Department of Pediatrics, University of California, San Diego, La Jolla, California.
Department of Medicine, University of California, San Diego, La Jolla, California.
Am J Physiol Gastrointest Liver Physiol. 2019 Nov 1;317(5):G580-G591. doi: 10.1152/ajpgi.00098.2019. Epub 2019 Aug 21.
Congenital tufting enteropathy (CTE) is an autosomal recessive disease characterized by severe intestinal failure in infancy and mutations in the epithelial cell adhesion molecule () gene. Previous studies of CTE in mice expressing mutant EpCAM show neonatal lethality. Hence, to study the cellular, molecular, and physiological alterations that result from EpCAM mutation, a tamoxifen-inducible mutant EpCAM enteroid model has been generated. The presence of mutant EpCAM in the model was confirmed at both mRNA and protein levels. Immunofluorescence microscopy demonstrated the reduced expression of mutant EpCAM. Mutant enteroids had reduced budding potential as well as significantly decreased mRNA expression for epithelial lineage markers (), proliferation marker and secretory pathway transcription factors (, . Significantly decreased numbers of Paneth and goblet cells were confirmed by staining. These findings were correlated with intestinal tissue from CTE patients and the mutant mice model that had significantly fewer Paneth and goblet cells than in healthy counterparts. FITC-dextran studies demonstrated significantly impaired barrier function in monolayers derived from mutant enteroids compared with control monolayers. In conclusion, we have established an ex vivo CTE model. The role of EpCAM in the budding potential, differentiation, and barrier function of enteroids is noted. Our study establishes new facets of EpCAM biology that will aid in understanding the pathophysiology of CTE and role of EpCAM in health and disease. Here, we develop a novel ex vivo enteroid model for congenital tufting enteropathy (CTE) based on epithelial cell adhesion molecule ( gene mutations found in patients. With this model we demonstrate the role of EpCAM in maintaining the functional homeostasis of the intestinal epithelium, including differentiation, proliferation, and barrier integrity. This study further establishes a new direction in EpCAM biology that will help in understanding the detailed pathophysiology of CTE and role of EpCAM.
先天性簇状肠病（CTE）是一种常染色体隐性疾病，其特征是婴儿期严重的肠道衰竭和上皮细胞黏附分子（）基因突变。以前在表达突变 EpCAM 的小鼠中对 CTE 的研究表明，新生儿死亡率很高。因此，为了研究 EpCAM 突变导致的细胞、分子和生理改变，生成了一个可诱导突变 EpCAM 肠类器官模型。在该模型中，在 mRNA 和蛋白质水平上都证实了突变 EpCAM 的存在。免疫荧光显微镜显示突变 EpCAM 的表达减少。突变肠类器官的出芽潜力降低，上皮谱系标志物（）、增殖标志物和分泌途径转录因子（、的 mRNA 表达显著降低。通过染色证实 Paneth 和杯状细胞的数量显著减少。这些发现与 CTE 患者的肠道组织和突变小鼠模型相关联，与健康对照相比，突变小鼠模型中的 Paneth 和杯状细胞数量明显减少。FITC-葡聚糖研究表明，与对照单层相比，突变肠类器官衍生的单层中屏障功能显著受损。总之，我们建立了一个体外 CTE 模型。注意到 EpCAM 在肠类器官出芽潜力、分化和屏障功能中的作用。我们的研究确定了 EpCAM 生物学的新方面，将有助于理解 CTE 的病理生理学以及 EpCAM 在健康和疾病中的作用。在这里，我们基于在患者中发现的上皮细胞黏附分子（基因突变，开发了一种用于先天性簇状肠病（CTE）的新型体外肠类器官模型。通过该模型，我们证明了 EpCAM 在维持肠道上皮的功能平衡中的作用，包括分化、增殖和屏障完整性。这项研究进一步确定了 EpCAM 生物学的一个新方向，将有助于理解 CTE 的详细病理生理学以及 EpCAM 的作用。
