Biliary organoids uncover delayed epithelial development and barrier function in biliary atresia.
Biliary organoids uncover delayed epithelial development and barrier function in biliary atresia.
复制标题
胆道类器官揭示了胆道闭锁的上皮发育迟缓和屏障功能。
DOI：
10.1002/hep.32107
复制
发表时间：
2022-01
期刊：
影响因子：
--
通讯作者：
中科院分区：
文献类型：
--
作者：
胆道闭锁是一种病因不明的新生儿严重的炎症性和纤维化性胆管疾病。出生时出现的胆汁淤积症意味着产前出现了肝功能障碍。我们的目的是研究与异常胆管细胞发育有关的机制。我们从患有胆道闭锁的婴儿以及正常和患病对照的肝脏活检中生成胆道类器官。类器官从胆道闭锁的肝脏和对照组中出现，生长为含有管腔的球体，上皮内层是细胞角蛋白-19 -白蛋白阴性sox17 -阴性的胆管细胞样细胞。三组球的大体形态相似，表达了富含胆管细胞的基因。在胆道闭锁中，胆管细胞样细胞缺乏细胞核的基础定位，表达较少的发育和功能标记，并表现出纤毛的错误定向。他们异常表达F-actin， β-catenin和Ezrin，紧密连接蛋白zoonula occluden -1 （ZO-1）的信号较低，并且在维拉帕米治疗后，类器官内的罗丹明-123 （R123）信号较高，显示出通透性增加。胆道闭锁类器官中EGF信号和FGF2信号相关基因的表达降低。当用EGF+FGF2治疗时，胆道闭锁类器官表达分化（细胞角蛋白7和肝细胞核因子1同源盒B）和功能性（生长抑素受体2、囊性纤维化跨膜传导调节因子[CFTR]、水通道蛋白1）标记，恢复极性，改善F-actin、β-catenin和ZO-1的定位，增加CFTR功能，减少R123的摄取。胆道闭锁的类器官是有活力的，有证据表明上皮发育停止。EGF和FGF2诱导发育标志物，改善细胞间连接，降低上皮通透性，确定了促进上皮成熟和功能的潜在策略。
Biliary atresia is a severe inflammatory and fibrosing cholangiopathy of neonates of unknown etiology. The onset of cholestasis at birth implies a prenatal onset of liver dysfunction. Our aim was to investigate the mechanisms linked to abnormal cholangiocyte development. We generated biliary organoids from liver biopsies of infants with biliary atresia and normal and diseased controls. Organoids emerged from biliary atresia livers and controls and grew as lumen-containing spheres with an epithelial lining of cytokeratin-19posalbuminnegSOX17neg cholangiocyte-like cells. Spheres had similar gross morphology in all three groups and expressed cholangiocyte-enriched genes. In biliary atresia, cholangiocyte-like cells lacked a basal positioning of the nucleus, expressed fewer developmental and functional markers, and displayed misorientation of cilia. They aberrantly expressed F-actin, β-catenin, and Ezrin, had low signals for the tight junction protein zonula occludens-1 (ZO-1), and displayed increased permeability as evidenced by a higher Rhodamine-123 (R123) signal inside organoids after verapamil treatment. Biliary atresia organoids had decreased expression of genes related to EGF signaling and FGF2 signaling. When treated with EGF+FGF2, biliary atresia organoids expressed differentiation (cytokeratin 7 and hepatocyte nuclear factor 1 homeobox B) and functional (somatostatin receptor 2, cystic fibrosis transmembrane conductance regulator [CFTR], aquaporin 1) markers, restored polarity with improved localization of F-actin, β-catenin and ZO-1, increased CFTR function, and decreased uptake of R123. Organoids from biliary atresia are viable and have evidence of halted epithelial development. The induction of developmental markers, improved cell-cell junction, and decreased epithelial permeability by EGF and FGF2 identifies potential strategies to promote epithelial maturation and function.
登录
查看更多内容
影响因子：
17.1
作者：
Lorent K;Gong W;Koo KA;Waisbourd-Zinman O;Karjoo S;Zhao X;Sealy I;Kettleborough RN;Stemple DL;Windsor PA;Whittaker SJ;Porter JR;Wells RG;Pack M
通讯作者：
Pack M
影响因子：
14.8
作者：
Sampaziotis F;de Brito MC;Geti I;Bertero A;Hannan NR;Vallier L
通讯作者：
Vallier L
影响因子：
46.9
作者：
Ogawa, Mina;Ogawa, Shinichiro;Ghanekar, Anand
通讯作者：
Ghanekar, Anand
影响因子：
13.5
作者：
Omenetti, Alessia;Bass, Lee M.;Whitington, Peter F.
通讯作者：
Whitington, Peter F.
影响因子：
64.5
作者：
Huch M;Gehart H;van Boxtel R;Hamer K;Blokzijl F;Verstegen MM;Ellis E;van Wenum M;Fuchs SA;de Ligt J;van de Wetering M;Sasaki N;Boers SJ;Kemperman H;de Jonge J;Ijzermans JN;Nieuwenhuis EE;Hoekstra R;Strom S;Vries RR;van der Laan LJ;Cuppen E;Clevers H
通讯作者：
Clevers H