Poster Presentation 38th Lorne Cancer Conference 2026

Pinpointing and targeting novel drivers of pancreatic cancer progression and metastasis using TRAP-seq (#266)

Michael Trpceski 1 2 , Alice Tran 1 , Nadia Kuepper 1 , Anna Howell 1 , Chi Kin (Kenny) Ip 1 2 , Cecilia Chambers 1 2 , Jef Servaes 1 , Shona Ritchie 1 2 , Jessie Zhu 1 2 , Daniel Reed 1 2 , Marjan Naeini 1 2 , Leonard Goldstein 1 2 , Grazi Vieira 1 , Robert Weatheritt 1 2 , Herbert Herzog 1 2 , Tatyana Chtanova 1 2 , Kendelle Murphy 1 2 , Paul Timpson 1 2 , David Herrmann 1 2
  1. The Garvan Institute of Medical Research, Sydney, New South Wales, Australia
  2. St Vincent’s Clinical School, University of New South Wales, Sydney, New South Wales, Australia

Pancreatic Cancer (PC) has a dismal 5-year survival rate of only 13%1, which can be attributed to its’ rapid metastatic spread and resistance to standard-of-care chemotherapy. Here, we aim to use innovative Translating-Ribosome-Affinity-Purification followed by RNA-sequencing (TRAP-seq) to assess the deregulated translatome of mutant p53-driven PC metastasis and identify potential novel therapeutic targets.

 

Our genetically engineered mouse models of PC are driven by an initiating KrasG12D mutation and either loss of p53 (p53flox mouse; poorly metastatic) or a gain-of-function mutation in p53 (p53R172H; KPC mouse; highly metastatic)2. They also express GFP-tagged ribosomes specifically in PC cells, which can then be immunoprecipitated with associated translating mRNAs for downstream RNA-seq.

 

As such, we isolated primary tumours from end-stage mice of both models, as well as matched KPC metastases. All cancer cells retain Rpl10a-GFP expression, and we confirmed the genomic presence of PC driver mutations. TRAP-seq of all samples resulted in a database representing a pool of over 1400 deregulated genes of the metastatic PC translatome. Four genes were subsequently identified as potential novel drivers of mutant p53-driven metastasis, with high expression of these genes being correlated with poor patient outcome. Validation confirmed increased expression of candidate genes in our discovery cohort as well as independent samples of highly metastatic PC when compared to poorly metastatic models. Pre-clinical in vitro functional assessments including proliferation, single-cell migration on cancer-associated fibroblast (CAF) derived matrices, cancer cell invasion in a CAF-contracted collagen matrix and anchorage-independent-growth assays have determined that genetic knock-down of candidate gene expression in KPC cancer cells significantly decreases cell migration, cell invasion and growth under anchorage-independent conditions, which can often be associated with a more aggressive cancer, while also significantly enhancing the response to standard-of-care gemcitabine/Abraxane chemotherapy. Recent in vivo intrasplenic transplant mouse models have also shown that candidate gene knockdown in KPC cancer cells result in a significant decrease in liver metastasis when compared to control. Further validation using our established in vivo PC orthotopic transplant mouse models is currently underway.

  1. Siegel, R.L., et al., Cancer statistics, 2025. CA: A Cancer Journal for Clinicians, 2025. 75(1): p. 10-45.
  2. Morton, J.P., et al., Mutant p53 drives metastasis and overcomes growth arrest/senescence in pancreatic cancer. Proceedings of the National Academy of Sciences, 2010. 107(1): p. 246-251.