Expressing bacterial bioluminescence in human cell lines: Engineering autobioluminescent reporter cells to screen for endocrine disruptor chemicals

DESCRIPTION provided by applicant 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 II project proposes to build upon our successful Phase I demonstration of autonomous endocrine disruptor chemical EDC detection using human cell lines to engineer a novel Tier screening assay for the low cost high throughput detection of estrogenically and androgenically active compounds across multiple human tissue types to address the National Institute of Environmental Health Sciences NIEHS request for Improved Test Systems for Prioritization and Safety Evaluation Current Tier EDC screening approaches require the use of non human cell lines that can obscure bioavailability data employ radioactive materials that require dedicated use areas and specially trained personnel or rely upon the use of expensive analytical equipment that prevents high throughput testing Furthermore following this initial Tier screen those chemicals that putatively express endocrine disrupting activities must still be screened against Tier animal models to validate the results Currently there are over contract testing service companies in the U S alone that perform Tier style assays for the chemical pesticide and personal care products industries at an average cost of $ per assay However as the number of potential EDCs requiring testing expands and with many states now adopting legislation to begin screening all municipal water supplies for EDCs market growth within this sector is expected to increase at an annual rate of to approach $ billion by clearly demonstrating the need for new assays that can reduce screening costs increase screening throughput and provide more relevant human bioavailability data without necessitating the use of animal models The EDC responsive substrate free autobioluminescent reporter cell lines developed by BioTech under our Phase I efforts demonstrated the ability to detect EDCs similarly to existing Tier screening assays while employing a human cell culture based model that significantly reduced the cost and personnel effort of testing while simultaneously providing an uninterrupted stream of visual data over the lifetime of the reporter cell as it interacted with and reacted to EDC treatment In this proposal we will leverage this technology to develop fully self contained human cell based high throughput screening assays that provide more accurate and realistic information in regards to EDC bioavailability and effects on human health With application of these assays reducing the cost of Tier EDC screening over fold to less than $ per compound we believe we possess a product capable of significantly impacting the EDC screening market and advancing our understanding of novel compound endocrine disruptor activity as it pertains to public health and consumer safety PUBLIC HEALTH RELEVANCE In response to public and scientific concerns that chemical exposure can interfere with the delicate balance of the endocrine system to trigger adverse health effects in humans and animals government agencies worldwide have implemented comprehensive screening programs to characterize tens of thousands of compounds for endocrine disruptor activity Unfortunately the complex nature and high cost of the screens currently used for these efforts have thus far only been able to generate data on a limited number of chemicals resulting in an extensive untested backlog that is hindering our understanding of how endocrine disruption impacts human and ecosystem health In this Phase II Randamp D effort BioTech proposes to implement a novel endocrine disruptor screening strategy based upon a `humanizedandapos bacterial luciferase genetic construct that links endocrine disruptor activity to autonomous bioluminescent light emission to provide increased data workflows at a greater speed and with less cost than any existing endocrine disruptor assay