Project title: New paradigm of GPCR signalling at intracellular sites in metabolic diseases
Supervisors: Professor Davide Calebiro (University of Birmingham), Dr Stephen J. Hill (University of Nottingham) and Professor Gareth Lavery (University of Birmingham)
G protein-coupled receptors (GPCRs) mediate the effects of several hormones and neurotransmitters and are major pharmacological targets. Whereas GPCRs were long believed to signal exclusively at the plasma membrane, our recent findings indicate that these receptors signal also at intracellular sites, such as early endosomes or the Golgi. In this project, advanced live-cell imaging methods, such as FRET/BRET, FCS and single-molecule microscopy, will be combined with state-of-the-art metabolomics to investigate the role of GPCR signalling at intracellular sites in adipocyte metabolism. We expect these experiments to lead to a deeper understanding of the mechanisms involved in the regulation of adipocyte metabolism and to the identification of new molecular targets for the therapy of metabolic diseases. The successful applicant will join a vibrant, dynamic and internationally-recognised team at the Institute of Metabolism and Systems Research and at the Centre of Membrane Proteins and Receptors of the Universities of Birmingham and Nottingham.
Project title: Identifying platelet derived proteins that regulate the thrombo-inflammatory recruitment of monocytes in atherosclerosis.
Supervisors: Professor Ed Rainger (University of Birmingham), Professor Alison Goodall (University of Leicester) and Dr Myriam Chimen (University of Birmingham)
It has long been appreciated that platelets play an important role in the symptomatic stages of cardiovascular disease by contributing to the formation of the thrombus which precipitates heart attack and stroke. However, we now realise that platelets also interact with the inflammatory response to support initiation and development of atherosclerotic plaque long before symptomatic disease is apparent. Why is this important? Because platelets did not evolve as key cellular components of the immune and inflammatory responses, meaning that their contributions (thrombo-inflammation) are not appropriately regulated and can result in pathology. We now believe that platelet activation and shedding of platelet derived extracellular vesicles (PEV) is a major communication route between leukocytes and monocytes in the circulation. In this study we will identify the ‘cargo’ transferred from platelets to monocytes via PEV and using in vitro and in vivo assays, determine how this regulates monocyte recruitment and migration.
Project title: Exploration of the global manipulation of transcriptional networks by oncogenic human papillomavirus
Supervisors: Dr Joanna Parish (University of Birmingham) and Dr Sally Roberts (University of Birmingham)
Human papillomaviruses (HPV) cause cancers of the anogenital and oropharyngeal tracts. The HPV life cycle is dependent on the differentiation of infected keratinocytes. Infection is established in the undifferentiated basal keratinocytes where the chromatinised viral DNA is maintained and viral transcripts encoding viral oncoproteins are expressed. Differentiation of cells within the epithelium coincides with activation of the late virus promoter and capsid protein synthesis. As well as controlling expression of their own genes, viruses create a host environment that supports replication. The mechanisms of virus-mediated transcriptional reprogramming during the HPV life cycle are not understood but we hypothesize that HPV epigenetically reprograms the host to create a cellular milieu supportive of viral persistence and these changes contribute to HPV-driven carcinogenesis. The student will use state-of-the-art models of HPV-infected tissue, and advanced technological methods to dissect genome wide host manipulation during infection and determine those changes that contribute to HPV-driven cancer.
Project title: Understanding how tetraspanins and the ‘molecular scissor’ ADAM10 promote blood cancer
Supervisors: Dr Mike Tomlinson (University of Birmingham), Dr Steve Briddon (University of Nottingham), Dr Farhat Khanim (University of Birmingham) and Professor Nick Holliday (University of Nottingham)
T-cell acute lymphoblastic leukaemia (T-ALL) is an aggressive blood cancer that is in urgent need of more effective therapies. Over 65% of T-ALL is driven by activating mutations in the cell fate regulator Notch1, which allow its activation by the ‘molecular scissor’ ADAM10. We have discovered that ADAM10 substrate specificity is dictated by its association with one of six regulatory tetraspanins - Tspan5, Tspan10, Tspan14, Tspan15, Tspan17 and Tspan33. We propose that ADAM10 is not one scissor, but six different scissors depending on the associated tetraspanin. The aims of this project are to determine (1) which tetraspanin(s) promotes cleavage of mutant Notch1 in T-ALL, (2) whether cleavage occurs at the cell surface or on vesicles following endocytosis, and (3) whether targeting TspanC8s with antibodies can inhibit proliferation of T-ALL cell lines and primary cells from patients. Experimental techniques will include CRISPR/Cas9 genome editing and advanced fluorescent microscopy.
Project title: The human gut microbiome as a reservoir of antibiotic resistance
Supervisors: Professor Willem van Schaik (University of Birmingham) and Dr Sarah Kuehne (University of Birmingham)
The human gut harbours a complex microbial community (‘the gut microbiome’), which contributes to human health. Previous work by Professor Van Schaik has revealed that the gut microbiome comprises a large number of antibiotic resistance genes. Current methodologies to study the gut microbiome do not allow for the identification of the bacteria that carry these resistance genes. The extent by which resistance genes can spread in gut microbiome also remains unclear.
In this project, you will use innovative experimental approaches to determine the bacterial reservoirs of antibiotic resistance genes in the gut microbiome. Strains harbouring resistance genes will be cultured and characterized by genome sequencing. Finally, conjugation assays will be performed to assess the ability of these bacteria to serve as donors of resistance genes to opportunistic pathogens.
This project will importantly deepen our understanding of the mechanisms that contribute to the rapid spread of antibiotic resistance in bacterial ecosystems.
Project title: Does obesity-associated synovitis promote joint pain in osteoarthritis?
Supervisors: Dr Simon Wyn Jones (University of Birmingham) and Professor Victoria Chapman (University of Nottingham)
Generic analgesics for OA patients provide minimal relief and are associated with severe side effects, emphasising the need for a disease-specific targeted approach. In knee OA, distinct patterns of inflamed synovial tissue (synovitis) are associated with pain and synovitis promotes OA pain severity due to the sensitizing action of inflammatory cytokines on joint nociceptors.
We recently reported that the synovial fluid of obese OA patients is more inflammatory compared to normal-weight patients, with the cells of the joint lining (synovial fibroblasts) secreting more pro-inflammatory cytokines. This suggests that the intrinsic phenotype of the obese OA synovial fibroblast is a major contributor to the inflammatory OA joint environment. This project will therefore determine whether obesity-associated synovitis promotes joint pain in OA and determine the functional role of inflammation-associated RNA transcripts in mediating OA pain.
Project title: Understanding of metabolism and functions for non-coding RNA in Prader-Willi syndrome
Supervisors: Dr Pawel Grzechnik (University of Birmingham) and Professor Chris Bunce (University of Birmingham)
The goal of the project is to understand transcriptional processes leading to neuro-developmental disorder known as Prader-Willi syndrome (PWS). PWS it is the most common syndromal cause of life-threatening obesity in humans. PWS patients display broad pathological spectrum including dysmorphic changes, behavioural problems and mild intellectual disabilities. The syndrome results from the loss of expression of various chromatin associated, non-coding RNAs synthesized from the PWS region. Thus, the aim of this project is to investigate how non-coding RNAs transcribed from the PWS locus affect global transcription and chromatin organization. Elucidating of molecular mechanisms of Prader-Willi syndrome will help to better understand roles for non-coding RNA in the development of the human body. The project will employ a wide variety of molecular and cell biology techniques including genome editing, RNA sequencing, chromosome conformation capture techniques and RNA and protein visualization in situ.
Project title: Identifying Novel Therapies to Prevent Atherosclerosis
Supervisors: Professor Roy Bicknell (University of Birmingham) and Professor Ed Rainger (University of Birmingham) and Dr Asif Iqbal (University of Birmingham)
There is intense interest in identifying the genes that predispose to the development of atherosclerosis. We recently carried out an expression analysis of human tissue using microarrays that identified several candidate genes. Generating knockout mice and crossing them to apoE knockout mice followed by placing the double knockout mice on a high fat diet confirmed that at least two of the genes strongly promote atherosclerosis. The aim of this project is to examine whether blocking of these genes using several molecular approaches can prevent development of the atherosclerotic plaques in mice.
That our novel genes are major promoters of atherosclerosis and that blocking their activity can prevent the development of plaques in ApoE knockout mice.
Experimental Methods and Research Plan
Experimental Methods to be learned in the project: Basic molecular biology, DNA cloning, protein expression, gene expression analysis using microarrays, mouse models of disease, genetic alteration of mice using CRISPR, mouse pathology, histochemical tissue analysis
Project Title: Understanding how extracellular environmental factors promote altered intracellular signalling and gene expression networks using transcriptomics in tumour models: development of Cholangitis and Cholangiocarcinoma
Supervisors: Dr Padma-Sheela Jayaraman (University of Birmingham) and Professor Anna Grabowska (University of Nottingham)
Invasive Cholangiocarcinoma (CCA) is an aggressive cancer of the bile duct with very poor prognosis and few treatments. Primary Sclerosing Cholangitis (PSC) is a chronic inflammatory disease of the bile duct where there is no effective treatment other than liver transplantation. New treatments for both are urgently required.
The Proline Rich Homeodomain (PRH/HHEX) protein is a transcription factor with a role in tumour formation in a variety of cancers. PRH is essential for liver and bile duct development. PRH regulates gene expression using multiple mechanisms including recruitment of epigenetic regulators. PRH protein levels are altered in CCA and we wish to investigate the hypothesis that alterations in PRH proteins are regulated by the extracellular environment through activation of, inflammatory or bile acid signalling pathways or through stromal cell interactions. To understand the role of the extracellular environment we will use a combination of cell lines, primary bile duct tissues, in vivo mouse models and primary xenograft derived tissues coupled with transcriptomics approaches.