Mapping tumour heterogeneity and predictive cancer signatures in vivo
Purpose: In order to follow the fate of the bladder cells, we will use a mouse model expressing the Cre recombinase under the control of the uroplakin 2 (Upk2), a bladder specific promoter actively expressed in the bladder urothelium (Tg(Upk2-Cre)1Rkl/WghJ Jax Stock No.029281). By crossing this line with R26-Confetti (Jax Stock No.013731), the urothelial cells will be specifically and irreversibly labelled with a fluorescent marker. Once fluorescently labelled, a specific cell type will permanently retain this mark, even if it undergoes proliferation or a drastic change in cell fate (such as trans-differentiation or de-differentiation), thus enabling cell fate tracing, cell sorting and further cell characterisation.
Expected distress for the animals: weight loss, hematuria. Male mice develop more tumours than female mice, therefore groups should be analysed separately.
Expected scientific or societal benefit: Our project aim at characterising the unknown mechanisms governing the bladder urothelial disease. We expect our research to: (1) strongly contribute to the general data base of knowledge by depositing all the omics data in public repositories following publication thus allowing further analysis and integration from other groups addressing other biological questions; (2) decisively refine the biological interpretations, by generating the spatial and temporal dynamic map of cell identity maintenance pathways dynamic in tumours and apparently normal tissue (3) uncover new strategies for tumour control by pinpointing a series of cell-plasticity targets involved in tumour initiation and progression, that can be further validated through preclinical and clinical research, hence bearing potential clinical impact in cancer treatment. (4) potentially find a common cell-plasticity switch controlling the tumour formation between different cancers. This will have an obvious clinical and social impact with repercussions on the current targeted therapies.
Number of animals to be used: a total of 800 UPK2-cre; R26-Confetti transgenic mice.
Replacement, reduction and improvement
All experiments that do not require a living organism (lineage tracing; metabolism; systemic organ interactions) will be performed in vitro by using patient-derived hiPSC or on isolated human tumour cells. We will optimise the number of mice necessary for this project by using common experimental groups, where the same set of collected samples will be processed separately for replying distinct questions, hence halving the number of required animals.
Expected distress for the animals: weight loss, hematuria. Male mice develop more tumours than female mice, therefore groups should be analysed separately.
Expected scientific or societal benefit: Our project aim at characterising the unknown mechanisms governing the bladder urothelial disease. We expect our research to: (1) strongly contribute to the general data base of knowledge by depositing all the omics data in public repositories following publication thus allowing further analysis and integration from other groups addressing other biological questions; (2) decisively refine the biological interpretations, by generating the spatial and temporal dynamic map of cell identity maintenance pathways dynamic in tumours and apparently normal tissue (3) uncover new strategies for tumour control by pinpointing a series of cell-plasticity targets involved in tumour initiation and progression, that can be further validated through preclinical and clinical research, hence bearing potential clinical impact in cancer treatment. (4) potentially find a common cell-plasticity switch controlling the tumour formation between different cancers. This will have an obvious clinical and social impact with repercussions on the current targeted therapies.
Number of animals to be used: a total of 800 UPK2-cre; R26-Confetti transgenic mice.
Replacement, reduction and improvement
All experiments that do not require a living organism (lineage tracing; metabolism; systemic organ interactions) will be performed in vitro by using patient-derived hiPSC or on isolated human tumour cells. We will optimise the number of mice necessary for this project by using common experimental groups, where the same set of collected samples will be processed separately for replying distinct questions, hence halving the number of required animals.