Murray, Benjamin Oliver; (2024) Development of an in vitro human bladder cancer model within a healthy urothelial context to use as a testbed for novel treatments. Doctoral thesis (Ph.D), UCL (University College London).

Text Murray_10198529_Thesis.pdf Access restricted to UCL open access staff until 1 November 2025. Download (18MB)

Abstract

Treatment outcomes for bladder cancer have not improved in the last 40 years. The gold-standard model for testing new therapies for bladder cancer is the PDX mouse model; however, there are physiological and molecular species differences in the bladder which may influence treatment responses. Therefore, more human-like models are sought to improve successful translation of promising new therapies into the clinic. In this thesis, I engineered an in vitro human bladder cancer model called the 3D-UHU-TU incorporating high- and low-grade cancer spheroids into the 3D-UHU healthy urothelial organoid model alongside a healthy urine environment similar to in vivo conditions. The 3D-UHU-TU models were characterised to ensure that they recapitulated low- and high-grade human bladder cancer. Cadherins, invasion markers, a proliferation marker and diagnostic markers were mostly expressed in the correct areas of the model and resembled that seen in human bladder cancer. One clinically known problem with Mitomycin C (MMC) treatment for bladder cancer is the lack of tissue penetration. Previously, Poly (lactic-co-glycolic) acid-encapsulated nitrofurantoin antibiotic microcapsules were proven to improve penetration and clear urinary tract infection in infected 3D-UHU models. Using this technology, with help from collaborators, I tested encapsulated MMC (known as ‘MitoCap’) on separated and full 3D-UHU-TU models. In both formulations tested, penetration of cargo was enhanced without impairing the therapeutic effect of MMC, but some normal cell cytotoxicity was observed. This result highlighted the need for more targeted therapies. Preliminary testing of alternative therapies including immunotherapy and oncolytic virus were performed on the model. Preliminary screening of bladder cancer-specific antigen NY-ESO-1, which would be useful for CAR-T cell therapy, was found to be expressed in the healthy 3D-UHU component as well as the tumour component, indicating there could be host damage with this treatment. PD-L1 screening was also tested for potential use with checkpoint inhibitors and also expressed in both components of the 3D-UHU-TU model. PD-L1 checkpoint inhibitors could still be tested but modifications to the model would be needed. Unfortunately, due to time constraints and logistical limitations, the immunotherapies could not be explored further, but may prove to be promising avenues in the future. I also tested a novel oncolytic virus (oHSV-GFP) designed by collaborators at University of Surrey on the 3D-UHU-TU models. oHSV-GFP successfully co-localised and replicated within cancer spheroids and not in healthy urothelium. The viruses also displayed specific cytotoxic effects in isolated cancer spheroids alone, and tumour-specific killing in 3D-UHU-TU models. Taken together, this thesis shows that the 3D-UHU-TU model can be used to test conventional and alternative therapies in a healthy urothelial environment. Models like this may one day lead to therapeutic innovations that could improve bladder cancer treatment outcomes.

Downloads since deposit

Loading...

3Downloads

Download activity - last month

Export as XMLCSVJSON

Loading...

Download activity - last 12 months

Export as CSVJSONXML

Loading...

Downloads by country - last 12 months

Export as XMLJSONCSV

Loading...

Archive Staff Only

View Item