The Center for Alternatives to Animal Testing is an academic center affiliated with the Division of Toxicological Sciences in the Department of Environmental Health Sciences of the Johns Hopkins University Bloomberg School of Public Health.

 

Johns Hopkins School of Public Health

Research Grants 2008-2009

A Rapid Microfluidic Assay for High-Throughput Screening of Chemopreventive Compounds

Pak Kin Wong, PhD
University of Arizona

The overall goal of the project is to develop a rapid screening system for identifying chemopreventive compounds.  Chemopreventive compounds often exert their effects by inducing the expression of cytoprotective enzymes to defend cells from oxidative stress and reactive carcinogenic intermediates.  The expression of many cytoprotective enzymes is regulated by the NF-E2-related factor 2 via antioxidant-responsive elements.  Here, we plan to develop two molecular probe designs for rapid detection of the ARE-bound Nrf2 protein and Nrf2-mediated gene expression for identifying potential chemopreventive compounds.  The molecular probe biosensors will form the technological core of a high-throughput screening system.  To evaluate the assays, human dermal fibroblasts are chosen as an in vitro skin-like model.  The development will lead to a rapid, quantitative, sensitive, and specific screening system for rapid identification of chemopreventive compounds.  The system will provide an alternative for reducing animal usage in predicting the effects of chemopreventive compounds and elucidating their mechanisms of action.  The assays should also be generally applicable to a wide range of toxicology studies, as they are compatible with various in vitro models.