Title: Molecular mechanisms regulating cancer stem cell formation and maintenance in pancreatic cancer
Funder: Cancer Research UK

Abstract: •Background. Pancreatic ductal adenocarcinoma is one of the most lethal malignancies in human due to its highly metastatic characteristics and poor responsiveness to current cancer therapeutics. Pancreatic tumorigenesis involves a dedifferentiation process of cellular identity and the acquisition of a stem cell-like state of some cancer cells. These so-called cancer stem cells (CSCs) are exceptionally important because they can give rise to the whole tumour in the organism and lead to metastases. TGFß/Activin/Nodal-Smad2/3 developmental signalling is important for PDAC formation and mutations in this pathway are among the hallmarks of this cancer. Interestingly, TGFß/Activin/Nodal-Smad2/3 pathway controls the self-renewal of human pluripotent stem cells (hPSCs), and Nodal seems also to confer dedifferentiated stem cell-like features and metastatic potential to pancreatic CSCs by promoting expression of several pluripotency genes including Nanog. Therefore it is crucial to understand the mechanisms how the TGFß/Nodal-Smad2/3 signalling pathway governs pancreatic cancer stem cell formation and maintenance. • Aims. This programme will identify and characterize novel signal transduction / transcription pathways in pancreatic cancer stem cells. Its objectives are divided as follows. General objective: Characterize the function of TGFß/Nodal-Smad2/3 signalling in pancreatic CSCs. 1. Uncover Smad2/3 genome-wide targets and protein interactome in pancreatic CSCs and hPSCs. 2. Compare the function of Smad2/3 cofactors on self-renewal of CSCs and hPSCs. • Methods. My research will utilize state-of-the-art cell culture methods with genome-wide and proteomic techniques as well as in vivo PDAC formation experiments. I will apply a powerful combination of pancreatic CSC sphere cultures, PDAC organoids and hPSC-derived pancreatic progenitors for uncovering novel factors that cooperate with Smad2/3 in mediating stem cell-like cellular identity. I will identify the target loci of Smad2/3 by ChIP-sequencing and protein cofactors by SILAC / mass-spectrometry. Thereafter, I will characterize the role of selected Smad2/3 binding candidates on self-renewal of CSCs and hPSCs by loss-of-function and gain-of-function studies. Detailed mechanistic insight to candidate function will be obtained by promoter-luciferase assays, mapping of protein-protein interactions, and characterising their effects on epigenetic signatures and gene expression. Lastly, I will study their role in PDAC formation by orthotopic transplantation experiments in mice. • How the results of this research will be used. My research will uncover key mechanisms that trigger tissue transformation and cancer stem cell maintenance during PDAC formation. The arising discoveries can be imminently utilized for rational drug design by targeting CSC maintenance mechanisms for specifically eliminating these cells in cancer patients.


Research activity codes: 2.1 BIOLOGICAL AND ENDOGENOUS FACTORS;1.1 NORMAL BIOLOGICAL DEVELOPMENT AND FUNCTIONING
Health categories: CANCER AND NEOPLASMS