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(申请人提供)：该项目的最终目标是开发一种用于皮肤敏感剂分类的创新的体外试验，允许在高度相关的三维人体皮肤模型的背景下评估化学物质，该三维模型更接近于体内环境。过敏性接触性皮炎(ACD)是一种广泛存在的公共卫生问题，由人类皮肤在工作场所和家庭中暴露于某些活性化学物质引起。确定哪些化学物质会导致ACD对于减少人类接触至关重要。目前还没有有效的皮肤致敏测试来阻止动物使用，但令人信服的经济和伦理原因支持开发新的体外方法，准确地预测化学物质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一种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。在人体皮肤模型的背景下，对皮肤致敏化学物质提供准确和强大响应的体外检测将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孔格式来提高筛选吞吐量；以及4)在皮肤致敏物质评估中减少或取代动物试验的需要。StrataScreenSENS的优势直接解决了21世纪毒理学面临的挑战和限制，并向国家环境健康科学研究所的使命发声。这项第一阶段SBIR提案的目标是生成和表征独特的细胞试剂，用于生产StrataScreenSENS分析中使用的报告皮肤模型。这将通过使用Stratech的平台技术NIKS细胞来实现，NIKS细胞是无肿瘤、无病原体的人类角质形成细胞前体细胞的始终如一的来源。对NIKS角质形成细胞进行基因工程并生成可产生外源蛋白的三维皮肤组织的能力，为开发对皮肤致敏化学物质做出反应的报道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方法的可行性，并产生用于StrataScreenSENS分析的细胞试剂。随后的研究将完善化验参数，支持验证一种将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的化验。 公共卫生相关性：过敏性接触性皮炎(ACD)是一种广泛传播的公共卫生问题，由人类皮肤在工作场所和家庭中暴露于某些活性化学物质引起。识别导致ACD的化学物质对于减少人类接触是很重要的。同样，由于经济和伦理原因，取消需要动物进行这种鉴定的测试也很重要。开发一种非基于动物的测试来识别导致ACD的化学物质，将通过减少对动物测试的依赖和降低公众接触有害化学物质的风险来满足制药、消费品和化学工业的主要需求。