Single and Combination Drug Screening with Aqueous Biphasic Tumor Spheroids

摘要

Spheroids of cancer cells represent a physiologic model of solid tumors for cancer drug screening. Despite this known benefit, difficulties with generating large quantities of uniformly sized spheroids in standard plates, individually addressing spheroids with drug compounds, and quantitatively analyzing responses of cancer cells have hindered the use of spheroids in high-throughput screening applications. Recently, we addressed this challenge by using an aqueous two-phase system technology to generate a spheroid within an aqueous drop immersed in a second, immiscible aqueous phase. Integrating this approach with robotics resulted in convenient formation, maintenance, and drug treatment of spheroids. Here, we demonstrate the feasibility of high-throughput compound screening against colon cancer spheroids using 25 anticancer compounds. Using a strictly standardized mean difference and based on a preliminary testing with each compound, we select effective compounds for further dose-response testing. Finally, we use molecular inhibitors to target upregulated protein kinases and use them for drug combination studies against spheroids. We quantitatively analyze the combination treatment results using statistical metrics to identify synergy between pairs of inhibitors in compromising viability of colon cancer cells. This study demonstrates the utility of our spheroid culture technology for identification of effective drug compounds, dose-response analysis, and combination drug treatments.