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Updated: Oct 28, 2025
Genome-Wide CRISPR Screen for Unveiling Radiosensitive and Radioresistant Genes
Published on: May 23, 2025
Three-dimensional CRISPR screening reveals epigenetic interaction with anti-angiogenic therapy
Michael Y He1,2,3, Michael M Halford1, Ruofei Liu1,2
1Tumour Angiogenesis and Microenvironment Program, Peter MacCallum Cancer Centre, Melbourne, VIC, Australia.
Abstract:
Angiogenesis underlies development, physiology and pathogenesis of cancer, eye and cardiovascular diseases. Inhibiting aberrant angiogenesis using anti-angiogenic therapy (AAT) has been successful in the clinical treatment of cancer and eye diseases. However, resistance to AAT inevitably occurs and its molecular basis remains poorly understood. Here, we uncover molecular modifiers of the blood endothelial cell (EC) response to a widely used AAT bevacizumab by performing a pooled genetic screen using three-dimensional microcarrier-based cell culture and CRISPR-Cas9. Functional inhibition of the epigenetic reader BET family of proteins BRD2/3/4 shows unexpected mitigating effects on EC survival and/or proliferation upon VEGFA blockade. Moreover, transcriptomic and pathway analyses reveal an interaction between epigenetic regulation and anti-angiogenesis, which may affect chromosomal structure and activity in ECs via the cell cycle regulator CDC25B phosphatase. Collectively, our findings provide insight into epigenetic regulation of the EC response to VEGFA blockade and may facilitate development of quality biomarkers and strategies for overcoming resistance to AAT.
Insights
Researchers identified epigenetic regulators, specifically BET proteins, that influence endothelial cell survival during anti-angiogenic therapy (AAT). Understanding this epigenetic regulation may help overcome resistance to AAT in cancer and eye diseases.
Area of Science:
- Cell Biology
- Molecular Biology
- Epigenetics
Background:
- Angiogenesis is crucial for development and disease, including cancer, eye, and cardiovascular conditions.
- Anti-angiogenic therapy (AAT) is clinically successful but faces inevitable resistance.
- The molecular mechanisms underlying AAT resistance are not fully understood.
Purpose of the Study:
- To identify molecular modifiers of blood endothelial cell (EC) response to anti-angiogenic therapy (AAT).
- To investigate the role of epigenetic regulation in AAT resistance.
Main Methods:
- Pooled genetic screening using CRISPR-Cas9 technology.
- Three-dimensional microcarrier-based cell culture.
- Transcriptomic and pathway analyses.
Main Results:
- Functional inhibition of BET proteins (BRD2/3/4) mitigated EC survival and proliferation under VEGFA blockade.
- Epigenetic regulation interacts with anti-angiogenesis pathways.
- Potential involvement of cell cycle regulator CDC25B phosphatase in affecting chromosomal activity.
Conclusions:
- Epigenetic modifiers, particularly BET proteins, play a role in EC response to VEGFA blockade.
- Findings offer insights into epigenetic regulation of anti-angiogenesis.
- Potential for developing biomarkers and strategies to overcome AAT resistance.
