Citation:

TOTH, G. P., I. KNOEBL, A. D. BIALES, D. L. LATTIER, D. C. BENCIC, AND J. M. LAZORCHAK. APPLICATION OF GENOMIC AND PROTEOMIC INDICATORS TO CHARACTERIZE EXPOSURE OF AQUATIC ORGANISMS TO ENVIRONMENTAL CONTAMINANTS. Presented at SETAC, Baltimore, MD, November 13 - 17, 2005.

Impact/Purpose:

The indeterminate condition of exposure indicator research stands to change markedly with the ability to connect molecular biological technologies with cellular or tissue effects and outcomes. Three focal areas of ecological research aim to develop a sequence of approaches where "the earliest recognizable signatures of exposure" (i.e., unique patterns of up- and down-regulated genes and proteins) are identified for numerous stressors, demonstrable in case studies and incorporated into Agency, State and Regional studies supported by EMAP and other programs.

Area 1, Computational Toxicology Research: Exposure assessment has historically been based on use of chemical analysis data to generate exposure models. While biological activity of chemicals has been recognized to be important for exposure risk assessments, measurement of such activity has been limited to whole organism toxicity tests. Use of molecular approaches will:

improve extrapolation between components of source-to-outcome continuum (source , exposure , dose , effect , outcome)

Using a systems modeling approach, gene and protein expression data, in small fish models (fathead minnow and zebrafish), will be integrated with metabolomic and histopathological data. This will assist in prediction of environmental transformation and chemical effects based on structural characteristics, and enhance quantitative risk assessments, including areas of uncertainty such as a basis for extrapolation of effects of endocrine disrupting chemicals, interspecies extrapolation, complex chemical mixtures and dose-response assessment.

Area 2, Ecological Research-Environmental Diagnostics: Development of molecular diagnostic indicators contributes to several of the GPRA Diagnostic Research Goals. Methods will employ DNA microarray technology and expression proteomics, focusing on species of relevance to aquatic ecosystem risk assessment. Significantly, these diagnostic indicators will open the door to understanding subcellular interactions resulting from exposure to complex chemical mixtures.

define relationship between genetic disposition of populations and degree/specificity of stressor-specific gene transcriptional response in aquatic organisms (fish and invertebrates)

identify of chemical mixture induced transcriptional "patterns" using microarrays and hyperspectral scanning - via collaboration with DOE Sandia National Labs

apply molecular indicators to watershed level stressor study, including pilot studies with targeted pesticides and toxins indicators

develop molecular indicators of exposure for invertebrates (Daphnia, Lumbriculus, Chironomus)

Area 3, Exposure Research in Endocrine Disruptors:

Subobjective 1: Develop exposure methods, measurement protocols, and models for assessment of risk management practices of endocrine disrupting compounds. As risk management approaches are identified and developed, there will be a need to identify, adapt and develop bioassay screening tools and other analytical methods to assess their efficacy. Measurements research will be performed to define management needs. This effort will entail cross-lab participation from NRMRL, NERL and NHEERL.

Subobjective 2: Determine extent of environmental and human exposures to EDCs, characterize sources and factors influencing these exposures, develop and evaluate risk management strategies to reduce exposures. In order to develop effective risk management strategies, it is important to understand the extent of exposures to endocrine disrupting compounds and factors influencing source-to-exposure-to-dose relationships.

apply molecular indicators of exposure to estrogenic compounds in selected wastewater treatment plants located in ten USEPA Regions

identify differential gene expression following exposure of fathead minnows to environmental androgens and androgen-like compounds

apply molecular indicators of exposu

Description:

Advances in molecular biological methods are continually being brought to bear on human health research, from a basic understanding of systems biology to identification of toxicity pathways for environmental stressors and to correlations of molecular indicators with physiological outcomes in human epidemiology studies. This prodigious research effort is beginning to be mirrored for aquatic organisms in the academic, government and private sector research laboratories. The U.S. EPA's National Exposure Research Laboratory (NERL) is partnering with other laboratories within E.P.A. as well as the DOE's Sandia National Laboratories and Joint Genome Institute to develop microarray resources for the fathead minnow. Combined with other gene discovery tools and proteomic analyses, NERL scientists are developing molecular methods to diagnose exposure to chemicals and chemical families, such as endocrine-disrupting chemicals (EDCs), pesticides and pharmaceuticals. These tools will be applied to answer real world questions about the extent and level of exposure to and effects on organisms in aquatic ecosystems. Current applications include analyses of estrogenic activity in a national scale study on wastewater treatment plant effluents, partnership with USGS in studying exposure issues related to the finding of intersex in smallmouth bass in the Potomac watershed and analyses of estrogenic activity in several cyprinids sampled during studies as part of the Environmental Monitoring and Assessment Program (EMAP) in the Ohio River watershed. Ultimately, a wide range of molecular indicators will be developed that are phenotypically-anchored to relevant ecological function (e.g., reproduction) and that will allow for characterization of exposure and effects before they are manifested at the tissue, organ, individual, or population level. Data for several projects in which these tools are used to diagnose exposure of aquatic organisms to EDCs in the field will be presented.

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