Integrated multi-dimensional molecular organ imaging

Lead Research Organisation: University of Edinburgh
Department Name: MRC Centre for Inflammation Research

Abstract

The development of innovative optical imaging technologies has led to a revolution in light microscopy heralding the development of a generation of microscopes that promise powerful new applications for biomedical research. We believe our plans for refinement of commercially available instrumentation, development of novel probes and comprehensive testing in complex biological applications will ensure our advanced light microscopy 'toolkit' can be exploited to deliver innovative research and improved health outcomes.
To maximise our contribution to the innovation and application of advanced imaging we focus on intra-vital imaging as this platform allows detailed investigation of biological events at sub-cellular resolution in living specimens at the greatest possible penetration depth. We will deploy a state-of-the-art intra-vital light microscope to acquire images of specifically labelled or label-free molecules up to one millimetre into tissue(s) of live animals typically mouse or zebrafish. Our innovative approach includes complementing these images with 3D mapping in order to provide a much more complete view of organ function in health and disease.
Effective, informative, methods for labelling cells prior to intra-vital imaging remains a significant challenge in the wider application of these methods to non-transgenic models. In order to increase the reach of our intra-vital imaging methods we will employ and evaluate novel fluorescent 'Smartprobes'. These Smartprobes are already being used in Edinburgh to image key biological and pathological events in human lungs through cutting-edge fibre-optic microscopy, thus providing a direct translational pathway from planned studies in zebrafish and mice to human diseased organs in life (the technology is readily applicable to the digestive tract and the reproductive system). Our group has already established a strong track record in the use of a wide range of model animals that express fluorescently labelled molecules of interest in key target cells/organs. This expertise will guarantee we develop robust methodology focussed on scientifically challenging questions, such as: What is the involvement of macrophages in the development of breast cancer? How do neurons and glial cells interact in the development, function and repair of the nervous system? How do sex steroid hormones alter cell behaviour in the uterus and how can this be modulated? What is the distribution of central neuronal proteins in the brains of normal compared to model mice suffering from brain disorders?
Excitement surrounding the application of Intra-vital imaging to biomedical research is based on its capacity to track cells and their contents within live tissues offering a novel platform for the study of dynamic changes in protein distribution, molecular interactions and tissue composition. Currently, such observations are only possible at single locations within a tissue - our goal is to expand data-acquisition to whole organs, e.g. to interrogate the distribution of synaptic proteins in a mouse brain. To achieve this, we will integrate our intra-vital techniques with systematic 3D image acquisition on a fast confocal microscope, capable of scanning whole sectioned organs. The resulting series of 3D-images will be archived to ensure data preservation and will be made available to the wider research community via searchable web-based organ libraries. These molecular 'organ maps' detailing the location of specific labelled proteins will allow us to refine subsequent 'focussed' functional intra-vital imaging and provide high-content 3D organ maps with functional measurements from live specimens.
These innovative studies will provide data for our translational research pipeline and inform development of improved diagnostics and therapies for health problems including cancer, multiple sclerosis, infertility, cardiovascular and neurodegenerative diseases.

Technical Summary

The next wave of biomedical science requires 'next generation' LM imaging and new modes of translating it into clinical practice. Our new technology platforms will draw on existing collaborations with commercial partners and physical scientists (chemists, engineers, optical physicists and IT specialists) from UoE and elsewhere. A 'true' multi-functional two-laser intra-vital imaging platform will be developed in collaboration with La Vision BioTech. This will be enhanced to provide a platform that allows simultaneous photo-ablation, -uncaging, -activation, -conversion, -switching, fluorescence lifetime (FLIM) and non-linear optical measurements of relevant cells, proteins and small molecules. We will develop and integrate adaptive optics for tissue aberration correction and a CARS element to enable label-free high resolution imaging of lipid- and collagen-containing structures. We will also incorporate the 'spinning disk' technology and IT systems that have allowed 'beyond-state-of-the art' whole brain mapping and extend this to other organs. This unique facility will enable our campus, with 3 MRC Centres, not only to build 'information-dense' 3D gene expression maps of whole organs, but also provide complex biological data using multiple fluorescent probes and 'label-free' CARS simultaneously. Catalysed by the adjacency of the Edinburgh Royal Infirmary, our interdisciplinary Centres share the challenge of translating their work into the diagnosis and treatment of human disease. This will be facilitated by collaboration with Haslett/Bradley who are synthesising and validating bespoke fluorescent 'Smartprobes' against key inflammation/scarring targets and using a unique pCLE platform to translate into human lung disease in vivo. Molecular Imaging is a Core theme in Edinburgh which has identified the optimal site for the proposed facility, the costs of its fitting out and a major 'cash' contribution towards the equipment.

Planned Impact

Our innovative research programme focuses on the development of integrated multi-dimensional molecular organ imaging promising a step change in our understanding of the molecular organization of whole organs in health and disease.

The planned programme of research and innovation focuses on development of technologies improving the capacity biomedical research 'Changing Lives' by delivering benefits to all sections of society.

The knowledge gained from addressing the technological challenge of mapping proteins in both time and space (4D) will be of immediate interest and benefit to industry and scientists researching the initiation and progression of disease. Details of technical innovations related to instrumentation will be of immediate interest to companies who manufacture and develop optical microscope systems, and components, and the associated software required for high-end image storage and analysis. To ensure information generated during the course of the planned programme of work achieves maximum reach within the biomedical community we will ensure all data is preserved and that image datasets and their analyses are made available on publicly accessible web-sites.

The new knowledge gained from mapping of whole organs will benefit clinicians in the NHS and commercial organisations developing new strategies for treatment of patients suffering from debilitating conditions such as mental ill-health, multiple sclerosis, Alzheimer's, heart disease, infertility and cancer that impact on millions of people in the UK and world-wide. For example, the planned studies on the brain including mapping dynamic associations between proteins in brain synapses and signals guiding microglia will inform strategies for adjusting therapies to improve responses to drug treatments of mental disorders and treatment of brain injury. In addition, a multifaceted approach to the study of macrophages in the tumour microenvironment including use of novel smart probes will inform therapeutic approaches to inhibit tumour progression and malignancy.

Information gained during the course of this programme will be of interest to patient groups and the wider public. We plan to disseminate our findings beyond the academic and commercial sectors by using web-based and other media to inform the general public, patients and special interest groups, charities, local and national politicians about our progress.

Our plan of work offers opportunities for training and capacity building in biomedical research and technologies related to image acquisition and analysis that will have a long-term benefit on health and wealth creation.
 
Description EMBO Long-Term Fellowship Award to Susan van Erp
Amount £120,000 (GBP)
Organisation European Molecular Biology Organisation (EMBO) 
Sector Learned Society
Country European Union (EU)
Start 11/2016 
End 10/2018
 
Description PhD studentship scheme
Amount £85,000 (GBP)
Organisation Alzheimer’s Society 
Sector Charity/Non Profit
Country United Kingdom of Great Britain & Northern Ireland (UK)
Start 03/2016 
End 02/2019
 
Description Wellcome Trust Investigator Award to Prof Peter Brophy, CNR
Amount £1,349,913 (GBP)
Funding ID 107008/Z/15/Z 
Organisation The Wellcome Trust Ltd 
Department Wellcome Trust Senior Investigator Award
Sector Private
Country United Kingdom of Great Britain & Northern Ireland (UK)
Start 03/2016 
End 02/2018
 
Title Intravital optical measurement of physiological parameters in mice (Horsburgh group) 
Description Light microscopy-based, intravital measurement of red blood cell velocity in live mice; molecular imaging of surface anitgens and leukocyte rolling and adhesion; amyloid accumulation in vivo in longitudinal studies 
Type Of Material Model of mechanisms or symptoms - mammalian in vivo 
Provided To Others? No  
Impact The further development of these techniques allowed the measurement of very important parameters in the brains of live mice. This includes the three-dimensional characterisation of amyloid accumulations in vivo (mice).