University of Southampton

2013 – 2017

Institute of Developmental Science at Southampton General Hospital

The research projects I was associated with include:

1. Improving extracellular vesicle resolution and phenotyping using high-throughput techniques.

This project is based around the development of extracellular vesicle analysis equipment, and has led to collaboration with industry to develop high-resolution, high-throughput analysis platforms. These platforms aim to have a dynamic range capable of detecting single extracellular vesicles, by means of scatter and fluorescence, in the size range of 30-1000nm. The development of these high resolution platforms will make research into extracellular vesicles easier and more accurate, and aid their applications in diagnostic and therapeutic medicine along with our understanding of their role in physiology.

2. Analysing the effect of an EPA double-blind placebo trial on non-alcoholic fatty liver disease (NAFLD) patients using microvesicles as biomarkers

NAFLD is the most common chronic liver disease, frequently associated with diabetes. Both of these insulin resistant states have increased cardiovascular risk factors associated, and a prevalent cause of mortality in these diseases. Both are associated with multi-organ inflammation, including the vasculature, immune cells and platelets, potentially altering microvesicle production and consequently the impact of microvesicles on cellular behaviour.This is reflected in the microvesicle studies that have been conducted to date, using traditional markers, which show increases in microvesicle numbers derived from the endothelial cells, white blood cells and platelets. This project will provide evidence for the efficacy of EPA as a treatment for NAFLD as well as providing data on the variation in microvesicle subsets for the disease, which are currently lacking.

3. Can circulating leukocyte microvesicles be used as an early predictor of subclinical infection after acute ischemic stroke?

Infection occurs within the first week in up to 50% of stroke patients, with increased mortality and reduced quality of life in survivors. Leukocyte microvesicles (MVs) are submicron membrane vesicles, produced when white blood cells are activated. During this PhD, I will develop novel flow cytometry based methods for leukocyte MVs. These new methods will examine leukocyte MV changes in stroke patients to investigate if MVs can predict the development of infection, before clinical symptoms arise. Transferring these new methods into a novel biomedical device suitable for finger-prick blood samples will then be investigated. This multi-disciplinary project draws upon the expertise of established teams of researchers from the Faculties of Medicine, Physical Sciences and Engineering, as well as the NHS. The techniques used will include flow cytometry, cell culture, working with clinical samples, ELISAs, and biophotonics.

4. What is the role of extracellular vesicles in breast milk in the early life programming of immune responses and the development of allergy?

Breast milk contains factors that influence the development of the baby’s immune system. These factors may include microvesicles, which are found in the breast milk and have been show to contain high levels of immune related microRNAs. These microvesicles are very stable in the harsh conditions found during digestion, and may give a clue to a mechanism whereby immune-regulating material is passed from the mother to the baby at a stage where the babies immune system is immature and susceptible to early life programming. Novel high-resolution flow cytometry, and genetic profiling techniques will be employed to study leukocyte extracellular vesicles. We will use these new methods to examine leukocyte extracellular vesicles in breast milk from mothers whose infants go on to develop allergy, compared to those infants who do not.