Novel imaging strategies to investigate 3D tumour invasion at the molecular level

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Cancer is the one of the major causes of deaths worldwide and despite our increased understanding and the development of more improved treatments, many advanced cancers are still incurable. The majority of these deaths are due to metastasis whereby through cascading events, cancer cells escape the primary tumour, acquiring cellular characteristics which enable them to colonise other parts of the body. Several underlying mechanisms of this process are not understood and require the development of new biological models which can mimic cellular interactions within these complex tumour ecosystems. Until now most of our understanding of the basic molecular principles of cancer have come from experimentation with single layer 2D cell culture models. Whilst inexpensive and easy to image, they are not representative of the way cells actually interact in an in vivo environment. 3D cell cultures (i.e. spheroids), have gained great significance in recent years as they can provide a much more realistic environment to simulate biological interactions. Unfortunately as the relative size, heterogeneity and hence complexity of these 3D cell cultures increases, it imposes severe limitations when imaging with any optically based technique. Local variations in refractive index induce optical aberrations and scattering, and coupled with absorption, lead to severe a degradation in optical resolution, as well as a loss in signal and contrast. The key objective of this fellowship is to develop novel optical imaging strategies which will enable for the first time, the complete interrogation of tumour derived spheroid models, to understand how cancer cells to dissociate from the primary tumour, evade immune surveillance and invade surrounding tissues. The imaging platform will be based on light sheet fluorescence microscopy (LSFM) to enable high-speed volumetric imaging capability. Building on previously conducted research at King's College London, the system will incorporate fluorescence lifetime imaging (FLIM) to allow functional information to be extracted, which will be instrumental in gaining an understanding of the molecular interactions taking place. Several aspects of the design will be considering including the incorporation of adaptive optical elements (i.e. spatial light modulator) to counteract aberrative effects in both the illumination and detection paths. Using this technology and working in close collaboration with a number of life scientists, tumour spheroid cultures will be constructed to closely imitate the tumour microenvironment (TME). Several aspects of this TME will be examined including (i) cell motility and migration of cancer cells and (ii) their effects on the structural modelling of the extra cellular matrix (iii) the role of immune cells in tumour progress.Analysis of this multivariate and multidimensional data represents a huge challenge and a number of conventional machine learning and deep learning techniques will be explored. The development of instrumentation to image, extract, and analyse functional information of large 3D cell culture models non-invasively, at high-speed and at high resolution is vital and will further improve our understanding of cellular complexities of interactions between cells and the Extracellular Matrix (ECM). This will serve to assist in the construction of more complex spheroid structures to mimic in vivo conditions and has enormous potential to aid in the development on specific anti-cancer therapies, revolutionising patient treatments and outcomes.