A high throughput, in vitro screening system for treatments of oral mucositis in cancer

用于治疗癌症口腔粘膜炎的高通量体外筛选系统

• 批准号: 10041746
• 负责人: Christopher B Raub
• 金额: $ 14.66万
• 依托单位: CATHOLIC UNIVERSITY OF AMERICA
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-01 至 2022-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10041746
• 关键词: Achievement; Affect; Alprostadil; Anti-Inflammatory Agents; Antibodies; Apoptosis; Bacterial Infections; Biological; Biological Assay; Biopsy; Cancer Patient; Carbohydrates; Cell Culture Techniques; Cell Line; Cell Survival; Cell physiology; Cells; Cellular Morphology; Chemotherapy and/or radiation; Chemotherapy-Oncologic Procedure; Cisplatin; Clinical; Clinical Trials; Conflict (Psychology); Development; Devices; Diffusion; Dose; Dose-Limiting; Doxorubicin; Eating; Endothelial Cells; Endothelium; Epithelial; Epithelium; Etiology; Evaluation; Event; Exposure to; FDA approved; Fibroblasts; Fluorescence; Fluorouracil; Funding; Future; Gene Expression Profile; Genetic Transcription; Gingiva; Goals; Health; Hour; Human; Immune; Impairment; In Situ; In Situ Nick-End Labeling; In Vitro; Interleukin-6; Intervention; Knowledge; Label; Lead; Malignant Neoplasms; Measures; Mesenchymal; Methotrexate; Modeling; Molecular; Monoclonal Antibodies; Mucositis; Mucous Membrane; National Institute of Dental and Craniofacial Research; Neutropenia; Nutrient; Oral; Oral health; Pain; Patient risk; Patients; Pattern; Pharmaceutical Preparations; Pharmacotherapy; Preclinical Testing; Proteomics; Radiation therapy; Sodium Chloride; Stains; Structure; Symptoms; System; Testing; Tetrazolium; Tissue Microarray; Tissues; Translating; Translations; Ulcer; United States; United States National Institutes of Health; Vascular Endothelial Cell; base; cancer therapy; cell type; chemotherapy; cytokine; drug candidate; experimental study; fluorescein isothiocyanate dextran; high risk; high throughput screening; individual response; innovation; keratinocyte; keratinocyte growth factor; novel; oral bacteria; oral mucositis; personalized medicine; pre-clinical; precision medicine; response; screening; side effect; spatiotemporal; success; transcriptome; transcriptome sequencing; treatment effect; treatment strategy

7. Project Summary/Abstract Oral mucositis is a painful, treatment-modifying side effect of cancer therapy that affects roughly 250,000 cancer patients annually in the United States. Currently, the only FDA-approved drug to treat oral mucositis, palifermin, is in only about 4% of clinical cases. The long-term objectives of this project are to elucidate molecular mechanisms of damage initiation in mucositis, including RNA expression signatures, and the timing of cell- specific events, and also to facilitate the discovery and translation of new anti-mucositis drug candidates. This project is expected to directly support a future, personalized medicine approach for treating oral mucositis in cancer patients. The initiation of mucositis is thought to involve the innate immune and damage response of structural gingival cells, beginning with sub-epithelial endothelial and mesenchymal cells, and evolving toward damage to gingival keratinocytes and epithelial ulceration. Therefore, development of a gingiva-on-a-chip, involving culture of the cells implicated in the initiation of mucositis, is expected to support these objectives. This proposal aims to (1) determine initiation of gingival disruption on a chip exposed to cancer therapies, and (2) determine responses of the gingiva-on-a-chip to putative drug treatments for oral mucositis. Achievement of Aim 1 will involve establishing a baseline of cell function and molecular expression in a tri-culture chip featuring human gingival keratinocyte, fibroblast, and microvascular endothelial cell lines. Viability, apoptosis, proliferation, transcriptional programming, and cytokine secretion of each cell type on the chip will be determined using (immuno)fluorescence assays on cells, tetrazolium salt and cytokine antibody array assays on conditioned nutrient media, and whole transcriptome RNA sequencing. Responses to four doses of four chemotherapy agents implicated in oral mucositis will be assessed and compared to baseline in the chip and in wellplate monoculture using the same assays. Achievement of Aim 2 will involve testing of four anti-mucositis therapies on the chip, for cultures exposed to one chemotherapy agent at a dose found to elicit hallmarks of mucositis, using the same assays as in Aim 1. Gingival barrier function will be assessed in all experiments by tracking fluorescently labeled oral bacterium and carbohydrate molecules of known size across the cell layers. The health-relatedness of this proposal lies in directly impacting oral health and supporting development of precision medicine approaches through knowledge of oral mucositis and the effects of treatments on a chip. The proposed system would serve as a model of the initiation of mucositis in human gingiva and could test novel anti-mucositis drug candidates in parallel with pre-clinical tests. Such knowledge would potentially lead to more successes at the level of clinical trials targeting oral mucositis interventions. Translated to primary culture of gingival biopsies of cancer patients, the chip-based platform would support personalized modeling and mitigation of mucositis by pairing cancer therapies with the best anti-mucositis therapy, determined on the chip.

7. 项目总结/摘要 口腔粘膜炎是癌症治疗的一种痛苦的、改变治疗效果的副作用，影响大约 250,000 名癌症患者 美国每年都有患者。目前唯一获得 FDA 批准的治疗口腔粘膜炎的药物是 palifermin， 仅占临床病例的 4% 左右。该项目的长期目标是阐明分子 粘膜炎损伤起始机制，包括 RNA 表达特征和细胞损伤发生的时间 特定事件，同时也促进新的抗粘膜炎候选药物的发现和转化。这 该项目预计将直接支持未来治疗口腔粘膜炎的个性化医疗方法 癌症患者。粘膜炎的发生被认为涉及先天免疫和损伤反应 结构性牙龈细胞，从上皮下内皮细胞和间充质细胞开始，并向 牙龈角化细胞受损和上皮溃疡。因此，开发出牙龈芯片， 涉及培养与粘膜炎发生有关的细胞，预计将支持这些目标。 该提案的目的是（1）确定暴露于癌症治疗的芯片上牙龈破坏的开始，以及 (2) 确定芯片牙龈对口腔粘膜炎的假定药物治疗的反应。成就 目标 1 将涉及在三培养芯片中建立细胞功能和分子表达的基线，该芯片具有 人牙龈角质形成细胞、成纤维细胞和微血管内皮细胞系。活力、细胞凋亡、增殖、 芯片上每种细胞类型的转录编程和细胞因子分泌将使用以下方法确定 细胞（免疫）荧光测定、四唑盐和细胞因子抗体阵列测定 营养培养基和全转录组 RNA 测序。对四种化疗的四种剂量的反应 将评估与口腔粘膜炎有关的药物，并与芯片和孔板中的基线进行比较 使用相同的测定进行单一培养。目标 2 的实现将涉及四种抗粘膜炎疗法的测试 在芯片上，对于暴露于一种化疗药物的培养物，其剂量被发现会引起粘膜炎的特征， 使用与目标 1 相同的测定方法。所有实验中都将通过追踪来评估牙龈屏障功能 荧光标记的口腔细菌和细胞层中已知大小的碳水化合物分子。 该提案与健康的相关性在于直接影响口腔健康并支持发展 通过口腔粘膜炎的知识和芯片上的治疗效果来实现精准医疗方法。 所提出的系统将作为人类牙龈粘膜炎起始的模型，并可以测试新的 抗粘膜炎候选药物与临床前测试同时进行。这些知识可能会带来更多 针对口腔粘膜炎干预措施的临床试验取得了成功。翻译成主流文化 癌症患者的牙龈活检，基于芯片的平台将支持个性化建模和缓解 通过将癌症疗法与芯片上确定的最佳抗粘膜炎疗法配对来治疗粘膜炎。