Investigating the synthetic lethal effects of poly (ADP-ribose) polymerase and histone deacetylase inhibition in progressive B-Cell chronic lymphocytic leukaemia deficient in the ATM pathway udand udOptimisation of multiple marker imaging of immunological tissue using confocal laser scanning fluorescence microscopy ud

摘要

This thesis contains two research projects. The first contributed to the development of poly (ADP-ribose) polymerase inhibitors (PARPis) and histone deacetylase inhibitors (HDACis) as targeted therapeutics for progressive, treatment-resistant forms of B-cell chronic lymphocytic leukaemia (B-CLL) exhibiting functional loss of the Ataxia-telangiectasia mutated (ATM) gene. ATM deficient cells exhibit genomic instability and defective homologous recombination (HR), an accurate DNA double-strand break repair mechanism. PARPis and HDACis impose cell dependency on HR by compromising other alternative DNA repair methods, resulting in lethal accumulation of DNA damage. The main hypothesis was that the two compounds combined would synergise and kill ATM deficient cells through this mechanism of synthetic lethality, however increased killing was not specifically observed in ATM-deficient CII and additionally TP53 mutant MEC-1 B-CLL cell lines compared to ATM wild type counterparts. The second project involved the development of a histopathological tissue imaging method, utilising indirect immunofluorescence and confocal laser-scanning microscopy (CLSM) to enable visualisation of multiple biomarkers within tissue cryosections. The simultaneous visualisation of the B-lymphocyte markers CD19, κ- and λ- immunoglobulin light chain antigen and a nuclear counterstain was achieved within reactive lymph node cryosections, and staining protocols were optimised for more photostable Alexa fluorophore - conjugated secondary antibodies.