A powerful, fast experimental approach to screen potential BPA substitutes

Earlier, Mancini and his colleagues developed a new, powerful experimental approach that combined high throughput microscopy techniques with specifically engineered cell lines and roboticized screening resources to individually test the effect of numerous compounds on biological functions; these experiments are highly efficient, generating a large number mechanistic and phenotypic measurements simultaneously, even with only very brief exposure of compounds to cells.

We previously established highly multiplexed, single cell-oriented model systems to identify mechanisms involved in complex hormonal biology, Mancini said. Unlike standard biochemical or toxicological assays, our approach quantifies levels of estrogen receptors, nuclear localization, DNA binding, large-scale chromatin modeling, protein interactions and transcription, and also include data on toxicity, cell proliferation and many other characteristics; all at the level of individual cells and in one assay that only takes a few hours.

A robotic system processes the samples and takes tens of thousands of images of the cells through an automated microscope. Later, in-house developed software analyzed and reported on more than 10 billion data points to create a comprehensive picture of what is going on inside and on the surface of the cells.

When the paper describing our novel approach was published, Valspar Corp., an industrial coatings company, approached our lab, Mancini said. They were interested in finding a compound that does not have activity on estrogen receptors, but still provides a means to extend the shelf life of canned foods. Traditional toxicology studies conducted in animals are time consuming, expensive and provide limited data regarding the mechanism involved. Our approach is much faster, sensitive and accurate as standard biochemical assays and allows for testing for numerous compounds at once.

The researchers used their automated approach to screen a number of BPA substitute candidates for their ability to bind to estrogen receptors and trigger their activity.

Using our high-throughput assay, we identified two compounds that are relatively inactive when compared to the negative effects attributed to BPA or the BPA-substitutes in use today, Szafran said.

The compounds we found passed our testing, but it doesnt mean that they are completely free of effects, Mancini said. This would need further testing in animal studies.

Other contributors to this wok include Fabio Stossi, Maureen G. Mancini and Cheryl L. Walker.

This work was supported by the National Institute of Environmental Health Sciences (NIEHS) grants NIEHS R01 (1R01ES023206-01) and NIEHS P30 (ES023512-01) and the Center of Excellence in Environmental Health. Further support was provided by the Integrated Microscopy Core at Baylor College of Medicine with funding from the John S. Dunn Gulf Coast Consortium for Chemical Genomics, the Dan L. Duncan Cancer Center (National Institutes of Health (NIH) P30CA125123), the NIH grants HD007495, DK56338 and K12DK0083014 and the multidisciplinary K12 Urologic Research Career Development Program. DeepBio, Inc received funds and material support from the Valspar Corporation.

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Researchers identify potentially safer substitutes for BPA - Baylor College of Medicine News (press release)