Expressing bacterial bioluminescence in human cell lines: Engineering autobiolumi

在人类细胞系中表达细菌生物发光：工程自动生物发光

• 批准号: 8455257
• 负责人: Dan Morrison
• 金额: $ 13.82万
• 依托单位: 490 BIOTECH, INC.
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-02-18 至 2014-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8455257
• 关键词: Address; Androgens; Bacterial Luciferases; Behavior; Binding; Biological Assay; Biological Availability; Bioluminescence; Breast Cancer Cell; Cancer cell line; Cell Line; Cells; Characteristics; Chemical Exposure; Chemicals; Complex; Contracts; Data; Development; Endocrine; Endocrine Disruptors; Endocrine system; Engineering; Environmental Health; Environmental Pollution; Equipment and supply inventories; Estradiol; Estrogen Antagonists; Estrogens; European Union; Evaluation; Exposure to; Faslodex(ICI 182,780); Female; Fireflies; Firefly Luciferases; Health; Homeostasis; Hormones; Human; Human Cell Line; Human Engineering; Immune; In Vitro; Individual; Industry; Kinetics; Laboratories; Light; Link; Luc Gene; Maintenance; Malignant Neoplasms; Manufacturer Name; Marketing; Methods; Monitor; National Institute of Environmental Health Sciences; National Toxicology Program; Neurologic; Organ; Output; Performance; Pesticides; Pharmaceutical Cares; Phase; Plastics; Positioning Attribute; Preclinical Drug Evaluation; Puberty; Public Health; Reaction; Reaction Time; Reporter; Reporter Genes; Reproduction; Response Elements; Safety; Scheme; Self Care; Side; Small Business Innovation Research Grant; Specificity; Sperm Count Procedure; Stream; Surveys; T47D; Technology; Testing; Time; Toxic effect; United States Environmental Protection Agency; Visual; consumer product; cost; cost effective; design; dietary supplements; high throughput screening; improved; in vivo; light emission; meetings; novel; programs; public health relevance; reproductive; reproductive function; research and development; response; screening; stable cell line; tool

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.

描述(申请人提供)：在人体细胞系中表达细菌生物发光：设计自体生物发光报告细胞以筛选内分泌干扰物化学物质项目摘要这个小型企业创新研究第一阶段项目建议设计一种自体生物发光人体细胞系，以改进对显示内分泌干扰物活性的化学物质的筛选，以满足美国国家环境健康科学研究所(NIEHS)对与内分泌活性化学物质暴露相关的改进暴露评估工具的要求。可能来自塑料、杀虫剂以及医药和个人护理产品的内分泌干扰物(EDCs)正在接受审查，因为它们可能会对人类和野生动物造成不利的健康影响，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。在美国环保局内分泌干扰物筛选计划(EDSP)的主持下，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效应。由于可能需要检测87,000种化学物质，Tier 1 EDC筛查分析的电池被认为速度太慢，成本太高，无法满足当前的需求，EPA的任务是将其EDSP计划过渡到更快、更具成本效益的筛查技术。为了满足这一需求，490生物技术公司建议设计一种人类细胞系，赋予其细菌来源的生物发光报告基因结构，以改进对内分泌活性化学物质的监测。目前应用于EDC筛选的生物发光报告细胞市场依赖于萤火虫荧光素酶基因结构，该基因结构必须提供底物以激活其光发射响应，导致仅产生潜在毒理学相互作用的微不足道的单一时间点快照。我们提出的无底物、自体生物发光报告细胞系将 响应内分泌活性化合物连续和实时地发光，从而在报告细胞相互作用和对目标化学暴露做出反应时，在报告细胞的整个生命周期内提供不间断的可视数据流。此外，使用人体细胞作为传感平台，可以提供关于生物利用度和化学物质对个人健康的真实影响的更准确和现实的信息。超过500家合同制实验室在87，00种化学品的潜在库存中以每项平均1,000美元的价格进行一级筛查分析，加上美国环保局授权将更快/更好/更便宜的技术整合到一级筛查计划中，我们相信我们拥有一种能够显著影响化学品/药物筛查市场的产品，尤其是在这里，增进我们对内分泌活性化学品的理解，因为它们与公共健康和消费者安全有关。