Sarvestani Samaneh K, Signs Steven A, Lefebvre Veronique, Mack Stephen, Ni Ying, Morton Andrew, Chan Ernest R, Li Xiaoxia, Fox Paul, Ting Angela, Kalady Matthew F, Cruise Michael, Ashburn Jean, Stiene Jennifer, Lai Wei, Liska David, Xiang Shao, Huang Emina H
Department of Stem Cell Biology and Regenerative Medicine, Lerner Research Institute, Cleveland Clinic, Cleveland, Ohio, USA.
Department of Cell and Molecular Medicine, Cleveland Clinic, Cleveland, Ohio, USA.
Oncotarget. 2018 Jun 19;9(47):28717-28730. doi: 10.18632/oncotarget.25617.
Ulcerative colitis (UC) is a prevalent form of inflammatory bowel disease (IBD) whose pathogenic mechanisms remain unclear. Elucidating these mechanisms is important to reduce UC symptoms and to prevent UC progression into colitis-associated colon cancer (CAC). Our goal was to develop and validate faithful, human-derived, UC models and analyze them at histologic, transcriptomic and epigenetic levels to allow mechanistic studies of UC and CAC pathogenesis. We generated patient-derived primary-organoid cultures from UC and non-IBD colonic epithelium. We phenotyped them histologically and used next-generation-sequencing approaches to profile whole transcriptomes and epigenomes of organoids and primary tissues. Tissue organization and expression of mucin 2 (MUC2) and lysozyme (LYZ) demonstrated histologic faithfulness of organoids to healthy and diseased colonic epithelium. Transcriptomic analyses showed increased expression of inflammatory pathways in UC patient-derived organoids and tissues. Profiling for active enhancers using the H3K27ac histone modification revealed UC-derived organoid enrichment for pathways indicative of gastrointestinal cancer, including S100 calcium-binding protein P (), and revealed novel markers for GI cancer, including both and neuropeptide S receptor 1 (). Immunolocalization showed increased levels of LYZ, S100P, and NPSR1 proteins in UC and CAC. In conclusion, primary colonic organoid cultures from UC and non-IBD patients can be established that faithfully represent diseased or normal colonic states. These models reveal precancerous molecular pathways that are already activated in UC. The findings demonstrate the suitability of primary organoids for dissecting UC and CAC pathogenic mechanisms and suggest new targets for therapeutic intervention.
溃疡性结肠炎（UC）是炎症性肠病（IBD）的一种常见形式，其致病机制尚不清楚。阐明这些机制对于减轻UC症状以及预防UC进展为结肠炎相关结肠癌（CAC）至关重要。我们的目标是开发并验证可靠的、源自人类的UC模型，并在组织学、转录组学和表观遗传学水平上对其进行分析，以便对UC和CAC的发病机制进行机制研究。我们从UC和非IBD结肠上皮细胞中生成了患者来源的原代类器官培养物。我们对它们进行了组织学表型分析，并使用下一代测序方法对类器官和原代组织的全转录组和表观基因组进行了分析。组织结构以及粘蛋白2（MUC2）和溶菌酶（LYZ）的表达证明了类器官对健康和患病结肠上皮的组织学忠实性。转录组分析显示，UC患者来源的类器官和组织中炎症途径的表达增加。使用H3K27ac组蛋白修饰对活性增强子进行分析，发现UC来源的类器官富含指示胃肠道癌症的途径，包括S100钙结合蛋白P（），并揭示了胃肠道癌症的新标志物，包括和神经肽S受体1（）。免疫定位显示UC和CAC中LYZ、S100P和NPSR1蛋白水平升高。总之，可以建立来自UC和非IBD患者的原代结肠类器官培养物，它们能忠实地代表患病或正常的结肠状态。这些模型揭示了在UC中已经被激活的癌前分子途径。这些发现证明了原代类器官对于剖析UC和CAC致病机制的适用性，并为治疗干预提出了新的靶点。
