Identification of Autophagy-Related Genes as Targets for Senescence Induction Using a Customizable CRISPR-Based Suicide Switch Screen
| العنوان: | Identification of Autophagy-Related Genes as Targets for Senescence Induction Using a Customizable CRISPR-Based Suicide Switch Screen |
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| المؤلفون: | Rodrigo Leite de Oliveira, Giulia De Conti, Cor Lieftink, Arnout Schepers, Roderick L. Beijersbergen, René Bernards, Ziva Pogacar, Fleur Jochems, Liqin Wang |
| المصدر: | Mol Cancer Res |
| بيانات النشر: | American Association for Cancer Research (AACR), 2021. |
| سنة النشر: | 2021 |
| مصطلحات موضوعية: | Senescence, Cancer Research, Autophagy-Related Proteins, Apoptosis, Biology, Article, Cell Line, Small Molecule Libraries, 03 medical and health sciences, 0302 clinical medicine, Cell Line, Tumor, Neoplasms, Autophagy, medicine, Autophagy-Related Protein-1 Homolog, Humans, CRISPR, Senolytic, Molecular Biology, Cellular Senescence, Cell Proliferation, 030304 developmental biology, 0303 health sciences, Cancer, ULK1, medicine.disease, HEK293 Cells, Oncology, A549 Cells, 030220 oncology & carcinogenesis, Cancer cell, Cancer research, CRISPR-Cas Systems |
| الوصف: | 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. |
| تدمد: | 1557-3125 1541-7786 |
| الوصول الحر: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::eab7c3c161e6fd4bc1fdc9b4e06ac955Test https://doi.org/10.1158/1541-7786.mcr-21-0146Test |
| حقوق: | OPEN |
| رقم الانضمام: | edsair.doi.dedup.....eab7c3c161e6fd4bc1fdc9b4e06ac955 |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 15573125 15417786 |
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