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Published on: June 28, 2019
Identification of Autophagy-Related Genes as Targets for Senescence Induction Using a Customizable CRISPR-Based
Arnout Schepers1, Fleur Jochems1, Cor Lieftink1
1Division of Molecular Carcinogenesis, Oncode Institute, Netherlands Cancer Institute, Amsterdam, the Netherlands.
Abstract:
Pro-senescence therapies are increasingly being considered for the treatment of cancer. Identifying additional targets to induce senescence in cancer cells could further enable such therapies. However, screening for targets whose suppression induces senescence on a genome-wide scale is challenging, as senescent cells become growth arrested, and senescence-associated features can take 1 to 2 weeks to develop. For a screen with a whole-genome CRISPR library, this would result in billions of undesirable proliferating cells by the time the senescent features emerge in the growth arrested cells. Here, we present a suicide switch system that allows genome-wide CRISPR screening in growth-arrested subpopulations by eliminating the proliferating cells during the screen through activation of a suicide switch in proliferating cells. Using this system, we identify in a genome-scale CRISPR screen several autophagy-related proteins as targets for senescence induction. We show that inhibiting macroautophagy with a small molecule ULK1 inhibitor can induce senescence in cancer cell lines of different origin. Finally, we show that combining ULK1 inhibition with the senolytic drug ABT-263 leads to apoptosis in a panel of cancer cell lines. IMPLICATIONS: Our suicide switch approach allows for genome-scale identification of pro-senescence targets, and can be adapted to simplify other screens depending on the nature of the promoter used to drive the switch.
Insights
Researchers developed a suicide switch system to identify cancer cell senescence targets. This method enables genome-wide CRISPR screening in growth-arrested cells, revealing autophagy proteins that induce senescence and enhance cancer therapy potential.
Area of Science:
- Oncology
- Cell Biology
- Genetics
Background:
- Pro-senescence therapies are a promising avenue for cancer treatment.
- Identifying novel targets to induce senescence in cancer cells is crucial for advancing these therapies.
- Genome-wide screening for senescence-inducing targets is challenging due to the growth-arrested nature of senescent cells and the prolonged development of senescence-associated features.
Purpose of the Study:
- To develop a novel suicide switch system for genome-wide CRISPR screening in growth-arrested cell subpopulations.
- To identify novel pro-senescence targets by screening a whole-genome CRISPR library.
- To investigate the therapeutic potential of targeting autophagy-related proteins for cancer treatment.
Main Methods:
- Development of a suicide switch system to eliminate proliferating cells during genome-wide CRISPR screening.
- Application of the suicide switch system in a genome-scale CRISPR screen to identify senescence-inducing targets.
- Inhibition of macroautophagy using a small molecule ULK1 inhibitor in various cancer cell lines.
- Combination therapy of ULK1 inhibition with the senolytic drug ABT-263.
Main Results:
- The suicide switch system successfully enabled genome-wide CRISPR screening in growth-arrested subpopulations.
- Several autophagy-related proteins were identified as novel targets for inducing senescence in cancer cells.
- Inhibition of macroautophagy via ULK1 inhibitor induced senescence in diverse cancer cell lines.
- Combined ULK1 inhibition and ABT-263 treatment led to apoptosis in multiple cancer cell lines.
Conclusions:
- The developed suicide switch system is effective for genome-scale identification of pro-senescence targets.
- Targeting autophagy, specifically through ULK1 inhibition, presents a viable strategy for inducing cancer cell senescence.
- Combination therapy with senolytics offers a potential approach for enhancing cancer cell apoptosis.
