Division of General and Thoracic Surgery, The Hospital for Sick Children, University of Toronto, 1526-555 University Ave, Toronto, ON, M5G 1X8, Canada.
Division of Pediatric Surgery, Department of Surgery, Faculty of Medicine, Khon Kaen University, Khon Kaen, 40002, Thailand.
Pediatr Surg Int. 2020 Dec;36(12):1471-1479. doi: 10.1007/s00383-020-04765-2. Epub 2020 Oct 21.
The fibrogenic process in cholangiopathic diseases such as biliary atresia (BA) involves bile duct injury and apoptosis of cholangiocytes, which leads to the progression of liver fibrosis into liver cirrhosis and can result in end-staged liver disease. Recent advances in the development of organoids or mini-organ structures have allowed us to create an ex vivo injury model of the bile duct that mimics bile duct injury in BA. The aim of this experimental study was to develop a novel model of injured intrahepatic cholangiocytes as this can be relevant to BA. Our new model is important for studying the pathophysiological response of bile ducts to injury and the role of cholangiocytes in initiating the fibrogenic cascade. In addition, it has the potential to be used as a tool for developing new treatment strategies for BA.
Liver ductal organoids were generated from the liver of healthy neonatal mouse pups. Intrahepatic bile duct fragments were isolated and cultured in Matrigel dome. Injury was induced in the organoids by administration of acetaminophen in culture medium. The organoids were then evaluated for fibrogenic cytokines expression, cell apoptosis marker and cell proliferation marker.
Organoids generated from intrahepatic bile duct fragments organized themselves into single-layer epithelial spheroids with lumen on the inside mimicking in vivo bile ducts. After 24-h exposure to acetaminophen, cholangiocytes in the organoids responded to the injury by increasing expression of fibrogenic cytokines, transforming growth factor beta-1 (TGF-β1) and platelet-derived growth factor-BB (PDGF-BB). This fibrogenic response of injured organoids was associated with increased cholangiocyte apoptosis and decreased cholangiocyte proliferation.
To our knowledge this is the first description of cholangiocyte injury in the organoids derived from intrahepatic bile ducts. Our injury model demonstrated that cholangiocyte apoptosis and its fibrogenic response may play a role in initiation of the fibrogenic process in cholangiopathic diseases such as BA. These findings are important for the development of novel therapy to reduce cholangiocyte apoptosis and to halt the early fibrogenic cascade in liver fibrogenesis. This novel injury model can prove very valuable for future research in biliary atresia.
在胆病，如胆道闭锁（BA）等纤维发生过程中涉及胆管损伤和胆管细胞凋亡，这导致肝纤维化进展为肝硬化，并可能导致终末期肝病。最近在类器官或迷你器官结构的发展方面的进展使我们能够创建一种模拟 BA 中胆管损伤的胆管体外损伤模型。本实验研究的目的是开发一种新的损伤性肝内胆管细胞模型，因为这可能与 BA 有关。我们的新模型对于研究胆管对损伤的病理生理反应以及胆管细胞在启动纤维发生级联反应中的作用很重要。此外，它有可能被用作开发 BA 新治疗策略的工具。
从健康新生小鼠的肝脏中生成胆管类器官。分离肝内胆管片段并在 Matrigel 穹顶中培养。在培养物中用对乙酰氨基酚诱导类器官损伤。然后评估类器官中纤维生成细胞因子的表达、细胞凋亡标志物和细胞增殖标志物。
从肝内胆管片段生成的类器官自行组织成具有内部腔的单层上皮球体，模拟体内胆管。在 24 小时接触对乙酰氨基酚后，类器官中的胆管细胞通过增加纤维生成细胞因子，转化生长因子-β1（TGF-β1）和血小板衍生生长因子-BB（PDGF-BB）的表达来对损伤做出反应。这种损伤类器官的纤维发生反应与胆管细胞凋亡增加和胆管细胞增殖减少有关。
据我们所知，这是首次描述源自肝内胆管的类器官中的胆管细胞损伤。我们的损伤模型表明，胆管细胞凋亡及其纤维发生反应可能在胆病如 BA 中的纤维发生过程的启动中起作用。这些发现对于开发减少胆管细胞凋亡并阻止肝纤维化中早期纤维发生级联的新型治疗方法很重要。这种新的损伤模型对于未来的胆道闭锁研究将非常有价值。
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