Career: Mechanistic Toxicity Assessment Of Emerging Pollutants Via Prokaryotic Real-Time Gene Expression Profiling For Water Quality Monitoring

<p>0953633GuThe health risk and harmful environmental impacts associated with the plethora of emerging contaminants in our water necessitate a breakthrough in toxicity-assessment technology because the available methods are neither feasible nor sufficient to provide the timely information needed for regulatory decision making to eliminate these threats. The objectives of this CAREER project are (1) to apply an innovative interdisciplinary approach of prokaryotic real-time gene-expression profiling as a means of evaluating toxic effects and identify toxicity mechanisms of emerging contaminants; (2) to develop a more sophisticated and informative, yet cost-effective and feasible genomic assessment system for monitoring and quantifying toxicity effects from contaminants in water samples; and (3) to establish a creative and integrated education program to attract and educate personnel from various disciplines and backgrounds, especially women and underrepresented groups, to become contributors in the field of environmental engineering. The long-term goal is to build a unique interdisciplinary and translational research and education program that specializes in applying genomic and molecular biotechnology for water-quality improvement and monitoring. The lack of sufficient and feasible methods of toxicity evaluation and quantification greatly hampers the development and implementation of effective regulations, strategies, and technologies for controlling and eliminating the harmful effects from the emerging contaminants of concern. This project will, for the first time, apply a prokaryotic whole-cell array consisting of a large number (~2000) of bioluminescent GFP-transformed E. coli strains to obtain real-time gene-expression profiling in response to contaminants. The high-resolution and high-throughput measurements of the global molecular status of an organism in exposure to a toxin will allow for simultaneous evaluation of toxic effects, understanding of toxicity mechanisms, and obtaining of pollutant-specific molecular fingerprints (biomarkers) for compound classification and identification. This approach will lead to more timely and more informative toxicity-evaluation results than conventional methods. It will greatly improve the feasibility and cost effectiveness of gene-expression profiling for toxicity assessment as a result of its much simpler, faster, and more reliable assay procedure, higher reusability, and desirable flexibility for customization of the cell-array library. Furthermore, the proposed method has higher sensitivity and specificity than the existing microarray-based genomic profiling approach because it adds a temporal dimension to the profiling data and therefore allows for more comprehensive and accurate toxicity evaluation of pollutants. Obtaining a wealth of new data on the toxicity effects and toxic mechanisms for various categories of emerging environmental pollutants will help to fill a great void in our understanding of the risk of emerging contaminants. Incorporation of this information into the standard tiered ecological-risk-assessment framework can greatly advance the efficiency of regulatory ecotoxicology by reducing the uncertainty in risk assessment and optimizing the resource utilization on chemicals with the greatest potential risk. This cross-disciplinary project will incorporate its research products into an education plan built upon an interdisciplinary, experiential, and multi-channel education and outreach paradigm that will target personnel from various levels and backgrounds, including students from K-12 to graduate level, water professionals and practitioners, local agencies and regulators, and high school and community college teachers. This study will demonstrate the ability and advantages of genomic toxicity-assessment methodology as an alternative to conventional tests of toxicity evaluation and identification and/or as a complementary tool to conventional tests. The results of this research will have a significant impact on ensuring water quality for public health protection and quality-of-life improvement. This project integrates the diverse disciplines of biotechnology, toxicology, and environmental engineering and opens new ground for research in genomic toxicity assessment for water-quality monitoring. The creative and integrated teaching and education paradigm will increase the diversity of participation in the environmental engineering workforce and equip students with adequate knowledge and skills to tackle today's challenging environmental issues. The BEST (Biotechnology for the Environment-Showcase and Training) program that was pioneered by the PI will continue to expand through connections with various NU education programs (STEM, FURI, RET, and YSP) and local organizations and communities to increase the diversity of participants in the field of environmental engineering, especially with regard to women, minorities, and people with disabilities. Enhanced technology transfer and information dissemination will be achieved through ITRI (Industrial Translational Research Initiative, initiated by the PI), multi-level collaborations, and the NU co-op program. The support for the PIs career development will help her mature into a successful scholar and educator who benefits society by contributing to the technological and educational advancement of water-quality improvement and public health protection.</p>