Development of Microfluidic 3D Apoptotic Cell Arrays for Anti-cancer Drug Screening and Apoptotic Signaling Profiling
Sihong Wang, PhD and Xuejun Jiang, PhD
The promise of improved cancer therapy has been one of the driving forces for cell death research over the past decade. There is growing evidence that many of the molecular and cellular changes that occur in cancer development diminish the ability of cells to undergo apoptosis and that resistance to apoptosis causes drug resistance. On the other hand, many studies have demonstrated that apoptosis is a frequent outcome of effective anticancer therapy. Therefore, developing and screening novel anticancer drugs that target apoptosis pathways have received increasing attention in the past few years. However, identification of novel compounds and drug targets involved in apoptosis regulation is still a major roadblock to anticancer drug development due to the lack of a high throughput apoptotic screening system which can systematically measure dynamic expression of multiple proteins and genes as well as enzyme activities in real time in intact cells from multiple stimuli. The overall goals of this project are: a) to develop a functional high throughput apoptotic cell array for anti-cancer drug screening and mechanism discovery by combining advanced technologies in BioMEMS with current knowledge in cancer biology and cancer pharmacology and b) to understand cellular and molecular changes in solid tumor and corresponding nonmalignant cells after treatments of clinical and potential anticancer drugs targeting apoptotic signaling pathway systematically with a high throughput method. Thus, the current pilot proposal seeks to accomplish following tasks: a) to design cloning strategies for the construction of a library of the apoptotic reporter cell lines using fluorescent proteins as reporters, and to verify the cloning methods for four representatives of apoptotic regulatory molecules in lung and liver cancer cells; b) to establish 3D cell culture methods using self-assembling nano-scaffold hydrogels in microfluidic channels, and develop a prototype device for a microfluidic 3D living cell array; c) to evaluate our prototype 3D apoptosis microfluidic arrays using natural and synthetic anticancer drugs with known anti-cancer mechanisms, and to develop a data mining strategy. The completion of this pilot project will lead to a joint full proposal to develop a highthroughput functional platform to unveil insight dynamic information in the complex apoptotic signaling network for anti-cancer drug screening and mechanism study. We expect that the system will be eventually used to identify and study novel anti-cancer molecules for efficient and safe cancer treatments or preventions.
H. Gandhok, Z. Dereli-Korkut, X. Jiang and S. Wang, “Real time detection of the dynamic TRAIL-DR4/DR5 interactions using FRET in microfabricated channels”, 2009 BMES Annual Meeting, Pittsburgh, PA, October 2009