Development and understanding of novel small molecules for cancer immunotherapies via REV-ERB regulation
File(s)
Author(s)
Uriz-Huarte, Amaia
Type
Thesis
Abstract
Immunotherapies for cancer have experienced an unprecedented breakthrough over the last decade, becoming one of the most powerful alternatives for cancer treatment. Natural Killer (NK) cells, members of the innate immune response, have risen as a promising strategy for the development of novel immunotherapies. They act upon diseased cells in a non-antigen dependant way and without the need to prior sensitization, with a short life-spam and less toxic cytokine secretion. All of this, potentially, translates into a faster, more potent and safer immunotherapeutic approach than some of the current trends in immunology.
REV-ERB is a nuclear receptor that has been identified as the direct regulator of E4bp4, the essential transcription factor involved in NK cells maturation. Inhibition of this regulator enhances the production of NK cells and thus represents a potential opportunity of improved cancer immunotherapy via NK cells.
REV-ERB’s role as part of the circadian rhythm has provided essential information regarding its mechanism of action and existing small molecule modulators. Our objective was to further the knowledge on REV-ERB modulation and develop a library of compounds that could allow us to control the expression of E4bp4 as a means to an NK cell-based immunotherapy.
As part of this thesis, two generations of REV-ERB ligands have been synthesised and several biological assays have been optimised for their characterisation, including reporter gene assay, viability screenings, ADME characterisation, CETSA, RT-qPCR, knockdown, EMSA and NK cell development analysis via flow cytometry. We have been able to further the understanding on REV-ERB ligands and we have successfully identified compounds with a better activity profile, improved stability, reduced toxicity, good selectivity for REV-ERB and with a positive effect at enhancing NK cell maturation.
Overall, this project has provided a strong foundation for the understanding of REV-ERB modulation and contributed towards the development of NK cell-based immunotherapies.
REV-ERB is a nuclear receptor that has been identified as the direct regulator of E4bp4, the essential transcription factor involved in NK cells maturation. Inhibition of this regulator enhances the production of NK cells and thus represents a potential opportunity of improved cancer immunotherapy via NK cells.
REV-ERB’s role as part of the circadian rhythm has provided essential information regarding its mechanism of action and existing small molecule modulators. Our objective was to further the knowledge on REV-ERB modulation and develop a library of compounds that could allow us to control the expression of E4bp4 as a means to an NK cell-based immunotherapy.
As part of this thesis, two generations of REV-ERB ligands have been synthesised and several biological assays have been optimised for their characterisation, including reporter gene assay, viability screenings, ADME characterisation, CETSA, RT-qPCR, knockdown, EMSA and NK cell development analysis via flow cytometry. We have been able to further the understanding on REV-ERB ligands and we have successfully identified compounds with a better activity profile, improved stability, reduced toxicity, good selectivity for REV-ERB and with a positive effect at enhancing NK cell maturation.
Overall, this project has provided a strong foundation for the understanding of REV-ERB modulation and contributed towards the development of NK cell-based immunotherapies.
Version
Open Access
Date Issued
2021-10
Date Awarded
2022-04
Copyright Statement
Creative Commons Attribution NonCommercial Licence
License URL
Advisor
Fuchter, Matthew
Publisher Department
Chemistry
Publisher Institution
Imperial College London
Qualification Level
Doctoral
Qualification Name
Doctor of Philosophy (PhD)