Project description
Studying R-loop formation in 2D and 3D tumor hypoxia models and its impact on cell survival
Hypoxia (reduced oxygen availability) is one of the hallmarks of solid tumors, caused by the difficulty of nutrient penetration through the deregulated tumor microenvironment. Hypoxic cells are highly resistant to standard chemotherapy and serve as a barrier to cancer treatment. Recent evidence shows that hypoxic cells have elevated levels of R-loops, three-stranded nucleic acid structures composed of an RNA:DNA hybrid and a displaced single-stranded DNA, formed when nascent RNA invades the DNA duplex. The main goal of the proposed project is to study the role and formation of R-loops in hypoxic cell biology and to explore whether R-loop processing factors could be used to enhance the effectiveness of cancer therapy. The project will be conducted simultaneously in a 2D cellular model within a hypoxic incubator and in a 3D cellular spheroid culture designed to mimic a hypoxic environment in solid tumors. It will utilize high-throughput microscopy to analyze hypoxia, DNA replication stress, DNA damage markers, and R-loop formation; mRNA sequencing to assess changes in transcriptional profiles; and ultimately, evaluate the sensitivity of hypoxic cells to inhibition of R-loop-processing factors.
