Circadian rhythms regulate diverse aspects of gastrointestinal physiology ranging from the composition of microbiota to motility. However, development of the intestinal circadian clock and detailed mechanisms regulating circadian physiology of the intestine remain largely unknown. In this report, we show that both pluripotent stem cell-derived human intestinal organoids engrafted into mice and patient-derived human intestinal enteroids possess circadian rhythms and demonstrate circadian phase-dependent necrotic cell death responses to Clostridium difficile toxin B (TcdB). Intriguingly, mouse and human enteroids demonstrate anti-phasic necrotic cell death responses to TcdB. RNA-Seq analysis shows that ~3–10% of the detectable transcripts are rhythmically expressed in mouse and human enteroids. Remarkably, we observe anti-phasic gene expression of Rac1, a small GTPase directly inactivated by TcdB, between mouse and human enteroids, and disruption of Rac1 abolishes clock-dependent necrotic cell death responses. Our findings uncover robust functions of circadian rhythms regulating clock-controlled genes in both mouse and human enteroids governing organism-specific, circadian phase-dependent necrotic cell death responses, and lay a foundation for human organ- and disease-specific investigation of clock functions using human organoids for translational applications.

中文翻译：

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肠道类器官生物钟的个体发育和功能


昼夜节律调节胃肠道生理学的各个方面，从微生物群的组成到蠕动。然而，肠道生物钟的发育和调节肠道昼夜节律生理学的详细机制仍然很大程度上未知。在这份报告中，我们表明，移植到小鼠体内的多能干细胞来源的人类肠道类器官和患者来源的人类肠道类器官都具有昼夜节律，并表现出对艰难梭菌毒素 B (TcdB) 的昼夜节律阶段依赖性坏死细胞死亡反应。有趣的是，小鼠和人类肠类物质表现出对 TcdB 的反阶段性坏死细胞死亡反应。 RNA-Seq 分析表明，约 3-10% 的可检测转录本在小鼠和人类肠类中有节律地表达。值得注意的是，我们观察到小鼠和人肠类之间Rac1 （一种被 TcdB 直接灭活的小型 GTP 酶）的反相基因表达，并且Rac1的破坏消除了时钟依赖性坏死细胞死亡反应。我们的研究结果揭示了小鼠和人类肠类中昼夜节律调节时钟控制基因的强大功能，这些基因控制着生物体特异性、昼夜节律阶段依赖性坏死细胞死亡反应，并为利用生物钟功能对人体器官和疾病特异性研究奠定了基础。用于转化应用的人类类器官。