Microfluidic Microwell Platforms for Drug Response Studies in NSCLC Patient-derived Organoids

Lung cancer is the most common type of cancer worldwide, while non-small cell lung carcinoma (NSCLC) accounts for 85% of all lung cancer diagnoses and has a less than 26% 5-year survival rate in the US. Targeted therapies have shown promise in NSCLC patients with specific mutations, but drug resistance and acquired mutations reduce their effectiveness. Therefore, identifying personalized NSCLC treatments is crucial. Patient-derived organoids (PDOs) are valuable preclinical tumor models due to their ability to retain original tumor heterogeneity. However, limited sample availability, slow growth, and size variability in current culture platforms impede PDO's therapeutic application. To address these challenges, we developed a novel microfluidic microwell platform that ensured PDOs' healthy condition and size uniformity, simulated fibroblast-induced drug resistance, and achieved high-throughput screening. In this thesis, we initially harnessed 3D-printed U-shaped agarose microwells (AMWs) to establish a proof-of-concept platform for cultivating and screening cancer cell spheroids and PDOs in an open system. Our results demonstrated the effectiveness of U-shaped AMWs in generating spheroids and PDOs of uniform size, remarkable viability, and clinically relevant drug responses. Fibroblast-induced drug resistance was then introduced to tumor models to simulate tumor microenvironment impacts on drug resistance. Finally, a multi-channel microfluidic device was utilized for high-throughput drug screening on a complex tumor model, which included PDOs with fibroblast-induced resistance and microfluidic chip-based perfusion.