CAREER: Mechanistic Toxicity Assessment of Emerging Pollutants via Prokaryotic Real-Time Gene Expression Profiling for Water Quality Monitoring

与我们的水中过多的新污染物有关的健康风险和有害环境影响需要在毒性评估技术上取得突破，因为现有的方法既不可行也不足以提供消除这些威胁的监管决策所需的及时信息。这一职业项目的目标是(1)应用创新的跨学科方法的原核生物实时基因表达谱作为评估毒性影响和确定新出现污染物的毒性机制的手段；(2)开发一个更复杂、信息更丰富、但成本效益更高且可行的基因组评估系统，以监测和量化水样中污染物的毒性影响；(3)建立一个创造性的综合教育计划，以吸引和教育来自不同学科和背景的人员，特别是妇女和代表性不足的群体，成为环境工程领域的贡献者。长期目标是建立一个独特的跨学科和转化性研究和教育计划，专门应用基因组和分子生物技术改善和监测水质。缺乏足够和可行的毒性评估和量化方法，极大地阻碍了制定和实施有效的法规、战略和技术，以控制和消除新出现的令人关切的污染物的有害影响。该项目将首次应用由大量(~2000)生物发光绿色荧光蛋白转化的大肠杆菌菌株组成的原核全细胞阵列，以获得对污染物的实时基因表达谱。通过高分辨率和高通量地测量暴露在毒素下的生物体的全球分子状态，将能够同时评估毒性效应、了解毒性机制并获得污染物特定的分子指纹(生物标记物)，用于化合物分类和鉴定。这种方法将导致比传统方法更及时和更有信息量的毒性评估结果。这将大大提高基因表达谱用于毒性评估的可行性和成本效益，因为它的分析程序更简单、更快、更可靠，更高的重用性，以及细胞阵列文库的定制所需的灵活性。此外，与现有的基于基因芯片的基因组图谱方法相比，该方法具有更高的灵敏度和特异性，因为它增加了图谱数据的时间维度，从而允许更全面和准确地评估污染物的毒性。获得大量关于各类新兴环境污染物的毒性效应和毒性机制的新数据，将有助于填补我们对新兴污染物风险的认识的一个巨大空白。将这些信息纳入标准的分级生态风险评估框架，可以减少风险评估的不确定性，优化潜在风险最大的化学品的资源利用，从而大大提高监管生态毒理学的效率。这个跨学科项目将把它的研究产品纳入一个教育计划，该计划建立在跨学科、体验式和多渠道的教育和推广模式基础上，目标是来自不同层次和背景的人员，包括从K-12到研究生水平的学生、水专业人员和从业者、当地机构和监管机构，以及高中和社区大学教师。这项研究将证明基因组毒性评估方法作为传统毒性评估和鉴定测试的替代方法和/或作为常规测试的补充工具的能力和优势。这项研究的结果将对确保水质以保护公众健康和提高生活质量产生重大影响。该项目融合了生物技术、毒理学和环境工程的不同学科，为水质监测的基因组毒性评估研究开辟了新的领域。创造性和综合性的教学和教育模式将增加参与环境工程劳动力的多样性，并使学生掌握足够的知识和技能，以应对当今具有挑战性的环境问题。由国际环保协会首创的BEST(环境生物技术-展示和培训)计划将继续扩大，通过与各种NU教育计划(STEM、FURI、RET和YSP)以及当地组织和社区的联系，增加环境工程领域参与者的多样性，特别是关于妇女、少数族裔和残疾人的参与者。加强技术转让和信息传播将通过ITRI(由PI发起的产业转化研究倡议)、多层次合作和NU合作计划来实现。对私人助理职业发展的支持将帮助她成长为一名成功的学者和教育家，通过促进改善水质和保护公共卫生的技术和教育进步而造福社会。

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.