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

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 recently 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, organoids), 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 original application aimed to enhance light sheet fluorescence microscopy (LSFM) with fluorescence lifetime imaging (FLIM) capabilities. LSFM enables high-speed volumetric imaging essential for observing the dynamic behaviour of cancer cells. Incorporating FLIM allows extraction functional data from these images, providing insights into the molecular interactions within spheroids.
In the fellowship renewal, I plan to take these capabilities further by focusing on more complex tissue models that resemble aggressive cancer types like triple-negative breast cancer. Using advanced light-sheet FLIM microscopy, I aim to achieve a more detailed understanding of cancer cell behaviours and interactions within these models.
A key advancement in our research is the integration of three-photon excitation into our advanced FLIM microscopy platform. This technique will enable deeper penetration into 3D cell cultures, allowing exploration of spheroids and organoids beyond the limitations of current imaging methods. The ability to visualize deeper structures within these models opens new avenues for understanding the mechanisms of cancer cell behaviour.
Moreover, the scope of this research extends to other areas of biology, including developmental biology, neuroscience, and cardiovascular research. The imaging technologies have broad applications and the potential to provide novel insights into various biological processes, enhancing our understanding of disease mechanisms and potential therapeutic approaches.
Overall, this project represents a significant advancement in optical imaging, with the potential to impact not just cancer research, but also a wide range of biological and medical fields. By pushing the limits of our imaging capabilities, Iaim to deepen our understanding of complex biological systems and contribute to the development of new treatments and strategies in healthcare.