Determining the efficacy and safety of cancer chemotherapeutics for cholangiocarcinoma (CCA) using Human Precision Cut Tissue Slices (hPCTS).

胆管癌（CCA）是一种胆管癌，是人类预后最差的恶性肿瘤之一。晚期诊断和有限的治疗选择意味着生存率很低，这是一种未满足需求的癌症。小鼠肝癌模型已经产生，以揭示驱动这种疾病的分子机制。基因突变可以通过化学或病毒诱导来促进肿瘤的形成，或者驱动恶性转化的关键途径可以被破坏或“敲除”，此外还有异种移植模型，即将肿瘤细胞植入免疫缺陷小鼠体内。然而，在体内的方法是昂贵的，耗时的，并引起伦理问题。因此，迫切需要研究胆管癌进展的体外替代方法。精确切割的肿瘤切片是可以体外培养的肝胆细胞的三维结构。它们概括了肝胆癌生物学的关键方面，并且至关重要的是保留了原始肿瘤微环境。在本提案中，我们建议在我们的实验室中实现使用来自人类肿瘤组织的精确切割组织切片（hPCTS）来研究驱动疾病进展的分子机制。我们的科学目标是在实验室中建立和验证肿瘤切片的生存能力，随着时间的推移维持肝脏和胆管的功能，并确保肿瘤突变在培养中保留下来。然后我们将评估hPCTS CCA对吉西他滨和顺铂的反应能力，这两种抗癌药物是胆管癌的标准治疗药物。我们将识别入侵的免疫细胞和肿瘤切片对癌症免疫疗法的反应能力。最后，我们将通过将hPCTS暴露于一组分子靶向抗癌药物来确定它们阻止肿瘤生长的能力，从而证实它们的临床相关性。总的来说，我们预计我们的方法将提供一个生物学相关的体外模型来了解胆管癌突变，这些突变可以用抗癌药物靶向，从而取代使用小鼠来研究这种疾病的重要生物学方面。

Cholangiocarcinoma (CCA) is a bile duct cancer and one of the worst outcome human malignancies. Late-stage diagnosis and limited therapeutic options means that survival rates are low and it is a cancer of unmet need.Mouse models of hepatobiliary cancer have been generated to unravel the molecular mechanisms that drive this disease. Genetic mutations can be induced chemically or virally to promote tumour formation, alternatively key pathways that drive malignant transformation can be disrupted or 'knocked out' , alongside xenograft models whereby tumour cells are implanted into immunodeficient mice. However, in vivo approaches are costly, time-consuming and raise ethical concerns. Therefore, in vitro alternatives to investigate bile duct cancer progression are urgently needed.Precision cut tumour slices are three-dimensional structures of hepatobiliary cells that can be cultured ex vivo. They recapitulate critical aspects of the hepatobiliary cancer biology and, crucially retain the original tumour microenvironment. In this proposal, we propose to implement the use of precision cut tissue slices derived from human tumour tissue (hPCTS) in our lab to study the molecular mechanisms driving disease progression. Our scientific goal is to establish and validate tumour slice viability in the lab and maintenance of liver and bile duct functionality over time and to ensure that the tumour mutations are retained in culture. We will then assess the ability of hPCTS CCA to respond to Gemcitabine and Cisplatin, anti-cancer drugs that are standard care for bile duct cancer. We will identify invading immune cells and the tumour slice's ability to respond to cancer immunotherapies. Finally, we will confirm their clinical relevance by exposing hPCTS to a panel of molecular targeted anti-cancer drugs to determine their ability to stop tumour growth.Overall, we anticipate that our approach will offer a biologically relevant in vitro model to understand bile duct cancer mutations which can be targeted with anti-cancer drugs, thereby replacing the use of mice to study vital biological aspects of this disease.