A cholangiocyte organoid system for Cryptosporidium parvum cultivation and transcriptomic studies of biliary cryptosporidiosis  

作　　者：Miner Deng Qing Tian Xinjie Mao Jie Zhang Yanting Wei Na Li Yaqiong Guo Lihua Xiao Yaoyu Feng 

机构地区：[1]State Key Laboratory for Animal Disease Control and Prevention,South China Agricultural University,Guangdong,China [2]Guangdong Laboratory for Lingnan Modern Agriculture,Center for Emerging and Zoonotic Diseases,College of Veterinary Medicine,South China Agricultural University,Guangdong,China

出　　处：《hLife》2024年第7期360-370,共11页健康科学（英文）

基　　金：supported by National Natural Science Foundation of China(32030109);111 Project(D20008);Double First-class Discipline Promotion Project(2023B10564003);supported by National Natural Science Foundation of China(U21A20258);supported by Natural Science Foundation of Guangzhou City(202201010140)。

摘　　要：Biliary cryptosporidiosis caused by Cryptosporidium is life-threatening in immunocompromised individuals,particularly those with acquired immune deficiency syndrome(AIDS),resulting in AIDS cholangiopathy.However,studies of biliary cryptosporidiosis have lagged due to the lack of in vitro models that allow complete pathogen development.Here we describe procedures for the generation of mouse cholangiocyte organoids(Chol-orgs).The Chol-orgs,which expressed stem and biliary cell markers,could be clonally expanded for three months and stored in liquid nitrogen for more than one year.Combined with cell differentiation using the air-liquid interface(ALI)approach,we established a culture system for C.parvum.ALI cultures using Chol-orgs have supported50-fold amplification of the pathogen and generated viable oocysts in vitro.In addition,we analyzed the transcriptome of Chol-ALI cultures infected with the IId subtype of C.parvum to characterize host cell responses.RNA-seq analysis revealed that C.parvum upregulated immune and inflammatory responses and downregulated metabolic and cell proliferation signaling pathways in Cholorgs.A similar system using bovine cholangiocytes also supported the complete development of C.parvum in vitro.These in vitro models provide convenient methods to study biliary cryptosporidiosis and other hepatic infections and to develop effective therapies for AIDS cholangiopathy.

关 键 词：CRYPTOSPORIDIUM ORGANOIDS CHOLANGIOCYTE culture system host-pathogen interactions 

分 类 号：R318.1[医药卫生—生物医学工程]