HAVOC

Background/rationale: Despite advances in breast cancer therapies, certain tumors remain lethal due to their ability to evade immune system attacks. This immune evasion is strongly influenced by the tumor microenvironment (TME), which includes tumor endothelial cells (TECs), immune cells, and cancer-associated fibroblasts (CAFs), along with extracellular matrix (ECM). TECs particularly exhibit abnormal phenotypes that promote tumor progression and metastasis by increasing angiogenesis, remodeling 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 tumor-on-chip platform that accurately models the TME, particularly TEC interactions and immune cell dynamics, will allow pre-testing 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 tumor organ-on-chip (TrOoC) platform.

Secondary Aims: To optimize a scalable microfluidic platform for modeling the tumor-specific TME with patient-specific material, allowing for high-throughput and -resolution interrogation of tumor-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 tubeless microfluidic platform to host patient-derived tumor 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 platform will support long-term culture and will be characterized via immunofluorescence, multicolor flow cytometry, cytokine assays, and single-cell RNA sequencing. Personalized therapeutic testing will be conducted using bi-specific antibodies and TME-modulating therapeutics, alone or in combination with chemotherapeutics.

Expected results and potential impact: We anticipate that our TrOoC platform will faithfully replicate central aspects of the TME, enabling detailed studies of tumor-immune interactions and TEC behavior. The platform is expected to provide a robust tool for preclinical testing of anti-cancer strategies with a focus on TME perturbation. 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 tumor types, further increasing its impact in guiding personalized therapeutic recommendations.