Expressing bacterial bioluminescence in human cell lines: Engineering autobiolumi

描述（由申请人提供）：在人类细胞系中表达细菌生物发光；该小型企业创新研究第一阶段项目建议设计一种自体发光人类细胞系，用于改进对显示内分泌干扰物活性的化学物质的筛选，以满足国家环境健康科学研究所（NIEHS）对改进与内分泌活性化学物质暴露相关的暴露评估工具的要求。可能源自塑料、农药、药品和个人护理产品的内分泌干扰物（EDCs）正受到严格审查，因为它们可能对人类和野生动物的健康造成不利影响，包括癌症发病率增加、精子数量减少、女性青春期提前以及生殖器官功能改变。在美国环境保护署内分泌干扰物筛选计划（EDSP）的支持下，化学品制造商和进口商被要求测试其化学品的内分泌毒性影响。有87,000种化学物质可能需要测试，一级EDC筛选分析方法已经被认为太慢，成本太高，无法满足当前的需求，EPA的任务是将其EDSP计划转变为更快，更具成本效益的筛选技术。为了满足这一需求，490生物技术公司提出设计一种人类细胞系，赋予细菌来源的生物发光报告基因结构，以改善内分泌活性化学物质的监测。目前市场上用于EDC筛选的生物发光报告细胞依赖于萤火虫荧光素酶基因结构，必须提供底物来激活其发光响应，导致潜在毒理学相互作用的单时间点快照只能提供少量信息。我们提出的无底物、自体发光报告细胞系将

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 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.