| 研究実績の概要 |
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.
|
| 今後の研究の推進方策 |
We will analyze down-stream effects of TP53 and PBRM1 loss on iCOs in next experiments. We will also establish new organoids from human cholangiocarcinoma, develop a panel of immunohistochemistry makers to support the diagnosis of our histological-based TME classification of intrahepatic cholangiocarcinoma, evaluating clinical utility of the novel TME classification.
|
