Determining the efficacy and safety of cancer chemotherapeutics for cholangiocarcinoma (CCA) using Human Precision Cut Tissue Slices (hPCTS).
Lead Research Organisation:
Liverpool John Moores University
Department Name: Pharmacy and Biomolecular Sciences
Abstract
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.
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.
Technical Summary
Despite advances in our understanding of the molecular pathogenesis of cholangiocarcinoma (CCA), 5-year survival rates have remained unchanged in recent decades (~5%). There is an urgent, unmet need for understanding the molecular basis of this disease and to develop novel targeted therapies.
Mouse models of hepatobiliary cancer have enabled a comprehensive investigation of the genetics and biology of this disease. However, in vivo approaches are costly and time-consuming, and crucially raise ethical
concerns. Precision cut tissue slices (PCTS) offer an alternative as they retain all cell types in their natural 3D tissue environment, in the correct proportions and orientations. Recent modifications to media composition and incubation conditions have improved slice viability and maintenance of phenotype over time.
The aim of this proposal is to establish and validate the use of the use of human PCTS from resected CCA tumour tissue, as an in vitro model to study the molecular pathogenesis of CCA, and assess their relevance to perform therapeutic studies. This technique will replace the need to conduct animal experiments to investigate disease progression and provide a translational model, highlighted in the 2020 CCA consensus statement as an urgent requirement.
Our scientific goal is to validate tumour slice viability, maintenance of hepatobiliary functionality and tumour mutational burden ex vivo. We will then assess the relevance of hPCTS CCA to perform therapeutic studies utilising Gemcitabine/Cisplatin (first line therapy) alongside involvement of invading immune cells and the tumour microenvironment to assess their ability to respond to immunotherapies. Finally, we will confirm their clinical relevance by exposing hPCTS to a panel of chemotherapeutics to determine their efficacy and safety alongside targeted therapies directed by CCA mutational analysis recently identified in our applicant group to inform clinical strategy and management.
Mouse models of hepatobiliary cancer have enabled a comprehensive investigation of the genetics and biology of this disease. However, in vivo approaches are costly and time-consuming, and crucially raise ethical
concerns. Precision cut tissue slices (PCTS) offer an alternative as they retain all cell types in their natural 3D tissue environment, in the correct proportions and orientations. Recent modifications to media composition and incubation conditions have improved slice viability and maintenance of phenotype over time.
The aim of this proposal is to establish and validate the use of the use of human PCTS from resected CCA tumour tissue, as an in vitro model to study the molecular pathogenesis of CCA, and assess their relevance to perform therapeutic studies. This technique will replace the need to conduct animal experiments to investigate disease progression and provide a translational model, highlighted in the 2020 CCA consensus statement as an urgent requirement.
Our scientific goal is to validate tumour slice viability, maintenance of hepatobiliary functionality and tumour mutational burden ex vivo. We will then assess the relevance of hPCTS CCA to perform therapeutic studies utilising Gemcitabine/Cisplatin (first line therapy) alongside involvement of invading immune cells and the tumour microenvironment to assess their ability to respond to immunotherapies. Finally, we will confirm their clinical relevance by exposing hPCTS to a panel of chemotherapeutics to determine their efficacy and safety alongside targeted therapies directed by CCA mutational analysis recently identified in our applicant group to inform clinical strategy and management.
