Strategies for targeting the tumour microenvironment using high fidelity 3D culture systems

The heterogenous tumour microenvironment (TME) has been well established in the progression of cancer and patient outcome. In particular, the success of anticancer therapy is influenced by various constituents and intercommunicating networks within the TME. Drug development research often focuses on targeting cancer cells alone without considering all the cellular and non-cellular components of the TME, which plays a crucial role in chemoresistance and disease relapse. Thus, there is a necessity to understand the individual features of the TME in order to deliver a more personalised therapy that improves the rate of successful outcomes while reducing the recurrence of the disease. However, the mechanisms of acquired drug resistance and pharmacokinetics within the tumour still remains elusive. Currently, there is a lack of high-throughput methods suitable to study the functional effects of anticancer drugs on tumour samples where the interplay between cancer cells and the TME remains intact.This thesis embodies the work in the development of the ALTEN (Alginate-based Tissue Engineering) platform, a biomimetic hydrogel system for rapid functional testing of anticancer drugs in explanted tumours. The alginate hydrogels provides a 3D scaffold that resembles the native extracellular matrix (ECM) that preserve the original characteristics of the tumour to permit detailed assessment of the TME and the complex molecular interactions between cell species. Here, we use high-resolution single-cell RNA-seq technologies to analyse the molecular effects of anticancer treatments within the tumour to study the mechanisms of acquired drug resistance.The combination of ALTEN and scRNA-seq technology enable high-throughput and high-resolution screening of tumour explants exposed to cytotoxic agents and immunomodulators. The impact of drug therapies within an intact TME could be evaluated to 1) determine treatment efficacy and 2) characterise drug resistant cancer populations.In summar