RNA sequencing of pancreatic circulating tumour cells implicates WNT signalling in metastasis


Min Yu, David T. Ting, Shannon L. Stott, Ben S. Wittner, Fatih Ozsolak, Suchismita Paul, Jordan C. Ciciliano, Malgorzata E. Smas, Daniel Winokur, Anna J. Gilman, Matthew J. Ulman, Kristina Xega, Gianmarco Contino, Brinda Alagesan, Brian W. Brannigan, Patrice M. Milos, David P. Ryan, Lecia V. Sequist, Nabeel Bardeesy, Sridhar Ramaswamy, Mehmet Toner, Shyamala Maheswaran & Daniel A. Haber

 

Circulating tumour cells (CTCs) shed into blood from primary cancers include putative precursors that initiate distal metastases1. Although these cells are extraordinarily rare, they may identify cellular pathways contributing to the blood-borne dissemination of cancer. Here, we adapted a microfluidic device2 for efficient capture of CTCs from an endogenous mouse pancreatic cancer model3 and subjected CTCs to single-molecule RNA sequencing4, identifying Wnt2 as a candidate gene enriched in CTCs. Expression of WNT2 in pancreatic cancer cells suppresses anoikis, enhances anchorage-independent sphere formation, and increases metastatic propensity in vivo. This effect is correlated with fibronectin upregulation and suppressed by inhibition of MAP3K7 (also known as TAK1) kinase. In humans, formation of non-adherent tumour spheres by pancreatic cancer cells is associated with upregulation of multiple WNT genes, and pancreatic CTCs revealed enrichment for WNT signalling in 5 out of 11 cases. Thus, molecular analysis of CTCs may identify candidate therapeutic targets to prevent the distal spread of cancer