1 Novartis Institutes for BioMedical Research (NIBR), Developmental and Molecular Pathways (DMP), Basel, Switzerland.
SLAS Discov. 2017 Jun;22(5):571-582. doi: 10.1177/2472555216683651. Epub 2017 Jan 6.
Oral and intestinal mucositis is a debilitating side effect of radiation treatment. A mouse model of radiation-induced mucositis leads to weight loss and tissue damage, reflecting the human ailment as it responds to keratinocyte growth factor (KGF), the standard-of-care treatment. Cultured intestinal crypt organoids allowed the development of an assay monitoring the effect of treatments of intestinal epithelium to radiation-induced damage. This in vitro assay resembles the mouse model as KGF and roof plate-specific spondin-1 (RSPO1) enhanced crypt organoid recovery following radiation. Screening identified compounds that increased the survival of organoids postradiation. Testing of these compounds revealed that the organoids changed their responses over time. Unbiased transcriptome analysis was performed on crypt organoid cultures at various time points in culture to investigate this adaptive behavior. A number of genes and pathways were found to be modulated over time, providing a rationale for the altered sensitivity of the organoid cultures. This report describes an in vitro assay that reflects aspects of human disease. The assay was used to identify bioactive compounds, which served as probes to interrogate the biology of crypt organoids over prolonged culture. The pathways that are changing over time may offer potential targets for treatment of mucositis.
口腔和肠道黏膜炎是放射治疗的一种使人虚弱的副作用。一种放射诱导的黏膜炎的小鼠模型导致体重减轻和组织损伤，反映了人类疾病对角质细胞生长因子 (KGF) 的反应，这是一种标准的治疗方法。培养的肠隐窝类器官允许开发一种检测方法，监测治疗对放射诱导损伤的肠上皮的影响。这种体外检测与小鼠模型相似，因为 KGF 和顶板特异性的 spondin-1 (RSPO1) 增强了隐窝类器官在放射后的恢复。筛选出了一些化合物，这些化合物能增加类器官在放射后的存活率。对这些化合物的测试表明，类器官随着时间的推移改变了它们的反应。对隐窝类器官培养物在不同时间点的转录组进行了无偏分析，以研究这种适应性行为。随着时间的推移，发现了许多基因和途径被调节，为类器官培养物的敏感性改变提供了依据。本报告描述了一种反映人类疾病某些方面的体外检测方法。该检测方法被用来鉴定生物活性化合物，这些化合物作为探针，在长时间的培养过程中，对隐窝类器官的生物学进行了研究。随着时间的推移而变化的途径可能为治疗黏膜炎提供潜在的靶点。
