Modelling Candida albicans infection of the human gut using human intestinal organoid cultures

白色念珠菌是公认的人类最重要的真菌病原体。C.白色念珠菌完全适应于健康哺乳动物宿主的寄生虫病，但当宿主出现免疫缺陷、微生物生态失调和上皮损伤时发生发病。人肠上皮细胞对C.对白色念珠菌感染的认识还不十分清楚。肠腔因子，如释放到管腔微环境中的饮食来源和微生物代谢产物，可能是易感性的驱动因素。目前，研究不得不依赖于小鼠模型或人类上皮细胞系;然而，这些并不完全模拟人类肠道。在该项目中，将使用人源性肠道类器官，取代小鼠模型的使用。肠道对真菌感染易感性的一个潜在因素是一种强效的肠道毒素，即常见的真菌毒素食物污染物脱氧雪腐镰刀菌烯醇（DON）。DON靶向人上皮细胞系和猪肠外植体中的肠上皮屏障完整性和促炎信号传导。此外，DON暴露已被证明促进肠道细菌移位和病原菌如沙门氏菌和大肠杆菌的全身感染。coli的体外和体内培养。本项目旨在进一步发展我们的人源性肠道类器官模型，以研究C。白念珠菌与人肠道的接触，阐明真菌毒素DON对人肠道上皮细胞对白念珠菌易感性的影响。白色念珠菌感染目的：1.建立肠道类器官模型，研究C.白色念珠菌与人肠上皮细胞的关系。研究了C.白念珠菌对肠屏障功能和促炎反应的影响.研究饮食来源的肠道真菌毒素（脱氧雪腐镰刀菌烯醇，DON）对上皮细胞对C。方法学：该项目将利用人源性肠类器官培养物来评估肠粘连和念珠菌感染。白念珠菌，并探讨DON的潜在影响。将建立宿主-真菌共培养物，并使用跨上皮电阻、屏障蛋白表达和细胞因子释放的诱导来评估肠功能。

Candida albicans is recognised as the most prominent fungal commensal and pathogen of humans. C. albicans is perfectly adapted for commensalism in healthy mammalian hosts, but pathogenesis occurs when hosts develop immunodeficiency, microbial dysbiosis and epithelial damage. Susceptibility of the human intestinal epithelium to C. albicans infection is not well understood. It is likely that intestinal luminal factors, such as diet-derived and microbial metabolites released into the luminal microenvironment, act as drivers of susceptibility. At present, studies have had to rely upon mouse models or human epithelial cell lines; however, these do not completely mimic the human gut. In this project human-derived intestinal organoids will be utilised, REPLACING use of mouse models. One potential contributor to intestinal susceptibility to fungal infection is a potent intestinal toxin, the common mycotoxin food contaminant deoxynivalenol (DON). DON targets intestinal epithelial barrier integrity and pro-inflammatory signalling in human epithelial cell lines and pig intestinal explants. Furthermore, DON exposure has been shown to facilitate intestinal bacterial translocation and systemic infection by pathogenic bacteria such as Salmonella and E. coli in vitro and in vivo. This project aims to further develop our human-derived intestinal organoid model to investigate interactions of C. albicans with the human gut and to elucidate the effect of the mycotoxin DON on the susceptibility of the human intestinal epithelium to C. albicans infection. Aims:1. Develop intestinal organoids model to study the interaction of C. albicans with human gut epithelium.2. Study the effect of C. albicans on intestinal barrier function and pro-inflammatory responses.3. Investigate the effect of a diet-derived intestinal mycotoxin (deoxynivalenol, DON) on epithelial responses to C. albicans.Methodology:The project will utilise human-derived intestinal organoid cultures to assess the intestinal adhesion and infection of C. albicans and to explore the potential effects of DON. Host-fungal co-cultures will be established and intestinal function will be assessed as using trans epithelial electrical resistance, barrier protein expression and induction of cytokine release.