Sen Akira, Imai Atsuki, Miyauchi Eiji, Yanagisawa Kota, Oda Tsukasa, Sasaki Fuki, Uchida Shintaro, Okada Takuhisa, Yokobori Takehiko, Saeki Hiroshi, Odamaki Toshitaka, Sasaki Nobuo
The Laboratory for Mucosal Ecosystem Design, Institute for Molecular and Cellular Regulation, Gunma University, Maebashi, 371-8512, Gunma, Japan.
Innovative Research Institute, Morinaga Milk Industry Co Ltd, Zama, 252-8583, Kanagawa, Japan.
Sci Rep. 2025 Jul 1;15(1):22325. doi: 10.1038/s41598-025-07391-x.
The large intestine has a dense milieu of indigenous bacteria, generating a complex ecosystem with crosstalk between individual bacteria and host cells. In vitro host cell modeling and bacterial interactions at the anaerobic interphase have elucidated the crosstalk molecular basis. Although classical cell lines derived from patients with colorectal cancer including Caco-2 are used, whether they adequately mimic normal colonic epithelial physiology is unclear. To address this, we performed transcriptome profiling of Caco-2 and Monolayer-cultured epithelial cells derived from healthy Human Colonic Organoids (MHCO) cultured hemi-anaerobically. Coculture with the anaerobic gut bacteria, Bifidobacterium longum subsp. longum differentiated the probiotic effects of test cells from those of physiologically normal intestinal and colorectal cancer cells. We cataloged non- or overlapping gene signatures where gene profiles of Caco-2 represented absorptive cells in the small intestinal epithelium, and MHCO showed complete colonic epithelium signature, including stem/progenitor, goblet, and enteroendocrine cells colonocytes. Characteristic gene expression changes related to lipid metabolism, inflammation, and cell-cell adhesion were observed in cocultured live Bifidobacterium longum and Caco-2 or MHCO. B. longum-stimulated MHCO exhibited barrier-enhancing characteristics, as demonstrated in clinical trials. Our data represent a valuable resource for understanding gut microbe and host cell communication.
大肠拥有密集的本土细菌环境，形成了一个复杂的生态系统，其中单个细菌与宿主细胞之间存在相互作用。体外宿主细胞建模以及在厌氧界面处的细菌相互作用已经阐明了这种相互作用的分子基础。尽管使用了源自包括Caco-2在内的结直肠癌患者的经典细胞系，但它们是否能充分模拟正常结肠上皮生理仍不清楚。为了解决这个问题，我们对来自半厌氧培养的健康人结肠类器官（MHCO）的Caco-2和单层培养上皮细胞进行了转录组分析。与厌氧肠道细菌长双歧杆菌亚种共培养，区分了测试细胞与生理正常的肠道和结直肠癌细胞的益生菌作用。我们编目了非重叠或重叠的基因特征，其中Caco-2的基因谱代表小肠上皮中的吸收细胞，而MHCO显示出完整的结肠上皮特征，包括干细胞/祖细胞、杯状细胞和肠内分泌细胞结肠细胞。在共培养的活长双歧杆菌与Caco-2或MHCO中观察到了与脂质代谢、炎症和细胞间粘附相关的特征性基因表达变化。如临床试验所示，长双歧杆菌刺激的MHCO表现出增强屏障的特性。我们的数据为理解肠道微生物与宿主细胞通讯提供了宝贵资源。
