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，大约影响25万癌症患者。 每年在美国的病人。目前，FDA唯一批准的治疗口腔粘膜炎的药物帕利弗明， 只有4%的临床病例该项目的长期目标是阐明分子 粘膜炎损伤起始的机制，包括RNA表达特征和细胞凋亡的时间， 特定事件，并且还促进新的抗粘膜炎药物候选物的发现和翻译。这 该项目预计将直接支持未来治疗口腔粘膜炎的个性化药物方法， 癌症患者。粘膜炎的发生被认为涉及先天性免疫和损伤反应， 结构牙龈细胞，开始于上皮下内皮细胞和间充质细胞，并向 损伤牙龈角质细胞和上皮溃疡。因此，牙龈芯片的开发， 包括培养与粘膜炎的起始有关的细胞，预期将支持这些目的。 该提案旨在（1）确定暴露于癌症治疗的芯片上牙龈破坏的开始，以及 (2)确定芯片上牙龈对口腔粘膜炎的推定药物治疗的反应。实现 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， 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，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 细胞上的（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。对四种剂量的四种化疗的反应 将评估与口腔粘膜炎有关的药剂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剂的培养物，其剂量被发现会引起粘膜炎的标志， 使用与目标1相同的测定。牙龈屏障功能将在所有实验中通过跟踪 荧光标记的口腔细菌和已知大小的碳水化合物分子穿过细胞层。 这项建议的健康相关性在于直接影响口腔健康和支持发展 通过口腔粘膜炎的知识和芯片治疗的效果， 所提出的系统将作为人类牙龈中粘膜炎起始的模型，并且可以测试新的 抗粘膜炎药物候选物与临床前测试平行。这些知识可能会导致更多 在针对口腔粘膜炎干预的临床试验水平上取得成功。翻译成原代培养 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 通过将癌症治疗与最好的抗粘膜炎治疗配对，在芯片上确定。