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）：我们已经开发并预先验证了光毒性体外筛选测试，即重建皮肤增强光毒性测定（EPARS），以取代当前的标准动物光毒性测试。化学品、化妆品、膳食补充剂和药品的潜在光毒性是消费品行业日益关注的一个主要问题，但社会和政府要求减少实验动物的使用。 EPARS 比动物试验更客观、更快、更便宜。迄今为止，尽管欧盟已接受 3T3 中性红摄取 (3T3NRU) 检测，但美国尚未验证替代光毒性测试（通过 ICCVAM）。经过ECVAM验证后。 EPARS 明显优于 3T3NRU 活力测试，因为它克服了 3T3NRU 测试的一些限制。具体来说：（1）EPARS基于与人类皮肤形态非常相似的分化组织模型，而不是基于成纤维细胞单层； (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) 测试物质可以直接应用，避免了制剂在培养基中溶解时经常出现的问题。如果充分发挥其潜力，EPARS 将成为识别人类潜在光毒性物质的最准确的检测方法。我们建议通过测试扩大的已知光毒素列表来确认 EPARS 的再现性、敏感性、特异性和可预测性，并验证第一阶段确定的最具预测性和准确的组织活力终点。我们还将使用人类基因微阵列来测量基因表达的变化模式并识别可用作诱导光毒性预测因子的基因。这项第二阶段的工作将补充和加强 EPARS 的基础，为提交给美国监管机构作为动物光毒性测试的替代方案做好准备。