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.

项目摘要 皮肤是针对外部环境的第一道防线。当紫外线破坏皮肤屏障时 辐照（UVR）或有毒化学物质，活细胞层，包括表皮角质形成细胞，黑色素细胞和皮肤 成纤维细胞受到DNA损伤。如果未修复，这种损坏可能导致光泽和皮肤 致癌作用。实际上，皮肤癌是迄今为止最常见的恶性肿瘤 在美国和其他国家，除了所有其他器官合并。防晒霜已被广泛使用 一些成功，但是较新的化学过滤器遭受了严重的缺点，这使当前可用 产品不受欢迎，可能是人类健康的风险。当前检查有效性和 有机紫外线过滤器的安全性需要重新评估。这些新策略应测试UVR的生物学 存在防晒霜。此外，它们需要是高通量（HTP），以检查 大多数公式中存在的不同化学阻滞剂，在培养细胞中起作用，以便于筛选， 并对动物友好。在第一阶段，我们显示了一个新型UVR HTP筛选平台的原理证明 “紫瓦比则采用了源自单个供体的异源角质形成细胞和黑素细胞的混合种群， 在存在和不存在的情况下，暴露于防晒活性成分的两体化合物组合 波（UVB; 285-320）和长波太阳UVR（UVA1; 340-400 nm）。这些筛选测量活性氧 物种（ROS），DNA损伤（CPD）和微核，并在太阳紫外线之间显示复杂的相互作用 波长，防晒霜和细胞类型。这突出了我们平台的紧迫需求， 当前利用的测定法仅测量被光谱阻断的紫外线量。在提议中 第二阶段，我们计划测试已知的化学紫外线阻滞剂，以作为合同的化妆品和皮肤护理市场 服务，使用供体衍生的等生角质形成细胞，黑色素细胞和成纤维细胞的面板。完成 第二阶段（如果资助）将作为一项特别适合的服务，特别适合于 药物发现，监管机构和建立的药物，以优化配方。进步 提案包括氟氟的等生患者细胞系，建立2D和3D细胞培养物以确定 个人对太阳紫外线辐照的反应，利用试剂和测定法以迅速确定的类型 DNA病变及其修复以及AI软件的设计，以更准确地确定损害。控制细胞， 使用CRISPR技术生成，将重现DNA修复型皮肤细胞和3D皮肤等效物 还将包括在内，以确保实验性严格和可重复性。由于缺乏非 - 有毒的防晒霜导致了集体诉讼，禁止全球越来越多的防晒霜，并且 召回市场公式。因此，这项技术是及时的，需要很大的权宜 迫切需要发现和测试安全有效的紫外线阻滞剂，以防止最大 我们面临的常见，经常使人衰弱和致命的癌症。