Abstract
Overexposure to endocrine disruptor chemicals (EDCs) can result in serious health problems, yet they are commonly found in everyday items such as pesticides, personal care products, nutritional supplements, and plastics. The U.S. Environmental Protection Agency, along with other such agencies from around the world, have therefore mandated that new approaches be designed to screen these products for the presence of EDCs. However, despite the presence of several types of extant EDC detection assays, there still exists a backlog approaching 87,000 chemicals currently awaiting screening. Autobioluminescent detection systems, which utilize cellular bioreporters capable of autonomously modulating bioluminescent signals without the need for external stimulation or investigator interaction, provide an attractive means for addressing this backlog because of their reduced performance costs and increased throughput relative to alternative assay systems. This chapter reviews the variety of existing EDC detection assays and evaluates the performance of a representative autobioluminescent estrogen-responsive EDC bioreporter to provide an overview of how autobioluminescence can be used to improve EDC detection using in vitro assay systems.
Highlights
The human endocrine system is an interconnected, finely tuned network of glands that produce hormones responsible for health and well-being from the time of conception until death
Capitalizing on the difficulties posed by this situation, a multitude of companies have formed to evaluate how the compounds that make up everyday items such as pesticides, personal care products, nutritional supplements, and plastics can imbalance the delicate regulation of normal endocrine function in humans and wildlife
While this work screened the performance of the HEK293-based estrogen-responsive bioreporter against a variety of endocrine disruptors (EDCs) and associated controls, it will be necessary to validate the performance of this assay format at the levels of scale required for commercial use
Summary
The human endocrine system is an interconnected, finely tuned network of glands that produce hormones responsible for health and well-being from the time of conception until death. Autobioluminescence, with is defined as the ability to self-initiate the production of a luminescent signal using only endogenously supplied substrates to perform the enzymatic reactions necessary for signal generation [12], can reduce the number of required assay steps, eliminate the need for superfluous reagent costs, maintain human bioavailability relevance through the use of human cellular hosts, and increase throughput by minimizing hands-on performance time and employing automated processing and detection systems [13]. These benefits are made possible by the autonomous functionality of the synthetic luciferase gene cassette (lux) that controls the autobioluminescent phenotype. Using the detection of estrogenic compounds as an example, this chapter will provide an overview of how these autobioluminescent cellular bioreporters function in this role relative to alternative, traditional Tier 1 EDC sensor platforms and the advantages and disadvantages they provide for addressing the needs of the EDSP21 program
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