Introduction/Background The prognosis for patients with platinum-resistant ovarian High Grade Serous Carcinoma (HGSC) remains poor. Data from the BriTROC-1 study indicate that genomic alterations alone cannot explain acquired platinum resistance in many cases, and emerging evidence suggests epigenetic alterations may be critical. We wish to investigate epigenetic changes that may drive platinum resistance in HGSC by treating established HGSC cell lines and patient-derived cells with pulses of carboplatin and investigating the nature, kinetics and plasticity of platinum-induced epigenetic changes.
Methodology We will mimic, using in an in vitro two-dimensional model, multiple cycles of platinum-based chemotherapy as used clinically. We will generate preliminary results from established cell lines and primary cultures. The primary cell cultures are collected from the ascites of patients with HGSC treated at Imperial College NHS Trust, London. Following validation (p53, PAX8 immunocytochemistry), carboplatin sensitivity is assessed (sulforhodamine B assay). Cells are then pulsed with four cycles of carboplatin (50μM for 6 hours) with a week of recovery between each cycle. Chemosensitivity of surviving cells is measured after each cycle. The cells are then harvested for downstream methylation (Illumina 850k array), transcriptomic (RNA sequencing) and chromatin accessibility (ATAC sequencing) assays. Cells are also imaged using STORM (Stochastic Optical Reconstruction Microscopy). Preliminary STORM data already indicate differences in chromatin structure and the distribution of specific histone modifications between paired sensitive and resistant HGSC cell lines.
Results We will receive the raw data within 8–12 weeks from now for the bioinformatic analysis. Differential gene expression analysis will uncover differently enriched pathways under the selective pressure of platinum-based chemotherapy.
Conclusion Understanding the epigenetic landscape of HGSC in real time using physiologically relevant models will allow us to identify possible therapeutic targets that could eventually prevent platinum resistance.
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