Expressing bacterial bioluminescence in human cell lines: Engineering autobioluminescent reporter cells to screen for endocrine disruptor chemicals Project Summary This Small Business Innovation Research Phase I project proposes to engineer an autobioluminescent human cell line for improved screening of chemicals displaying endocrine disruptor activity to address the National Institute of Environmental Health Sciences (NIEHS) request for tools for improved exposure assessment related to endocrine active chemical exposures. Endocrine disruptor chemicals (EDCs) potentially originating from plastics, pesticides, and pharmaceutical and personal care products are under scrutiny due to implications that they cause adverse health effects in humans and wildlife, including increased cancer rates, lowered sperm counts, early puberty in females, and altered functions of reproductive organs. Chemical manufacturers and importers are being mandated to test their chemicals for endocrine toxicity effects under the auspices of the U.S. EPA Endocrine Disruptor Screening Program (EDSP). With 87,000 chemicals potentially requiring testing, the battery of Tier 1 EDC screening assays have been recognized as being too slow and too costly to meet current demands, and the EPA has been tasked with transitioning its EDSP program towards faster and more cost-effective screening technologies. To address this need, 490 BioTech proposes to engineer a human cell line endowed with a bacterially derived bioluminescent reporter gene construct for improved monitoring of endocrine active chemicals. The current market of bioluminescent reporter cells being applied towards EDC screening rely upon a firefly luciferase gene construct that must be provided with a substrate to activate its light emission response, resulting in only marginally informative single time point snapshots of potential toxicological interactions. Our proposed substrate-free, autobioluminescent reporter cell line will emit light continuously and in real time in response to endocrine active compounds, thus providing an uninterrupted stream of visual data over the lifetime of the reporter cell as it interacts and reacts to target chemical exposure. Further, using a human cell as the sensing platform provides more accurate and realistic information in regards to bioavailability and a chemical's true effect on individual human health. With over 500 contract laboratories performing Tier 1 screening assays at an average cost of $1,000/assay over a potential inventory of 87,00 chemicals, and with the EPA mandating the integration of faster/better/cheaper technologies into the Tier 1 screening scheme, we believe we possess a product capable of significantly impacting the chemical/drug screening market and, here in particular, advancing our understanding of endocrine active chemicals as they pertain to public health and consumer safety.

Public Health Relevance

Certain hormonally active chemicals, collectively referred to as endocrine disruptor chemicals (EDCs), mimic or antagonize the actions of naturally occurring estrogens and are believed to result in adverse developmental, reproductive, neurological, and immune effects in both humans and wildlife. The chemical, pesticide, personal care products, plastics, and nutritional supplements industries have been tasked with screening their products for EDCs. Using a novel autobioluminescent human cell line, 490 BioTech proposes to advance current EDC screening assays with a reporter cell technology that surveys EDC activity continuously and in real time to deliver more data faster and more cost effectively for improved safeguarding of public health.

Agency
National Institute of Health (NIH)
Institute
National Institute of Environmental Health Sciences (NIEHS)
Type
Small Business Innovation Research Grants (SBIR) - Phase I (R43)
Project #
1R43ES022567-01
Application #
8455257
Study Section
Special Emphasis Panel (ZRG1-EMNR-S (10))
Program Officer
Shaughnessy, Daniel
Project Start
2013-02-18
Project End
2014-01-31
Budget Start
2013-02-18
Budget End
2014-01-31
Support Year
1
Fiscal Year
2013
Total Cost
$138,191
Indirect Cost
Name
490 Biotech, Inc.
Department
Type
DUNS #
968832498
City
Knoxville
State
TN
Country
United States
Zip Code
37996
Xu, Tingting; Ripp, Steven; Sayler, Gary S et al. (2014) Expression of a humanized viral 2A-mediated lux operon efficiently generates autonomous bioluminescence in human cells. PLoS One 9:e96347
Xu, Tingting; Close, Dan M; Webb, James D et al. (2013) Autonomously bioluminescent mammalian cells for continuous and real-time monitoring of cytotoxicity. J Vis Exp :e50972