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

基本信息

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
项目类别：
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

项目摘要

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（由申请人提供）：在人类细胞系中表达细菌生物发光：设计自体发光报告细胞以筛选内分泌干扰物化学物质项目摘要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，以改进对显示内分泌干扰物的化学物质的筛选扰乱活动，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、杀虫剂、药品和个人护理产品的内分泌干扰物化学品 (EDC) 受到密切关注，因为它们会对人类和野生动物的健康产生不利影响，包括癌症发病率增加、精子数量减少、女性性早熟、以及生殖器官功能的改变。在美国 EPA 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效应。由于有 87,000 种化学品可能需要测试，Tier 1 EDC 筛选测定电池已被认为太慢且成本太高，无法满足当前的需求，EPA 的任务是将其 EDSP 计划转向更快、更具成本效益的筛选技术。为了满足这一需求，490 BioTech 提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构建体，以改善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构建体，该基因构建体必须提供底物来激活其光发射响应，从而导致潜在毒理学相互作用的单时间点快照的信息量很小。我们提出的无底物、自生物发光报告细胞系将响应内分泌活性化合物，连续、实时地发光，从而在报告细胞的整个生命周期内，当它与目标化学暴露相互作用和反应时，提供不间断的视觉数据流。此外，使用人体细胞作为传感平台可以提供有关生物利用度和化学物质对人类个体健康的真实影响的更准确和真实的信息。超过 500 个合同实验室对 87,00 种潜在化学品库存进行一级筛选测定，平均成本为 1,000 美元/次，并且 EPA 要求将更快/更好/更便宜的技术整合到一级筛选方案中，我们相信我们拥有的产品能够对化学/药物筛选市场产生重大影响，特别是提高我们对内分泌活性化学物质的理解，因为它们关系到公共健康和消费者安全。