A new bladder cancer model based on tissue reprogramming and gene targeting


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Flaminia Talos and Daifeng Wang

With recent advances in cellular reprogramming and gene editing it became pos-sible to envision new approaches for tissue modeling in normal and disease contexts. Specifically, we propose to use transdifferentiation and gene targeting to generate a novel genetically-engineered model system for studies of human cancer. We recently devel-oped a highly innovative methodology for generating fully functional prostate tissue in renal grafts based on a computational system approach that identifies synergistic speci-fication genes (Talos et al., Nature Commun, 2017). We propose here to apply and ex-pand these methods for modeling bladder cancer by combining lineage conversion of fibroblasts with tissue recombination assays, advanced computational systems biology algorithms and CRIPSR/Cas9-mediated gene targeting of clinically-relevant mutations. In our preliminary studies, we have shown that fibroblasts can be directly converted into epithelial cells following transient expression of the pluripotency factors in pro-epithelial culture conditions. Moreover, these induced epithelial cells are amenable to further ter-minal differentiation into bladder tissue in tissue recombination assays in vivo under the inductive force of bladder specific mesenchyme. We have also employed computational algorithms to infer master regulators of urothelium development. Based on these prelim-inary data, we hypothesize that the inherent plasticity of readily-accessible fibroblasts can be exploited to generate bladder epithelia through a combination of key bladder specifi-cation genes, reprogramming techniques and tissue recombination assays. Moreover, we hypothesize that the reprogrammed bladder tissue is amenable to malignant transfor-mation through CRISPR-mediated gene targeting.  

To test this hypothesis and generate a new model of human cancer, we propose to (1) Convert human fibroblasts to bladder epithelium by activating the developmental regulatory genes and (2) Model bladder cancer by CRISPR-mediated gene targeting in the reprogrammed tissue of tumor suppressors and oncogenes relevant for human dis-ease. Our studies will provide novel insights into the mechanisms underlying bladder tu-morigenesis and a novel platform for drug screening and for discovery of patient-specific early prognostic biomarkers.  

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