Three-dimensional CRISPR screening reveals epigenetic interaction with anti-angiogenic therapy

Michael Y He; Michael M Halford; Ruofei Liu; James P Roy; Zoe L Grant; Leigh Coultas; Niko Thio; Omer Gilan; Yih-Chih Chan; Mark A Dawson; Marc G Achen; Steven A Stacker

doi:10.1038/s42003-021-02397-3

Three-dimensional CRISPR screening reveals epigenetic interaction with anti-angiogenic therapy

Commun Biol. 2021 Jul 15;4(1):878. doi: 10.1038/s42003-021-02397-3.

Authors

Michael Y He^{1

2

3}, Michael M Halford¹, Ruofei Liu^{1

2}, James P Roy^{1

2}, Zoe L Grant^{4

5

6}, Leigh Coultas^{4

5}, Niko Thio⁷, Omer Gilan^{2

8

9}, Yih-Chih Chan⁸, Mark A Dawson^{2

8

10

11}, Marc G Achen^{1

2

12

13}, Steven A Stacker^{14

15

16}

Affiliations

¹ Tumour Angiogenesis and Microenvironment Program, Peter MacCallum Cancer Centre, Melbourne, VIC, Australia.
² Sir Peter MacCallum Department of Oncology, The University of Melbourne, Parkville, VIC, Australia.
³ Princess Margaret Cancer Centre, University Health Network, Toronto, ON, Canada.
⁴ Epigenetics and Development Division, The Walter and Eliza Hall Institute of Medical Research, Parkville, VIC, Australia.
⁵ Department of Medical Biology, The University of Melbourne, Parkville, VIC, Australia.
⁶ Gladstone Institutes, San Francisco, CA, USA.
⁷ Bioinformatics Core, Peter MacCallum Cancer Centre, Melbourne, VIC, Australia.
⁸ Translational Haematology Program, Peter MacCallum Cancer Centre, Melbourne, VIC, Australia.
⁹ Australian Centre for Blood Diseases, Monash University, Melbourne, VIC, Australia.
¹⁰ Centre for Cancer Research, The University of Melbourne, Parkville, VIC, Australia.
¹¹ Department of Haematology, Peter MacCallum Cancer Centre, Melbourne, VIC, Australia.
¹² Department of Surgery, Royal Melbourne Hospital, The University of Melbourne, Parkville, VIC, Australia.
¹³ St Vincent's Institute of Medical Research, Melbourne, VIC, Australia.
¹⁴ Tumour Angiogenesis and Microenvironment Program, Peter MacCallum Cancer Centre, Melbourne, VIC, Australia. [email protected].
¹⁵ Sir Peter MacCallum Department of Oncology, The University of Melbourne, Parkville, VIC, Australia. [email protected].
¹⁶ Department of Surgery, Royal Melbourne Hospital, The University of Melbourne, Parkville, VIC, Australia. [email protected].

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.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Angiogenesis Inhibitors / genetics*
Bevacizumab / genetics*
Blood
Clustered Regularly Interspaced Short Palindromic Repeats*
Endothelial Cells / drug effects
Epigenesis, Genetic*

Substances

Angiogenesis Inhibitors
Bevacizumab