Development of the UValidate platform for the profiling of topically applied chemical agents.

开发 UValidate 平台，用于分析局部应用的化学制剂。

• 批准号: 10484288
• 负责人: DEAN ROSENTHAL
• 金额: $ 80.92万
• 依托单位: AMELIA TECHNOLOGIES, LLC
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-20 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10484288
• 关键词: 3-Dimensional; Accounting; Address; Aging; Algorithms; Animals; Behavioral; Biological; Biological Assay; Biology; Blood; CRISPR/Cas technology; Catalogs; Cell Line; Cell model; Cells; Chemical Agents; Chemicals; Clustered Regularly Interspaced Short Palindromic Repeats; Complement; Complex; Computer software; Contract Services; Cosmetics; Country; Coupled; Cultured Cells; Cutaneous; DNA Damage; DNA Repair; DNA lesion; Data Storage and Retrieval; Dermal; Development; Diffusion; Digestion; Effectiveness; Ensure; Environment; Exposure to; FDA approved; Face; Fibroblasts; Formulation; Funding; Future; Health; Health Knowledge Attitudes Practice; Human; Individual; Industry; Kinetics; Knock-in; Label; Lead; Lesion; Malignant Neoplasms; Measurement; Measures; Minerals; Modeling; Molecular; Mutagenicity Tests; Mutation; Organ; Patients; Pharmaceutical Preparations; Phase; Poison; Population; Protective Clothing; Quality Control; Reactive Oxygen Species; Reagent; Reproducibility; Research Personnel; Risk; Safety; Services; Short Waves; Skin; Skin Aging; Skin Cancer; Skin Care; Skin tanning; Small Business Technology Transfer Research; Spectrometry; Sunscreening Agents; Surface; Technology; Testing; The Sun; Time; Topical agent; Topical application; UVA induced; Ultraviolet B Radiation; United States; Work; Zinc Oxide; alkalinity; base; cancer initiation; carcinogenesis; carcinogenicity; cell type; combinatorial; data acquisition; data pipeline; deep learning; design; drug discovery; high throughput screening; improved; individual response; irradiation; keratinocyte; melanocyte; novel; prevent; repaired; research clinical testing; screening; skin barrier; skin disorder; solar ultraviolet radiation; success; sun damage; three dimensional cell culture; titanium dioxide; ultraviolet irradiation

Project Summary Skin is the first line of defense against the outside environment. When the skin barrier is breached by UV irradiation (UVR) or toxic chemicals, living cell layers including epidermal keratinocytes, melanocytes and dermal fibroblasts are subjected to DNA damage. If unrepaired, this damage can lead to photoaging and cutaneous carcinogenesis. In fact, skin cancer is by far the most common malignancy, accounting for more cases of cancer in the US and other countries than all other organs combined. Sunscreens have been used extensively with some success, but newer chemical filters suffer from serious shortcomings that make currently available products undesirable, and can be a risk to human health. Current strategies that examine the effectiveness and safety of organic UV filters need to be reassessed. These new strategies should test the biology of UVR in the presence of sunscreens. Further, they need to be high-throughput (HTP) in order to examine combinations of different chemical blockers that are present in most formulations, work in cultured cells for ease of screening, and be animal-friendly. In Phase I we showed proof-of-principle for a novel UVR HTP screening platform called “UValidate, employing mixed populations of isogenic keratinocytes and melanocytes derived from a single donor, exposed to two-compound combinations of sunscreen active ingredients in the presence and absence of short wave (UVB; 285-320), and long wave solar UVR (UVA1; 340-400 nm). These screens measure reactive oxygen species (ROS), DNA damage (CPDs), and micronuclei and show a complex interplay between solar UV wavelength, sunscreen blockers, and cell types. This highlights the pressing need for our platform, compared to currently-utilized assays that measure only the amount of UV that is blocked by spectrometry. In the proposed Phase II, we plan to test known chemical UV blockers for the cosmetic and skin-care markets as a contracting service, using a panel of donor-derived isogenic keratinocytes, melanocytes and fibroblast. At the completion of Phase II, if funded, UValidate will be offered as a service particularly suited for companies at the early stages of drug discovery, regulatory bodies and established pharma seeking to optimize formulation. Advances in this proposal include fluoro-tagging isogenic patient cell lines, establishing 2D and 3D cell cultures to determine individual responses to solar UV irradiation, utilization of reagents and assays to rapidly determine the types of DNA lesions and their repair, and design of AI software to more accurately determine damage. Control cells, generated using CRISPR technology, will reproduce DNA repair-compromised skin cells and 3D skin equivalents will also be included to ensure experimental rigor and reproducibility. The present crisis due to the lack of non- toxic sunscreens has led to class action lawsuits, banning of sunscreens in growing numbers worldwide, and recall of formulations from the market. Therefore, this technology is timely and needed with great expediency to address the urgent need for the discovery and testing of safe and effective UV blockers to prevent the most common, and often debilitating and deadly cancers that we face.

项目概要 当皮肤屏障被紫外线破坏时，皮肤是抵御外界环境的第一道防线。 辐射（UVR）或有毒化学物质、活细胞层，包括表皮角质形成细胞、黑素细胞和真皮 成纤维细胞受到 DNA 损伤，如果不修复，这种损伤会导致光老化和皮肤。 事实上，皮肤癌是迄今为止最常见的恶性肿瘤，占癌症病例的较多。 在美国和其他国家，防晒霜主要与所有其他器官一起使用。 取得了一些成功，但新型化学过滤器存在严重缺陷，使得目前可用的 产品不受欢迎，并且可能对人类健康构成风险。 有机紫外线过滤剂的安全性需要重新评估，这些新策略应该测试紫外线辐射的生物学特性。 此外，它们需要高通量（HTP）才能检查组合。 大多数配方中存在不同的化学阻断剂，可在培养细胞中发挥作用，以便于筛选， 在第一阶段，我们展示了一种名为 UVR HTP 筛选平台的原理验证。 “UValidate 采用来自单个供体的同基因角质形成细胞和黑素细胞的混合群体， 在存在或不存在短暂暴露的情况下暴露于防晒活性成分的两种化合物组合 波（UVB；285-320）和长波太阳 UVR（UVA1；340-400 nm）这些屏幕可测量活性氧。 物种 (ROS)、DNA 损伤 (CPD) 和微核，并显示出太阳紫外线之间复杂的相互作用 相比之下，这凸显了我们平台的迫切需求。 目前使用的测定法仅测量被光谱法阻挡的紫外线量。 第二阶段，我们计划作为承包商测试化妆品和护肤市场已知的化学紫外线阻滞剂 服务，使用一组来自供体的同基因角质形成细胞、黑素细胞和成纤维细胞。 第二阶段，如果获得资助，UValidate 将作为一项特别适合处于早期阶段的公司的服务提供 药物发现、监管机构和知名制药公司寻求优化配方。 建议包括荧光标记同基因患者细胞系、建立 2D 和 3D 细胞培养物以确定 个人对太阳紫外线照射的反应，利用试剂和测定法来快速确定紫外线的类型 DNA损伤及其修复，以及人工智能软件的设计，以更准确地确定损伤控制细胞， 使用 CRISPR 技术生成，将复制 DNA 修复受损的皮肤细胞和 3D 皮肤等效物 还将包括确保实验的严谨性和可重复性。目前由于缺乏非- 有毒防晒霜引发了集体诉讼，全球范围内越来越多的人禁止使用防晒霜，并且 因此，这项技术是及时且非常必要的。 满足发现和测试安全有效的紫外线阻断剂的迫切需要，以预防最严重的 我们面临的常见且常常使人衰弱且致命的癌症。