Background/rationale: Despite advances in breast cancer therapies, certain tumours remain lethal due to their ability to evade immune system attacks. This immune evasion is strongly influenced by the tumour microenvironment (TME), which includes tumour endothelial cells (TECs), immune cells, and cancer-associated fibroblasts (CAFs), along with extracellular matrix (ECM). TECs particularly exhibit abnormal phenotypes that promote tumour progression and metastasis by increasing angiogenesis, remodelling the ECM, and modulating immune responses. This altered TEC environment creates a barrier to effective immune cell infiltration and presents a significant challenge for current therapeutic strategies.
Hypothesis: We hypothesize that a microfluidic tumour-on-chip platform that accurately models the TME, particularly TEC interactions and immune cell dynamics, will allow pretesting and tailoring of immunotherapeutic interventions.
Primary aim: To establish a human-relevant in vitro model of breast cancer, incorporating the TME and TECs, on a robust and scalable microfluidic tumour organ-on-chip (TrOoC) platform.
Secondary aims: To optimize a scalable microfluidic platform for modelling the tumour-specific TME with patient-specific material, allowing for high-throughput and -resolution interrogation of tumour-TME-immune cell interactions. We will characterize the heterogeneous TME interactome using complementary state-of-the-art technologies.
Moreover, we will conduct personalized testing of standard therapeutic regimens and test the effects of therapeutic bispecifics in combination with existing therapies on-chip.
Methods: This project involves adapting a scalable tubeless microfluidic platform to host patient-derived tumour organoids, TECs, CAFs, and peripheral blood mononuclear cells. Initial optimization will use commercial and in-house derived cell lines, followed by integration of patient-derived cells. The TrOoC platform will support long-term culture and will be characterized via immunofluorescence, multicolour flow cytometry, cytokine assays, and single-cell RNA sequencing. Personalized therapeutic testing will be conducted using bispecific antibodies, alone or in combination with existing TME-modulating therapeutics.
Expected results and potential impact: We anticipate that our TrOoC platform will faithfully replicate central aspects of the breast cancer TME, enabling detailed studies of tumour-immune interactions and TEC behaviour. The platform is expected to provide a robust tool for preclinical testing of anti-cancer strategies with a focus on perturbation of the TME. By enabling personalized testing, the proposed technology has the potential to significantly improve therapeutic strategies and patient outcomes in breast cancers, ultimately contributing to more effective and tailored cancer treatments. In the long run, the platform may also be adapted to other tumour types, further increasing its impact in guiding personalized therapeutic recommendations.