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测定法，以分类皮肤敏化剂，该测定法可以在人类皮肤的三维模型的高度相关背景下评估化学物质，以更紧密地模拟该化学物质体内环境。过敏性接触性皮炎（ACD）是一个广泛传播的公共健康问题，该问题是由于人类皮肤暴露于工作场所和家庭中某些反应性化学物质而引起的。确定哪种化学物质诱导ACD对于减少人类暴露至关重要。目前尚无对皮肤敏化的验证测试，这些测试排除了动物的使用，但令人信服的经济和道德原因支持开发新的体外方法，这些方法可以准确预测化学物质或混合物的敏感性，并消除了对动物的需求。 拟议的Stratascreensens 96-Well Assay背后的关键技术创新是一种独特的三维人体皮肤模型，它利用荧光素酶活性作为快速，容易，准确的定量测定读数，以实现皮肤敏感的潜力。在人体皮肤模型的背景下，对皮肤敏化化学物质的准确反应的体外测定1）允许将测试化学物质应用于皮肤表面，从而模仿ACD中实际正常的暴露途径； 2）对体内生理环境进行建模比目前使用的二维单层培养系统更好； 3）使用96井格式来增强筛选吞吐量； 4）减少或替代对皮肤敏化剂评估中动物测试的需求。 Stratascreensens的优势直接解决了21世纪毒理学面临的挑战和局限性，并谈到了美国国家环境健康科学研究所的使命。 该阶段I SBIR提案的目标是生成和表征独特的蜂窝试剂，用于生产Stratascreensens分析中使用的记者皮肤模型。这将通过使用Stratatech的平台技术，NIKS细胞来实现，NIKS细胞是非肿瘤，无病原体人角质形成细胞祖细胞的一致来源。遗传培训NIKS角质形成细胞并产生产生外源性蛋白质的三维皮肤组织的能力为开发对皮肤敏化化学物质响应的记者皮肤模型提供了独特的机会。成功完成I阶段目标将证明我们方法的可行性，并生成Stratascreensens分析中使用的细胞试剂。随后的研究将完善测定参数，支持对测定的验证，这将降低公众接触有害化学物质的风险并减少对动物测试的依赖。 公共卫生相关性：过敏性接触性皮炎（ACD）是一个广泛的公共卫生问题，该问题是由于人类皮肤暴露于工作场所和家庭中某些反应性化学物质而引起的。诱导ACD的化学物质的鉴定对于减少人类暴露至关重要。同样，出于经济和道德原因，消除需要动物的测试很重要。开发一种基于动物的测试，该测试通过减少对动物测试的依赖并降低公众暴露于有害化学物质的风险，从而满足了对药品，消费产品和化学工业的主要需求。