Identifying changes in proteome of single cells using imaging mass spectrometry

Principal Investigator: Professor Doug Turnbull

This project aims to develop a new technique enabling the analysis of expression of multiple proteins in single cells in situ.

The Study

The main aim of Newcastle Biomedical Research Centre (BRC) is to improve the lives of patients with long term conditions and ageing syndromes. All the themes of the BRC are planning to develop new treatments. To monitor the effect of treatment, tissue samples are often taken. These samples are then analysed in the laboratory using techniques to identify the cause of disease and why a treatment may not be working. The techniques we currently use are very sensitive and involve looking down a microscope at proteins which are labelled with probes that fluoresce– meaning we can detect the changes in four different proteins all at the same time on a tissue sample. However the current methods are limited because they are detected using light with four proteins being the maximum number. This grant is to use a technique which is ten times better and involves using probes which we detect using their weight – this means rather than four proteins we can detect up to 40 proteins on the same section. This extra information is critical for us developing new treatments and new biomarkers of disease activity.

This work will not only greatly increase our understanding of long term conditions and ageing syndromes but also improve the way we diagnose and treat patients.

Next steps

A major advantage of our proposal to fund the development of IMC across the BRC is that there is already major funding in place from MRC, Wellcome and CRUK for further studies using IMC. It is also planned that IMC will be one of the methods to evaluate tissue from forthcoming clinical trials for example the recently funded MRC DPFS award for treatment of mitochondrial myopathies. In addition, IMC will play a central role in the evaluation of muscle biopsies from a planned life course study of age related changes in muscle.
The Hyperion IMC unit will be housed in the well-established Newcastle Flow Cytometry Core Facility (FCCF) and will seamlessly integrate with the already established and functioning Helios CyTOF system. The Hyperion system will be supported along with the existing extensive expertise in Mass Cytometry (Helios) by Dr Andrew Filby and his staff. The Hyperion technology will be integrated in to a local “Newcastle BRC image “Tissue” Cytometry” user group initially comprising the PI and co-applicants. Access to the system will be charged at an hourly cost recovery rate in line with the existing core facilities policy. This will ensure the sustainability of the research.

Project milestones

• Optimise Hyperion system on muscle sections using antibodies already labelled with lanthanides, and publish first paper using the methodology in the diagnosis of mitochondrial muscle disease.
• Prepare a catalogue of validated antibodies for use across the Newcastle BRC including antibodies to cellular pathways, for detection of individual cell types and for cell membranes. Also optimise/validated the antigen retrival and staining conditions for these antibodies in different tissue settings under different preservation conditions (frozen, FFPE etc).
• Use in the evaluation of clinical trials for long term conditions.
• Develop new clinical diagnostic pathways and new therapeutic approaches.
• Comprehensive analysis of muscle biopsies from individuals through the life course and in patients with sarcopenia.