Pathogenesis roles and therapeutic targets of PBRM1 mutation in intrahepatic cholangiocarcinoma

我们从非肿瘤肝脏中创建了12个肝内胆管细胞类器官（iCO）系，并优化了CHO和肝内胆管癌类器官（iCCAO）的条件。正常ico在ROCK抑制剂（Y-27632）和R-Spondin的培养条件下生长良好，不含Wnt3a。我们建立了03个PBRM1敲除（KO）的iCCA细胞系，在正常的iCO上逐步进行TP53和PBRM1的KO，并利用CRISPR/Cas9建立了TP53-KO iCO，以及双TP53和PBRM1-KO iCO。一种新的iCCA TME分类已经建立，包括四种亚型，表现出不同的T细胞、骨髓细胞、CAFs、细胞外基质（如胶原）和空间蛋白质组学分析的基因表达模式，包括CTLA4、PD-1、VISTA、LAG3和TAGLN。

We have created 12 lines of intrahepatic cholangiocyte organoids (iCO)from non-tumor livers and optimized conditions for CHO and intrahepatic cholangiocarcinoma organoids (iCCAO). Normal iCOs were grown well in our culture condition with ROCK Inhibitor (Y-27632) and R-Spondin, without Wnt3a. We have created 03 iCCA cell lines with PBRM1 knockout (KO), performed step-by-step TP53 and PBRM1 KO on normal iCOs, and established TP53-KO iCO, and double TP53 and PBRM1-KO iCOs, using CRISPR/Cas9.A novel TME classification of iCCA has been established, including four subtypes, showing distinct organizing patterns of T cells, myeloid cells, CAFs, an extracellular matrix such as collagen, and gene expression on spatial proteomic profiling, including CTLA4, PD-1, VISTA, LAG3, and TAGLN.