Grant: Master's Award: Canada Graduate Research Scholarships - Master's program

Breast cancer (BC) remains a highly prevalent and heterogeneous malignancy, with complex pathology that continues to challenge effective treatment and prevention. Among its subtypes, triple-negative breast cancer (TNBC) is particularly aggressive and difficult to treat. While CRISPR-Cas9 gene editing therapy shows promise, its most efficient form is Cas 9 ribonucleoprotein (RNP), which presents a major hurdle for delivery due to its large size and charge. Focused ultrasound (FUS) in combination with intravascular contrast agent microbubbles (MBs) offers a non-invasive, image-guided method to temporarily modify cellular permeability, allowing large biomolecules to pass through the cell membrane. Despite this emerging technology, its application in BC treatment still remains in the early stages, as preclinical studies are still limited. Therefore, developing a more sophisticated in vitro strategy, such as utilizing a 3D culture, is essential for accurately predicting the physiological impact of FUS-mediated delivery of CRISPR-Cas9 RNPs. We will begin by establishing a biologically relevant co-spheroid model to mimic a solid TNBC tumor. Acoustic parameters will then be optimized for the delivery of CRISPR-Cas 9 RNPs to static co-spheroids. Next, we will focus on replicating these efficiencies under different flow rates and extracellular matrix (ECM) compositions to better simulate vascularized tumor environments. Finally, we will engineer a microfluidic platform capable of FUS-mediated delivery of CRISPR-Cas9 RNPs through functional co-spheroid vasculature, demonstrating a highly physiologically relevant model for evaluating targeted gene delivery within a vascularized tumor microenvironment.