E-Poster Presentation 33rd Lorne Cancer Conference 2021

Generation and validation of C57BL/6 syngeneic pancreatic cancer-associated fibroblasts as a new tool to study stromal and immune interactions in pancreatic cancer (#157)

Cecilia Chambers 1 , Michael Trpceski 1 , Shona Ritchie 1 , Daniel Reed 1 , Victoria Lee 1 , Lea Abdulkhalek 1 , Kendelle Murphy 1 , Max Nobis 1 , Brooke Pereira 1 , Paul Timpson 1 , David Hermann 1
  1. The Garvan Institute of Medical Research, Sydney, NSW, Australia

Due to surgically unresectable, locally advanced or metastatic disease being present at the time of clinical diagnosis, pancreatic cancer (PC) is one of the most lethal forms of human cancer, with >90% of patient deaths occurring within 1 year of diagnosis1. Consequently, the development of more effective strategies to overcome these limitations and efficiently treat PC is required.

 

Extensive fibrosis is one of the cornerstone hallmarks of PC tumorigenesis, metastasis and acquired therapy resistance2. This severe fibrotic response is characterised by the recruitment and subsequent activation of cancer-associated fibroblasts (CAFs), abnormal extracellular matrix (ECM) assembly and remodelling, increased inflammatory response, distorted or compromised immune responses and altered angiogenesis and blood supply. Upon recruitment by cancer cells, CAFs produce factors that can alter ECM structure, support cancer cell motility and survival and suppress immune surveillance2,3. The genetically engineered KPC mouse model of PC (Pdx1-Cre; KrasG12D/+; p53R172H/+) closely mimics the mutational landscape, histopathology and progression of the human disease as well as the associated stromal and immunological dysfunction, making it an excellent model to study PC in a preclinical context4.  

 

We have backcrossed the KPC mouse model to the C57BL/6 genetic background for 10 generations and isolated syngeneic cancer cells and CAFs from end-stage pancreatic tumours. Using known cancer cell and CAF markers, we confirmed the identity of both cell lines via immunofluorescence imaging and quantitative RT-qPCR. Importantly, our syngeneic cancer cells and CAFs are immunologically compatible with immunocompetent C57BL/6 mice and in a subcutaneous co-injection model establish tumours, which may more accurately reflect the dense desmoplastic nature/features of the disease. Our syngeneic CAFs in conjunction with syngeneic PC cells will therefore serve as an exciting novel tool from which to assess the complex interactions between PC cells, stroma and the immune system. Investigating these interactions may provide new targets for therapy that could boost immune surveillance and immune cell-mediated killing of PC cells.

  1. 1. Siegel, R.L et al. (2020) Cancer statistics, 2020. CA Cancer J. Cin. 70 (1), 7-30.
  2. 2. Pereira, B.A et al. (2019) CAF subpopulations: a new reservoir of stromal targets in pancreatic cancer. Trends in Cancer. 5 (11), 724-741.
  3. 3. Elyada, E et al. (2019) Cross-species single-cell analysis of pancreatic ductal adenocarcinoma reveals antigen-presenting cancer-associated fibroblasts. Cancer Discovery. 9 (8), 1102-1123.
  4. 4. Morton, J.P et al. (2010) Mutant p53 drives metastasis and overcomes growth arrest/senescence in pancreatic cancer. PNAS. 107 (1), 246-251.