Phototoxicity Screening Assay in Reconstituted Skin

重建皮肤的光毒性筛选试验

• 批准号: 7113843
• 负责人: George L. DeGeorge
• 金额: $ 66.21万
• 依托单位: MB RESEARCH LABORATORIES, INC.
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-09-01 至 2009-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7113843
• 关键词: artificial skin; biotechnology; clinical research; cytotoxicity; flow cytometry; high throughput technology; microarray technology; skin irritation /irritant; technology /technique development; ultraviolet radiation

DESCRIPTION (provided by applicant): We have developed and prevalidated an in vitro screening test for phototoxicity, the Enhanced Phototoxicity Assay in Reconstituted Skin (EPARS), to replace the current standard animal phototoxicity tests. The phototoxic potential of chemicals, cosmetics, dietary supplements and Pharmaceuticals are a major and growing concern in the consumer products industry, but society and the government have demanded decreased use of experimental animals. EPARS is more objective, faster, and less expensive than animal tests. To-date, no alternative phototoxicity test has been validated in the U.S. (via ICCVAM), although the 3T3 Neutral Red Uptake (3T3NRU) assay has been accepted in the E.U. after validation by ECVAM. EPARS is significantly superior to the 3T3NRU viability test because it overcomes several of the 3T3NRU test's limitations. Specifically: (1) EPARS is based upon a differentiated tissue model that closely parallels human skin morphology, instead of a fibroblast monolayer; (2) the tissues are composed of primary human keratinocytes in a 3D culture system, a more relevant model than a mouse tumor cell line; and (3) test substances can be applied directly, avoiding the often problematic solubilization of formulations into culture media. If developed to its full potential, EPARS would be the most accurate assay available to identify potential phototoxic agents in humans. We propose to confirm the reproducibility, sensitivity, specificity and predictability of EPARS by testing an expanded list of known phototoxins and validate the most predictive and accurate tissue viability endpoints identified in Phase I. We will also use human gene microarrays to measure changes in gene expression patterns and identify genes that can be used as predictors for induction of phototoxicity. This Phase II work will supplement and strengthen the foundation of EPARS in preparation for submission to U.S. regulatory agencies as an alternative to animal phototoxicity testing.

描述（由申请人提供）：我们已经开发了并在重构皮肤（EPARS）中增强的光毒性测定（EPARS）进行了体外筛查测试，以取代当前的标准动物光毒性测试。化学物质，化妆品，饮食补充剂和药品的光毒潜力在消费产品行业中越来越关注，但是社会和政府要求减少对实验动物的使用。 EPAR比动物测试更客观，更快，更便宜。迄今为止，尽管在E.U.经过ECVAM验证后。 EPAR高于3T3NRU生存能力测试，因为它克服了3T3NRU测试的几个局限性。具体而言：（1）EPAR基于一个分化的组织模型，该模型与人体皮肤的形态紧密相似，而不是成纤维细胞单层； （2）组织由3D培养系统中的原代人角质形成细胞组成，这是比小鼠肿瘤细胞系更相关的模型； （3）可以直接应用测试物质，以避免配方经常有问题的溶解化为培养基。如果发育具有全部潜力，EPAR将是最准确的测定法，以鉴定人类潜在的光毒剂。我们建议通过测试已知的光毒素的扩展列表，并验证阶段I中确定的最预测性和准确的组织生存能力终点，以确认EPAR的可重复性，敏感性，特异性和可预测性，我们还将使用人类基因微阵列来测量基因表达模式和基因的变化，这些基因可用于预测光毒性。这项第二阶段的工作将补充和加强EPAR的基础，以准备向美国监管机构提交，以替代动物光毒性测试。