Advancing Phenotypic Drug Discovery for Cardiac Hypertrophy : Development and Application of a High-Throughput and High-Content Screening Platform
This thesis presents a high-throughput phenotypic screening platform tailored for cardiac hypertrophy that enables efficient lead optimisation and facilitates translation into in vivo models. A mixed neonatal rat cardiac cell culture was established to analyse cardiomyocytes and non-cardiomyocytes responses in parallel. A self-developed CellProfiler pipeline generated a multi-parametric 'Hypertrophy Score' for phenotypic evaluation. The platform was first applied to investigate a G protein-coupled receptor kinase 5 (GRK5) inhibitor library. The assay platform generated meaningful cellular structure-activity relationships (SAR) to correlate with biochemical IC50 values and physicochemical features. GRK5 inhibitor 4a emerged as a highly effective, non-toxic anti-hypertrophic agent, with its inactive optical antipode 4b serving as an ideal negative control. Secondly, the role of L-2-hydroxyglutarate (L-2HG) as a novel pro-hypertrophic metabolite was investigated. Given its metabolic link to glutamine, small-molecule inhibitors targeting glutaminase (GLS1) and glutamine uptake were systematically profiled for their anti-hypertrophic and anti-proliferative effects. Lastly, the screening platform was leveraged to explore the tumour-heart axis by assessing cardiomyocyte responses to serum from colorectal cancer patients. Patient serum induced proliferative effects across cell populations but suppressed hypertrophic markers, suggesting circulating tumour-derived factors with cardiodepressive properties. Collectively, this thesis introduces a novel multiparametric platform for cardiac drug discovery, demonstrating the power of combining high-content phenotyping, translational models and targeted interventions to address unmet clinical needs in cardiac hypertrophy.
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