A 'synthetic-sickness' screen for senescence re-engagement targets in mutant cancer backgrounds.
Senescence is a universal barrier to immortalisation and tumorigenesis. As such, interest in the use of senescence-induction in a therapeutic context has been gaining momentum in the past few years; however, senescence and immortalisation remain underserved areas for drug discovery owing to a lack o...
Saved in:
| Main Authors: | , , , , , , , , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Public Library of Science (PLoS)
2017-08-01
|
| Series: | PLoS Genetics |
| Online Access: | https://journals.plos.org/plosgenetics/article/file?id=10.1371/journal.pgen.1006942&type=printable |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1849715598309720064 |
|---|---|
| author | Claire J Cairney Lauren S Godwin Alan E Bilsland Sharon Burns Katrina H Stevenson Lynn McGarry John Revie Jon D Moore Ceri M Wiggins Rebecca S Collinson Clare Mudd Elpida Tsonou Mahito Sadaie Dorothy C Bennett Masashi Narita Christopher J Torrance W Nicol Keith |
| author_facet | Claire J Cairney Lauren S Godwin Alan E Bilsland Sharon Burns Katrina H Stevenson Lynn McGarry John Revie Jon D Moore Ceri M Wiggins Rebecca S Collinson Clare Mudd Elpida Tsonou Mahito Sadaie Dorothy C Bennett Masashi Narita Christopher J Torrance W Nicol Keith |
| author_sort | Claire J Cairney |
| collection | DOAJ |
| description | Senescence is a universal barrier to immortalisation and tumorigenesis. As such, interest in the use of senescence-induction in a therapeutic context has been gaining momentum in the past few years; however, senescence and immortalisation remain underserved areas for drug discovery owing to a lack of robust senescence inducing agents and an incomplete understanding of the signalling events underlying this complex process. In order to address this issue we undertook a large-scale morphological siRNA screen for inducers of senescence phenotypes in the human melanoma cell line A375P. Following rescreen and validation in a second cancer cell line, HCT116 colorectal carcinoma, a panel of 16 of the most robust hits were selected for further validation based on significance and the potential to be targeted by drug-like molecules. Using secondary assays for detection of senescence biomarkers p21, 53BP1 and senescence associated beta-galactosidase (SAβGal) in a panel of HCT116 cell lines carrying cancer-relevant mutations, we show that partial senescence phenotypes can be induced to varying degrees in a context dependent manner, even in the absence of p21 or p53 expression. However, proliferation arrest varied among genetic backgrounds with predominantly toxic effects in p21 null cells, while cells lacking PI3K mutation failed to arrest. Furthermore, we show that the oncogene ECT2 induces partial senescence phenotypes in all mutant backgrounds tested, demonstrating a dependence on activating KRASG13D for growth suppression and a complete senescence response. These results suggest a potential mechanism to target mutant KRAS signalling through ECT2 in cancers that are reliant on activating KRAS mutations and remain refractory to current treatments. |
| format | Article |
| id | doaj-art-9c37fdc4728e4b2584673b3d2d9b5b8b |
| institution | DOAJ |
| issn | 1553-7390 1553-7404 |
| language | English |
| publishDate | 2017-08-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS Genetics |
| spelling | doaj-art-9c37fdc4728e4b2584673b3d2d9b5b8b2025-08-20T03:13:18ZengPublic Library of Science (PLoS)PLoS Genetics1553-73901553-74042017-08-01138e100694210.1371/journal.pgen.1006942A 'synthetic-sickness' screen for senescence re-engagement targets in mutant cancer backgrounds.Claire J CairneyLauren S GodwinAlan E BilslandSharon BurnsKatrina H StevensonLynn McGarryJohn RevieJon D MooreCeri M WigginsRebecca S CollinsonClare MuddElpida TsonouMahito SadaieDorothy C BennettMasashi NaritaChristopher J TorranceW Nicol KeithSenescence is a universal barrier to immortalisation and tumorigenesis. As such, interest in the use of senescence-induction in a therapeutic context has been gaining momentum in the past few years; however, senescence and immortalisation remain underserved areas for drug discovery owing to a lack of robust senescence inducing agents and an incomplete understanding of the signalling events underlying this complex process. In order to address this issue we undertook a large-scale morphological siRNA screen for inducers of senescence phenotypes in the human melanoma cell line A375P. Following rescreen and validation in a second cancer cell line, HCT116 colorectal carcinoma, a panel of 16 of the most robust hits were selected for further validation based on significance and the potential to be targeted by drug-like molecules. Using secondary assays for detection of senescence biomarkers p21, 53BP1 and senescence associated beta-galactosidase (SAβGal) in a panel of HCT116 cell lines carrying cancer-relevant mutations, we show that partial senescence phenotypes can be induced to varying degrees in a context dependent manner, even in the absence of p21 or p53 expression. However, proliferation arrest varied among genetic backgrounds with predominantly toxic effects in p21 null cells, while cells lacking PI3K mutation failed to arrest. Furthermore, we show that the oncogene ECT2 induces partial senescence phenotypes in all mutant backgrounds tested, demonstrating a dependence on activating KRASG13D for growth suppression and a complete senescence response. These results suggest a potential mechanism to target mutant KRAS signalling through ECT2 in cancers that are reliant on activating KRAS mutations and remain refractory to current treatments.https://journals.plos.org/plosgenetics/article/file?id=10.1371/journal.pgen.1006942&type=printable |
| spellingShingle | Claire J Cairney Lauren S Godwin Alan E Bilsland Sharon Burns Katrina H Stevenson Lynn McGarry John Revie Jon D Moore Ceri M Wiggins Rebecca S Collinson Clare Mudd Elpida Tsonou Mahito Sadaie Dorothy C Bennett Masashi Narita Christopher J Torrance W Nicol Keith A 'synthetic-sickness' screen for senescence re-engagement targets in mutant cancer backgrounds. PLoS Genetics |
| title | A 'synthetic-sickness' screen for senescence re-engagement targets in mutant cancer backgrounds. |
| title_full | A 'synthetic-sickness' screen for senescence re-engagement targets in mutant cancer backgrounds. |
| title_fullStr | A 'synthetic-sickness' screen for senescence re-engagement targets in mutant cancer backgrounds. |
| title_full_unstemmed | A 'synthetic-sickness' screen for senescence re-engagement targets in mutant cancer backgrounds. |
| title_short | A 'synthetic-sickness' screen for senescence re-engagement targets in mutant cancer backgrounds. |
| title_sort | synthetic sickness screen for senescence re engagement targets in mutant cancer backgrounds |
| url | https://journals.plos.org/plosgenetics/article/file?id=10.1371/journal.pgen.1006942&type=printable |
| work_keys_str_mv | AT clairejcairney asyntheticsicknessscreenforsenescencereengagementtargetsinmutantcancerbackgrounds AT laurensgodwin asyntheticsicknessscreenforsenescencereengagementtargetsinmutantcancerbackgrounds AT alanebilsland asyntheticsicknessscreenforsenescencereengagementtargetsinmutantcancerbackgrounds AT sharonburns asyntheticsicknessscreenforsenescencereengagementtargetsinmutantcancerbackgrounds AT katrinahstevenson asyntheticsicknessscreenforsenescencereengagementtargetsinmutantcancerbackgrounds AT lynnmcgarry asyntheticsicknessscreenforsenescencereengagementtargetsinmutantcancerbackgrounds AT johnrevie asyntheticsicknessscreenforsenescencereengagementtargetsinmutantcancerbackgrounds AT jondmoore asyntheticsicknessscreenforsenescencereengagementtargetsinmutantcancerbackgrounds AT cerimwiggins asyntheticsicknessscreenforsenescencereengagementtargetsinmutantcancerbackgrounds AT rebeccascollinson asyntheticsicknessscreenforsenescencereengagementtargetsinmutantcancerbackgrounds AT claremudd asyntheticsicknessscreenforsenescencereengagementtargetsinmutantcancerbackgrounds AT elpidatsonou asyntheticsicknessscreenforsenescencereengagementtargetsinmutantcancerbackgrounds AT mahitosadaie asyntheticsicknessscreenforsenescencereengagementtargetsinmutantcancerbackgrounds AT dorothycbennett asyntheticsicknessscreenforsenescencereengagementtargetsinmutantcancerbackgrounds AT masashinarita asyntheticsicknessscreenforsenescencereengagementtargetsinmutantcancerbackgrounds AT christopherjtorrance asyntheticsicknessscreenforsenescencereengagementtargetsinmutantcancerbackgrounds AT wnicolkeith asyntheticsicknessscreenforsenescencereengagementtargetsinmutantcancerbackgrounds AT clairejcairney syntheticsicknessscreenforsenescencereengagementtargetsinmutantcancerbackgrounds AT laurensgodwin syntheticsicknessscreenforsenescencereengagementtargetsinmutantcancerbackgrounds AT alanebilsland syntheticsicknessscreenforsenescencereengagementtargetsinmutantcancerbackgrounds AT sharonburns syntheticsicknessscreenforsenescencereengagementtargetsinmutantcancerbackgrounds AT katrinahstevenson syntheticsicknessscreenforsenescencereengagementtargetsinmutantcancerbackgrounds AT lynnmcgarry syntheticsicknessscreenforsenescencereengagementtargetsinmutantcancerbackgrounds AT johnrevie syntheticsicknessscreenforsenescencereengagementtargetsinmutantcancerbackgrounds AT jondmoore syntheticsicknessscreenforsenescencereengagementtargetsinmutantcancerbackgrounds AT cerimwiggins syntheticsicknessscreenforsenescencereengagementtargetsinmutantcancerbackgrounds AT rebeccascollinson syntheticsicknessscreenforsenescencereengagementtargetsinmutantcancerbackgrounds AT claremudd syntheticsicknessscreenforsenescencereengagementtargetsinmutantcancerbackgrounds AT elpidatsonou syntheticsicknessscreenforsenescencereengagementtargetsinmutantcancerbackgrounds AT mahitosadaie syntheticsicknessscreenforsenescencereengagementtargetsinmutantcancerbackgrounds AT dorothycbennett syntheticsicknessscreenforsenescencereengagementtargetsinmutantcancerbackgrounds AT masashinarita syntheticsicknessscreenforsenescencereengagementtargetsinmutantcancerbackgrounds AT christopherjtorrance syntheticsicknessscreenforsenescencereengagementtargetsinmutantcancerbackgrounds AT wnicolkeith syntheticsicknessscreenforsenescencereengagementtargetsinmutantcancerbackgrounds |