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.项目摘要/摘要 口腔粘膜炎是癌症治疗的一种痛苦的、影响治疗的副作用，影响大约250,000例癌症 在美国每年都有病人。目前，FDA批准的唯一治疗口腔粘膜炎的药物，帕利福明， 仅占临床病例的4%左右。这个项目的长期目标是阐明分子 粘膜炎的损伤起始机制，包括RNA表达特征，以及细胞- 此外，还有助于发现和翻译新的抗粘膜炎候选药物。这 该项目预计将直接支持未来的个性化医学方法，用于治疗口腔粘膜炎 癌症患者。粘膜炎的启动被认为涉及到先天免疫和损伤反应 结构性牙龈细胞，始于上皮下层内皮细胞和间充质细胞，并向 对牙周角质形成细胞和上皮溃烂的损害。因此，开发一种芯片上的牙床， 包括与粘膜炎的启动有关的细胞培养，预计将支持这些目标。 这项提议旨在(1)在暴露于癌症治疗的芯片上确定牙龈破裂的开始，以及 (2)测定牙周膜对治疗口腔粘膜炎药物的反应。取得的成就 目标1将涉及在三种培养芯片中建立细胞功能和分子表达的基线，包括 人牙周角质形成细胞、成纤维细胞和微血管内皮细胞系。生存、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)荧光检测、四唑盐和细胞因子抗体阵列检测 营养培养基及全转录组RNA测序。对四种不同剂量化疗的反应 涉及口腔粘膜炎的药物将在芯片和井板中进行评估并与基线进行比较 使用相同的分析方法进行单一培养。实现目标2将涉及四种抗粘膜炎疗法的测试 在芯片上，对于暴露在一种化疗药物剂量下的培养物，发现会引发粘膜炎的特征， 使用与目标1相同的分析方法。将在所有实验中通过跟踪评估牙龈屏障功能 细胞层上已知大小的荧光标记口腔细菌和碳水化合物分子。 这项提议与健康的相关性在于直接影响口腔健康和支持发展 通过了解口腔粘膜炎和芯片上的治疗效果，实现精确医学方法的研究。 建议的系统将作为人类牙龈粘膜炎的启动模型，并可以测试新的 抗粘膜炎药物候选与临床前试验平行进行。这样的知识可能会导致更多 针对口腔粘膜炎干预措施的临床试验取得成功。翻译成的主要文化 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 通过将癌症治疗与芯片上确定的最好的抗粘膜炎治疗相结合，可以减少粘膜炎的风险。