Miniaturised Metabolomics Platform for Microvascular Research
METAFORA is an individual fellowship awarded to the experienced researcher Dr. Yuliya Shakalisava by the Marie Skłodowska-Curie Actions of the European Commission. This fellowship supports a career re-start in research after an extended career break through the technical project and an individual training programme. The fellowship takes place at Analytical BioSciences and Metabolomics group, Leiden Academic Center for Drug Research, Leiden University, The Netherlands.
Microvascular disease is a chronic diseases which lacks efficient treatment because the disease mechanism is still unclear. Microvascular disease is the result of continued exposure of the small blood vessels in human body to high blood pressure, obesity and diabetes. It leads to the progressive loss of tissue capillaries, tissue-ischemia and fibrosis. As a consequence, the damaged vessels is a main cause of morbidity of the elderly, leading to end-organ diseases including heart failure, kidney failure, dementia, and beta cell failure in pancreas. Worldwide, approximately 80% of all diabetic patients develop retinopathy, 20% develop neuropathy, and 40% develop nephropathy. A novel approach is needed to study the pathology of microvascular disease.
In the last 10 years extracellular vesicles emerged as a new target in disease biomarkers and the keys to disease mechanisms. These nanosized vesicles are produced by all cells and represent an information rich matrix containing proteins, mRNA and metabolites of the parental cell. The exosomes can be found in all body fluids (blood, urine, tears) and that is why they are possible to access without invasive procedures. There is increasing evidence that detecting blood extracellular vesicles of epithelial and platelet origin may present very useful non-invasive signature for the onset of microvascular disease. The application of metabolomics methodologies and extracellular vesicles to the study of microvascular disease can increase our understanding of the pathophysiological processes involved and this should help to identify potential biomarkers to diagnose disease and develop new therapeutic strategies. Isolating and separating extracellular vesicles from blood, however, is very challenging. This project proposed a new way to resolve this problem.
In this project two novel approaches of separating extracellular vesicles from bio fluids were investigated. An electrodriven separation in the conventional capillary electrophoresis format was achieved for the analysis of plasma nanoparticles. A microfluidic chip format was also developed and applied to the exosomes samples. Both methods allow to collect individual fractions of nanoparticles and perform follow up metabolomic analysis.
The approach developed in this project was completely novel for this area of research and is based on the physical properties of nanoparticles such as electrical charge and mass. This method will help to investigate the microvascular disease in a new way and, hopefully, will lead to the discovery of new disease biomarkers. The work will continue beyond this project.