITAL
Title: Development of an efficient, robust MS-based platform for early detection of acute kidney injury
Funder: Medical Research Council

Abstract: Acute kidney injury (AKI) is a frequent and serious condition in critically-ill patients, increasing mortality rate independent of other risk factors. Currently, serum creatinine is monitored to detect AKI; this is an inadequate test, enabling only late-stage detection. Earlier AKI detection is an important component of reducing renal morbidity, and mortality in the critical care population. New biomarkers cystatin C, kidney injury molecule-1(KIM-1), neutrophil gelatinase-associated lipocalin (NGAL) and interleukin-18 (IL-18), promised earlier diagnosis, but results were not convincing when validated in independent studies. Proteome analysis, by capillary electrophoresis mass spectrometry (CE-MS), has enabled the identification and validation of a novel urinary biomarker panel for AKI detection. However, CE-MS is slow, expensive, and inappropriate for routine intensive care unit (ICU) use. This proposal will establish an alternative analytical platform to assess our novel biomarker panel, which is efficient, inexpensive and acceptable for routine ICU use. Two platforms - matrix assisted laser desorption/ionisation-time of flight (MALDI-TOF) and multiple reaction monitoring (MRM) triple-quad-MS - will be explored as dual development strategy to optimise outcome.


Research activity codes: 4.2 EVALUATION OF MARKERS AND TECHNOLOGIES
Health categories: RENAL AND UROGENITAL
Title: Evaluating and targeting C5a receptor in pyelonephritis
Funder: Medical Research Council

Abstract: Urinary tract infection (UTI) caused by uropathogenic E. coli is one of the most common diseases that have the characteristics of frequent recurrence or persistence of infection. In a severe form, it can result in renal failure, chronic kidney disease and death from overwhelming sepsis. Complement, an integral part of the innate immune system, has been described as a double-edged sword since it defends the host against infection, but can also cause harm when activated in an uncontrolled manner. Our previous studies have shown that not only are most human uropathogenic E. coli strains resistant to complement mediated killing, but that complement deficiency in fact protected mice from the development of renal infection. This suggests that the role of complement in UTI is an overwhelmingly damaging one. Given that C5a, a potent inflammatory peptide generated by complement activation, has been implicated in the pathogenesis of various inflammatory and infectious diseases, together with our preliminary data, we hypothesise that C5a is a significant pathogenic mediator in UTI. The proposed research will investigate if and how C5a contributes to the pathogenesis of UTI and explore the therapeutic potential for preventing and treating resistant UTI by targeting C5a receptor (C5aR). To accomplish this, we will employ a murine model of ascending UTI induced by human uropathogenic E. coli, C5aR knockout (C5aR-/-) mice, bone narrow (BM)-chimeras (where the mice have C5aR only on renal parenchymal cells or on BM-derived migratory cells), C5aR ligand (C5a), C5aR antagonist (C5aRa), purified neutrophils and primary renal tubular epithelial cells in our studies. Using these tools, we intend to: i) determine if the absence of C5aR signalling in receptor-deficient or ?blocked mice protects from acute pyelonephritis (APN) and bladder inflammation; ii) investigate the impact of C5aR signalling on epithelial and leukocyte dysfunction and its influence on APN; iii) to build a therapeutic strategy by evaluating the effect of C5aRa given via the urethra into the bladder. We envisage that this study will define a previously unrecognised pathogenic role for C5a in renal infection and provide the mechanistic information about this role, namely that excessive C5a generated during UTI, through acting on C5aR in renal parenchymal cells and infiltrating neutrophils, contribute to tissue destruction, bacterial entry and impairment of resolving bacterial infection. We also see this proposal providing essential background information to pursue novel therapeutic approaches to prevent and treat UTI.


Research activity codes: 5.1 PHARMACEUTICALS;2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: RENAL AND UROGENITAL
Title: Development Clinical Studies - investigation into the efficacy of Mirococept in renal transplantation
Funder: Medical Research Council

Abstract: We are seeking to define a therapeutic approach to the prevention of damage to the kidney during the period immediately after transplantation. This damage is linked to activation of the complement system which occurs through recognition of the grafted organ as ?non-self? as shown in a MRC funded programme of work. We have approached this problem firstly by engineering a recombinant form of a human complement regulatory molecule known to be present in the normal kidney and secondly by modifying this molecule so that it can be delivered to the graft by perfusion before transplantation and retained within the kidney during the period that damage is thought to occur. Preclinical and exploratory clinical studies have indicated that this approach is safe and has the potential to reduce the incidence of delayed graft function in renal transplantation. We are now seeking to conduct a larger-scale multi-centre Phase II study to investigate the clinical utility of the approach. This will at the same time validate therapeutic targets identified by animal work.


Research activity codes: 6.4 SURGERY;6.1 PHARMACEUTICALS
Health categories: RENAL AND UROGENITAL
Title: Evaluation of recombinant complement factor H as therapy for orphan renal diseases Dense Deposit Disease & atypical HUS.
Funder: Medical Research Council

Abstract: Complement factor H (FH) is a 155kDa human plasma glycoprotein (500-800mg/L). Its major activities are limiting the amplification of the Alternative Pathway (AP) in both the fluid phase and on self surfaces. Mutations in FH are associated with the kidney diseases Dense Deposit Disease (DDD) and atypical Haemolytic Uraemic Syndrome (aHUS), The common polymorphic variants V62 and H402 are strongly associated with development of age related macular degeneration (AMD). Factor H replacement is a possible treatment strategy for such diseases. Plasma therapy has been effective in some cases of aHUS associated with heterozygous FH mutations. Secondly, combined liver-kidney transplantation appears to be an effective means of preventing recurrence of aHUS in renal transplants among individuals with renal failure due to FH-associated aHUS. Murine data shows that plasma purified murine and human FH can control C3 turnover in short-term studies in the only mouse model of DDD, the Factor H-deficient mouse strain (CFH-/-). The inability to generate large-scale quantities of murine FH and the rapid development of an immune response to human FH prevented long-term administration of both these agents. Critically therefore, the efficacy of FH in preventing or ameliorating DDD in the CFH-/- mouse remains unexplored. Furthermore, the effects of exogenous FH in the only mouse model of aHUS (CFH-/- transgenic for a mutant mouse FH protein that functionally mimics human aHUS-associated FH mutations, CFH-/-.FH 16-20) also remain unknown. My supervisors have developed and applied for a patent for a method making FH protein using the yeast Pichia pastoris. The human protein is structurally and functionally fully active. The murine FH (rmFH) protein will be manufactured, purified and biophysically characterised using SDS-PAGE, dynamic light scattering and mass spectrometry. The binding affinity of rmFH for heparin, C3b and iC3b will be assessed by surface plasmon resonance (SPR) in comparison to plasma-derived murine FH. To assess complement regulatory functions of rmFH, AP cofactor assays on SDS-PAGE and Decay Acceleration Activity (DAA) measured by SPR will be performed. The rmFH protein will be used to perform in vitro studies examining rmFH binding to murine renal endothelium and tissue sections and to assess efficacy in extended studies in relevant animal models of DDD and aHUS. This work will advance the project to the point of being able to develop the product for human testing in renal disease and subsequently to evaluate FH as a therapy for the commonest cause of late-onset blindness AMD.


Research activity codes: 5.1 PHARMACEUTICALS
Health categories: RENAL AND UROGENITAL
Title: Manipulation of Vascular Endothelial Growth Factor isoform-repertoire as a potential therapy in chronic kidney disease
Funder: Medical Research Council

Abstract: Deranged glomerular permeability and subsequent proteinuria are cardinal features of glomerular disease which is identified(2008 Renal Registry report) as the primary cause of End Stage Renal Failure(ESRF) in 56.5% of cases, including those with presumed glomerular disease without histological characterisation and those with diabetic glomerular damage (24%). In turn ESRF is associated with increased morbidity (Na/water retention, hypertension, dyslipidaemia, vascular disease) and reduced longevity (5 year survival on dialysis = 46.3% [equivalent Grade II carcinoma stomach] and 25.1% if diabetic). Furthermore, the presence of protein in the urine is a strong predictor of cardiovascular risk in the general population, even in those individuals who have high normo-albuminuric levels. This project will attempt to define the molecular basis for deranged glomerular permeability using a unique combination of transgenic animals, functional assays of intact glomerular filtration barriers (GFB), 3D-Electronic Microscopic reconstruction united with expertise in VEGF, a constitutively expressed podocyte growth factor which has multiple isoforms with contrasting properties. The two permeability parameters: hydraulic conductivity (convective permeability to water) and reflection co-efficient (macromolecular selectivity) will be investigated in glomeruli from our recently described unique transgenic animal line that over-expresses the anti-permeability and glomerular cyto-protective VEGF isoform VEGF165b when crossed with i) animals known to over-express pro-permeability VEGF isoforms; ii) transgenic animals in which chronic podocyte injury occurs spontaneously or can be induced in a dose dependant way. In addition, to relate structure to function, the glomerular filtration barrier region (including sub-podocyte and interpodocyte spaces recently identified in this laboratory) from the same glomeruli from which the physiological parameters have been defined, will be reconstructed in three dimensions at the ultrastructural level. The ability of agents which control VEGF isoform balance at the splicing level to influence functional permeability phenotype will also be investigated in cannulated isolated glomeruli, and human archive tissue will be examined to confirm that deranged VEGF isoform balance is a feature of human glomerular disease. The combination of these highly specialised resources and skills is unrivalled worldwide and will provide a unique opportunity to elucidate how VEGF-isoform balance influences glomerular permeability. This project may allow the development of novel strategies to maintain or re-establish normal glomerular permeability. Our findings may therefore have profound implications, not only for renal patients and those at risk of cardiovascular disease but also for wider clinical fields e.g cancer (as deranged VEGF splicing is also a feature of malignancy).


Research activity codes: 2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: RENAL AND UROGENITAL
Title: The Genetic Basis of Urinary Incontinence in Women
Funder: Medical Research Council

Abstract: Aims: To identify genetic polymorphisms associated with stress and urge incontinence in women, and understand how these influence bladder gene expression, and in vivo bladder function. Objectives: 1. To conduct the first genome wide association study (GWAS) of stress and urge urinary incontinence in women, identifying loci associated with incontinence. 2. To fine map significant loci from the GWAS, identifying the susceptibility genes, and confirming the clinical relevance and in vivo functional effects of relevant SNPs and/or their associated genes. 3. To identify bladder specific expression quantitative trait loci (eQTL), relating significant SNPs to alterations in the bladder transcriptome using an integrative genomics approach. Design: Three main studies will correspond to these objectives: 1. GWAS of stress and urge incontinence using a two stage discovery and replication design, in the UK Twin Cohort, and Bristol ALSPAC Cohort. 2. Fine mapping study using patients with stress and urge incontinence as cases and controls. Women referred for clinical bladder function tests will be genotyped from saliva samples. 3. eQTL mapping using bladder biopsies taken at the time of cystoscopy in women with incontinence, who have been previously genotyped for significant SNPs. Methodology: 1. The GWAS discovery set (UK Twin Cohort, n?8,000) will use existing phenotypes and genotypes (Illumina 660). The replication set (ALSPAC cohort, n?15,000), will use a new mailing of a validated self-completed questionnaire for incontinence phenotyping, as well as existing genotypes (Illumina 610). In both cohorts age, BMI, and parity will be considered as potential mediators. 2. For the fine mapping, loci with genome wide significance will be genotyped in detail (n?400) using a Sequenom MASSarray analyser, and associations measured with clinical diagnosis and quantitative urodynamic parameters, using chi square and multivariate mixed regression. 3. 5-10mg bladder biopsies will be collected and stored in RNAlater (n=34). Gene expression will be measured using Affymetrix Human Genome HgU133 Plus 2.0 microarrays, with validation using qPCR. Scientific and medical opportunities: Urinary incontinence is highly heritable, but no contributing genetic markers have been identified. Genome wide studies have revolutionised the search for the genetic causes of complex diseases. We expect that discovered SNPs will serve as markers of clinical risk for stress and urge incontinence, and through relation to gene expression, and bladder function, will help explain the underlying pathogenesis. The insights provided will potentially offer new drug targets, biomarkers, and preventative strategies.


Research activity codes: 2.1 BIOLOGICAL AND ENDOGENOUS FACTORS
Health categories: RENAL AND UROGENITAL
Title: Reducing pro-inflammatory responses to infection to improve peritoneal dialysis patient outcomes
Funder: Division for Social Care and Health Research (Wales)

Abstract: Renal replacement therapy, as a consequence of end-stage renal failure, is a chronic incurable condition. It presents a significant social and financial burden on healthcare in Wales, affecting a considerable and increasing number of patients who require daily therapeutic intervention. Those patients are dependant on dialysis. Two major forms of dialysis are available, Haemodialysis (HD) and peritoneal dialysis (PD). HD is mainly carried out at hospital-based dialysis units. PD is carried out by patients at home. PD remains an essential daily life-saving treatment modality that offers significant treatment, lifestyle, and quality of life advantages over HD, and it is more cost-effective. Recurrent peritoneal infection, with its concomitant inflammation, remains the major cause of PD therapy cessation. Recurrent infection/inflammation leads to peritoneal membrane damage, that results in treatment failure and transfer to HD. New evidence suggests the possibility that the concerted pro-inflammatory activity of specific sensors of microbes in our immune system might result in excessive inflammation, leading to poor PD outcomes. This proposal aims to evaluate this possibility and test potential specific therapeutic strategies. This work may inform the design of novel, more efficient, therapies to improve clinical outcomes in PD and also other acute and chronic inflammatory conditions.


Research activity codes: 7.3 MANAGEMENT AND DECISION MAKING
Health categories: RENAL AND UROGENITAL
Title: Diagnostic accuracy study of a MALDI-MS biomarker of acute kidney injuryafter femoral fracture surgery
Funder: Chief Scientist Office, Scotland

Abstract: One fifth of the 70,000 UK patients that break their hip annually suffer a loss of kidney function (AKI). AKI can lead to a longer hospital stay, dialysis, and even death. Routine tests detect kidney injury at a late stage when treatment is ineffective. We plan to perform a multicentre clinical evaluation of a new test that identifies AKI early with the aim of reducing the severity and harm of AKI after hip fracture. We are applying for CSO funding for pilot work and a patient involvement event.


Research activity codes: 4.2 EVALUATION OF MARKERS AND TECHNOLOGIES
Health categories: RENAL AND UROGENITAL
Title: Human validation of RIP1 Kinase inhibitor for acture kidney injury (AKI)
Funder: Chief Scientist Office, Scotland

Abstract: Acute kidney injury (AKI) has many causes including infection, dehydration and surgery. It affects 1 in 10 hospital patients in Scotland. It can result in a prolonged hospital stay, increased chance of dying in hospital and long term kidney failure and dialysis. There are no medications to treat AKI available curently. We have proven that a new medication prevents and treats AKI in mice. We want to collect human samples for experiments to provide evidence that this drug will work in humans. Human experiments are required in preparation for a clinical trial. We have developed a method of testing drugs for AKI using human tissue in the lab. This has not been done before. This has a wider benefit to the world-wide research community in providing a human model for the testing of drugs for kidney disease prior to a clinical trial.


Research activity codes: 5.1 PHARMACEUTICALS
Health categories: RENAL AND UROGENITAL