StrataScreenSENS, Skin Sensitizer Detection in Luminescent 3D Human Skin Models

StrataScreenSENS，发光 3D 人体皮肤模型中的皮肤致敏剂检测

• 批准号: 8393318
• 负责人: Cathy Rasmussen
• 金额: $ 20万
• 依托单位: STRATATECH CORPORATION
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-07-27 至 2014-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8393318
• 关键词: Address; Adherent Culture; Allergic Contact Dermatitis; Animal Testing; Animals; Biological Assay; Cell Line; Cells; Characteristics; Chemical Industry; Chemicals; Classification; Clinical; Clone Cells; Coupled; Cytokine Signaling; Detection; Development; Economics; Engineering; Environment; Ethics; Evaluation; Exposure to; Genes; Goals; Home environment; Human; Hypersensitivity skin testing; In Vitro; Industry; Interleukin-18; International Agencies; Journals; Luciferases; Metabolism; Methods; Mission; Modeling; National Institute of Environmental Health Sciences; Nature; Pharmacologic Substance; Phase; Physiological; Production; Proteins; Public Health; Reagent; Reliance; Reporter; Research; Risk; Route; Screening procedure; Skin; Skin Tissue; Small Business Innovation Research Grant; Source; Surface; System; TSLP gene; Technology; Testing; Therapeutic Uses; Thick; Toxicology; Validation; Workplace; base; commercialization; consumer product; experience; exposed human population; in vitro Assay; in vitro testing; in vivo; in vivo Model; innovation; keratinocyte; luminescence; pathogen; predictive modeling; progenitor; promoter; response; technological innovation; three-dimensional modeling; two-dimensional

DESCRIPTION (provided by applicant): The ultimate goal of this project is to develop an innovative in vitro assay for the classification of skin sensitizers that permits evaluation of chemicals within the highly relevant context of a three-dimensional model of human skin that more closely mimics the in vivo environment. Allergic contact dermatitis (ACD) is a wide- spread public health problem resulting from exposure of human skin to certain reactive chemicals in the workplace and in the home. Identifying which chemicals induce ACD is critical to reduce human exposure. There are currently no validated tests for skin sensitization that preclude animal use, yet compelling economic and ethical reasons support development of new in vitro methods that accurately predict the sensitizing potential of a chemical or mixture and eliminate the need for animals. The key technological innovation behind the proposed StrataScreenSENS 96-well assay is a unique three- dimensional human skin model that utilizes luciferase activity as a rapid, easy and accurate quantitative assay readout for skin sensitizing potential. An in vitro assay that provides accurate and robust responses to skin sensitizing chemicals in the context of a human skin model would 1) permit the application of test chemicals to a skin surface, mimicking the actual normal route of exposure in ACD; 2) model the in vivo physiological environment better than the two-dimensional, monolayer culture systems currently employed; 3) utilize a 96- well format to enhance screening throughput; and 4) reduce or replace the need for animal testing in skin sensitizer evaluation. The advantages of StrataScreenSENS directly address the challenges and limitations facing toxicology in the 21st century and speak to the Mission of the National Institute of Environmental Health Sciences. The goals of this Phase I SBIR proposal are to generate and characterize unique cellular reagents for production of the reporter skin model utilized in the StrataScreenSENS assay. This will be accomplished through use of Stratatech's platform technology, the NIKS cells, a consistent source of non-tumorigenic, pathogen-free human keratinocyte progenitors. The ability to genetically engineer NIKS keratinocytes and generate three- dimensional skin tissue that produces exogenous protein provides a unique opportunity to develop reporter skin models responsive to skin sensitizing chemicals. Successful completion of the Phase I Aims will demonstrate the feasibility of our approach and generate the cellular reagents employed in the StrataScreenSENS assay. Subsequent studies will refine assay parameters, supporting validation of an assay that will reduce the risk of public exposure to harmful chemicals and decrease the reliance on animal testing. PUBLIC HEALTH RELEVANCE: Allergic contact dermatitis (ACD) is a wide-spread public health problem resulting from exposure of human skin to certain reactive chemicals in the workplace and in the home. The identification of chemicals which induce ACD is important to reduce human exposure. Similarly, the elimination of tests requiring animals for this identification is important for economic and ethical reasons. Development of a non-animal-based test that identifies chemicals which induce ACD will fill a major need for the pharmaceutical, consumer products, and chemical Industries by decreasing reliance on animal testing and reducing the risk of public exposure to harmful chemicals.

描述（由申请人提供）：该项目的最终目标是开发一种创新的体外检测方法，用于皮肤致敏剂的分类，允许在高度相关的人体皮肤三维模型的背景下评估化学物质，该模型更接近地模拟体内环境。过敏性接触性皮炎（ACD）是一种广泛传播的公共卫生问题，是由于人体皮肤在工作场所和家中接触某些反应性化学物质而引起的。确定哪些化学物质会诱发 ACD 对于减少人类接触至关重要。目前还没有经过验证的皮肤致敏测试可以排除动物使用，但令人信服的经济和伦理原因支持开发新的体外方法，准确预测化学品或混合物的致敏潜力并消除对动物的需求。 拟议的 StrataScreenSENS 96 孔检测背后的关键技术创新是独特的三维人体皮肤模型，该模型利用荧光素酶活性作为皮肤致敏潜力的快速、简单和准确的定量检测读数。在人体皮肤模型中对皮肤致敏化学物质提供准确而稳健的反应的体外测定将 1) 允许将测试化学物质应用于皮肤表面，模拟 ACD 中实际的正常暴露途径； 2）比目前使用的二维单层培养系统更好地模拟体内生理环境； 3) 利用96孔板形式提高筛选通量； 4) 减少或取代皮肤致敏剂评估中动物试验的需要。 StrataScreenSENS 的优势直接解决了 21 世纪毒理学面临的挑战和限制，并体现了国家环境健康科学研究所的使命。 该 I 期 SBIR 提案的目标是生成并表征独特的细胞试剂，用于生产 StrataScreenSENS 测定中使用的报告皮肤模型。这将通过使用 Stratatech 的平台技术 NIKS 细胞来实现，NIKS 细胞是非致瘤、无病原体的人类角质形成细胞祖细胞的一致来源。对 NIKS 角质形成细胞进行基因改造并生成可产生外源蛋白质的三维皮肤组织的能力为开发对皮肤致敏化学物质敏感的报告皮肤模型提供了独特的机会。第一阶段目标的成功完成将证明我们的方法的可行性，并生成 StrataScreenSENS 测定中使用的细胞试剂。后续研究将完善化验参数，支持化验验证，从而降低公众接触有害化学物质的风险，并减少对动物试验的依赖。 公共卫生相关性：过敏性接触性皮炎 (ACD) 是一种广泛存在的公共卫生问题，是由于人体皮肤在工作场所和家中接触某些反应性化学物质而引起的。识别诱发 ACD 的化学物质对于减少人类接触非常重要。同样，出于经济和伦理原因，取消需要动物进行鉴定的测试也很重要。开发一种非动物测试来识别诱发 ACD 的化学物质，将减少对动物测试的依赖并降低公众接触有害化学物质的风险，从而满足制药、消费品和化学工业的主要需求。