BJA/RCoA Project Grants

University of Aberdeen

Title
Investigation of the potential of negative regulation of nuclear factor kappa B using a novel antibody under conditions of sepsis

Amount
£30,179

Scientific Abstract
Sepsis is associated with dysregulated inflammatory responses, oxidative stress and mitochondrial dysfunction, with activation of nuclear factor kappa B (NFĸB) having a major role through regulation of gene expression of key mediators. A20 is a negative regulator of NFĸB, induced by several inflammatory mediators implicated in sepsis. A20 can interact with A20-binding inhibitors of NFĸB activation (ABINs) to inhibit tumour necrosis factor (TNF)-induced activation of NFĸB and is also referred to as TNF-alpha-induced-protein-3 interacting protein-1 or TNIP1. The human leukocyte antigen class I-specific receptor (LILRB1) is a potential therapeutic target which can raise TNIP1 expression, limiting NFĸB activation and its subsequent effects. We have shown that ligation of LILRB1 in dendritic cells increases TNIP1 expression and inhibits cytokine production. We propose to determine the effect of LILRB1 on TNIP1 expression, NFĸB activation, cytokine responses and mitochondrial function, in different immune cell types under conditions of sepsis. We will use both cell lines and also primary cells from patients with sepsis. This study will determine the potential of targeting LILRB1 to disrupt inflammatory pathways during sepsis and the results may also be relevant more broadly, for example in chronic inflammatory conditions such as rheumatoid arthritis, where TNIP1 has been implicated.

Royal Marsden NHS Foundation Trust, London

Title
Tracking the molecular and metabolic fingerprints of inhalational versus intravenous anaesthesia in breast cancer

Amount
£69,787

Scientific Abstract
Surgery remains the best option to optimize long term survival in many patients with cancer. Surgery can lead to tumour cell dissemination into the circulation. Laboratory data has suggested differential effects of anaesthetic agents on cancer cell growth. We performed a retrospective analysis of over 7000 patients receiving inhalational anaesthesia or propofol for cancer surgery. This demonstrated a hazard ratio for death of 1.46 (1.29-1.66) after propensity scoring and multivariable analysis. Our preliminary data has found increased cancer cell survival with volatile anaesthesia. This application contains a detailed mechanistic study, incorporating in vitro, in vivo, and clinical trial data. In vitro work will focus on cell apoptosis, metabolomics and cell migration looking at key markers of PI3K signalling and epithelial-mesenchymal transition. In vivo work will utilise in vivo imaging and ex vivo analysis to quantitatively analyse metastasis. Immunohistochemical analysis will be performed based on targets from the in vitro work. An open label randomised controlled clinical trial will assess transcriptomic changes in breast tumours and changes in innate immunity in response to volatile versus propofol anaesthesia. We hope that understanding the mechanisms of any effect on cancer cells will help inform the design of a large, prospective randomised controlled trial.

Imperial College, London

Title
Mapping inflammation-induced mitogen- and stress-activated protein kinase 1/2-dependent molecular changes in primary sensory neurons

Amount
£90,123

Scientific Abstract
Peripheral tissue inflammation is a major cause of prolonged pain that available therapies can only partially mitigate. Therefore, mechanisms-based analgesics must be developed, which necessitates the clarification of the molecular mechanisms mediating prolonged pain. We have recently reported that mice lacking the mitogen-and stress-activated kinases 1 and 2 (MSK1/2) do not develop inflammatory heat hypersensitivity. While our pilot data show that inflammation induces MSK1/2-dependent changes in primary sensory neurons (PSN) whose activity constitutes the "driving force" for nociceptive signalling and the development of prolonged pain in the central nervous system, the full molecular mechanisms of MSK1/2-mediated effects on cell signalling and gene transcription remain to be elucidated. Here, we will investigate the MSK1/2-dependent and independent signalling and transcriptional changes in PSN following capsaicin-induced inflammation in wild type and MSK1/2-deficient mice. Combined immunostaining will be used to find cell specific expression of MSK1/2's effectors, whereas RNA sequencing, chromatin immunoprecipitation followed by DNA sequencing, and bioinformatics will be used to identify genes whose expression is altered in MSK1/2-dependent and independent manner in dorsal root ganglion neurons. Findings of this study will improve our understanding of nociceptive mechanisms in PSN and will serve as pilot data for further grant applications.

Craigavon Area Hospital, Belfast

Title
Determining the mechanisms by which active deresuscitation in stable critically ill patients may modulate clinical outcomes

Amount
£49,731

Scientific Abstract
The optimal approach to fluid balance in critically ill patients is uncertain. In a recent systematic review, we found low quality evidence in favour of a conservative fluid or deresuscitative approach (active removal of accumulated fluid using diuretics and/or renal replacement therapy) compared with a liberal strategy or usual care. We will undertake a pilot randomised trial (RADAR-2) comparing conservative fluid and deresuscitation with usual care in patients who are mechanically ventilated and require treatment in intensive care for at least 48 hours. The primary outcome will be a difference in fluid balance between the two arms on day 3, with several secondary feasibility and safety outcomes, including long-term cognitive function.

We are seeking funding to investigate the mechanisms by which fluid strategies may modulate clinical outcomes. We aim to test the hypotheses that a conservative/ deresuscitative fluid strategy:
1. Does not adversely affect regional cerebral oxygen saturation and post-ICU cognitive dysfunction
2. Improves cardiac function and reduces endothelial injury

These findings will inform the design of future large randomised trials of alternative fluid strategies and will provide insights into the physiological effects of fluid overload, and to potential risks and benefits of an intervention to minimise fluid overload.

Imperial College, London

Title
Investigation of the cyclophilin A:CD147 axis as a novel mediator of ventilator-induced lung injury

Amount
£69,847

Scientific Abstract
Ventilator-induced lung injury (VILI) plays an important role in determining morbidity and mortality of patients in the intensive care unit. Despite much pre-clinical research, no pharmacological intervention has yet proven effective to limit VILI in patients. Within the last 10 years, secreted cyclophilin A (CypA) and its receptor CD147 have emerged as a novel signalling axis in various diseases. We recently demonstrated that high fat-fed mice, which were protected from VILI, showed attenuated expression of CD147 within their lungs. The current study is therefore designed to explore whether the CypA:CD147 axis plays a previously unexplored role during the pathogenesis of VILI.

We will initially investigate the basic biology of CypA:CD147 in lung cells using in vitro models of lipopolysaccharide-induced inflammation and cell stretch, as there is currently no mechanistic understanding of the axis within the lungs. We will then explore the nature of CypA:CD147 signalling using in vivo models, followed by experiments to investigate the consequences of CypA:CD147 blockade using clinically-relevant mouse models of VILI.

These studies will substantially enhance our understanding of the CypA:CD147 axis during lung inflammation and injury, and may potentially point towards the axis as a novel therapeutic target for ventilated patients.