IDBR: Microfluidic Platform for Accurate Sampling of Biological Signaling Events

Virginia Polytechnic Institute And State University, Blacksburg VA

Investigators

Abstract

An award is made to Virginia Tech to develop a microfluidic device that will enable real-time monitoring of fast cellular events, to increase the fidelity of in vitro generated phosphor-proteomic data necessary for the elucidation of biological signaling pathways. Reversible protein phosphorylation in response to external stimuli represents a key regulatory mechanism that triggers essential cellular signaling events. Aberrant phosphorylation of Ser, Thr and Tyr is often involved in diseases such as cancer, inflammation, diabetes and metabolic disorders. The time-scale of signaling phosphorylation events is very short, ranging from sec to min. Therefore, if the sampling strategy and frequency of the stimulated cells is not adequate, accurate time-dependent concentration profiles of phosphorylated proteins, as well as global profiling of signaling pathways, cannot be performed. The microfluidic device that will be developed in this research will integrate a series of functional elements that will enable the acquisition of accurate time-dependent protein phosphorylation profiles and unambiguous mass spectrometry detection and localization of phosphorylation sites. Phospho-signaling events will be sampled with less than 1 sec delay from occurrence, from phosphor-proteins present in concentrations as low as ~1 fmol. Large-scale integration of functional elements on the chip will lead to 10-100 fold reduction in sample consumption and costs. The microfluidic device will be used to explore the signaling pathways that control cell cycle progression and cellular innate immune responses to Newcastle disease virus in cancer cells. In a broader context, the microfluidic platform will enable the collection of biological data that could help elucidate the mechanisms of aberrant cell behavior, the properties of dysfunctional signaling cascades and of cross-talk between signaling networks, and the development of effective drug treatments for various diseased cell states. On the computational level, it will help discover and validate novel theoretical principles that describe the rate and characteristics of the signal propagation process. On the engineering level, it will facilitate the development of high-performance instrumentation and biomedical infrastructure. This award will enable students and researchers to become visionary scientists that can further advance the forefront of biological research. The education/outreach program will include on-line and on-site interdisciplinary student training in chemistry, biology and engineering concepts, the development of new laboratory modules for graduate and undergraduate students at VT, summer internships to high-school students and under-represented undergraduate minorities, and enhancement of infrastructure for research and education. A dissemination plan will be put in place to engage faculty and scientists with diverse background in collaborations that can benefit from the newly developed technology. The knowledge generated through this research will be disseminated through publications in scientific journals, conference presentations and workshops, and on-line and news-room articles that raise the scientific literacy of the public. Industrial partners will be identified to promote the commercialization of the device.