Loading…
Synthetic lethality of combined ULK1 defection and p53 restoration induce pyroptosis by directly upregulating GSDME transcription and cleavage activation through ROS/NLRP3 signaling
High expression of ubiquitin ligase MDM2 is a primary cause of p53 inactivation in many tumors, making it a promising therapeutic target. However, MDM2 inhibitors have failed in clinical trials due to p53-induced feedback that enhances MDM2 expression. This underscores the urgent need to find an eff...
Saved in:
Published in: | Journal of experimental & clinical cancer research 2024-08, Vol.43 (1), p.248-20, Article 248 |
---|---|
Main Authors: | , , , , , , , , , , , |
Format: | Article |
Language: | English |
Subjects: | |
Citations: | Items that this one cites |
Online Access: | Get full text |
Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
cited_by | |
---|---|
cites | cdi_FETCH-LOGICAL-c420t-5b33c737dc41da92eb85ad55ca989ddc4694fe13ab81a7dde30de6602f16dcaa3 |
container_end_page | 20 |
container_issue | 1 |
container_start_page | 248 |
container_title | Journal of experimental & clinical cancer research |
container_volume | 43 |
creator | Chen, Wei Yang, Kai-Bin Zhang, Yuan-Zhe Lin, Zai-Shan Chen, Jin-Wei Qi, Si-Fan Wu, Chen-Fei Feng, Gong-Kan Yang, Da-Jun Chen, Ming Zhu, Xiao-Feng Li, Xuan |
description | High expression of ubiquitin ligase MDM2 is a primary cause of p53 inactivation in many tumors, making it a promising therapeutic target. However, MDM2 inhibitors have failed in clinical trials due to p53-induced feedback that enhances MDM2 expression. This underscores the urgent need to find an effective adaptive genotype or combination of targets.
Kinome-wide CRISPR/Cas9 knockout screen was performed to identify genes that modulate the response to MDM2 inhibitor using TP53 wild type cancer cells and found ULK1 as a candidate. The MTT cell viability assay, flow cytometry and LDH assay were conducted to evaluate the activation of pyroptosis and the synthetic lethality effects of combining ULK1 depletion with p53 activation. Dual-luciferase reporter assay and ChIP-qPCR were performed to confirm that p53 directly mediates the transcription of GSDME and to identify the binding region of p53 in the promoter of GSDME. ULK1 knockout / overexpression cells were constructed to investigate the functional role of ULK1 both in vitro and in vivo. The mechanism of ULK1 depletion to activate GSMDE was mainly investigated by qPCR, western blot and ELISA.
By using high-throughput screening, we identified ULK1 as a synthetic lethal gene for the MDM2 inhibitor APG115. It was determined that deletion of ULK1 significantly increased the sensitivity, with cells undergoing typical pyroptosis. Mechanistically, p53 promote pyroptosis initiation by directly mediating GSDME transcription that induce basal-level pyroptosis. Moreover, ULK1 depletion reduces mitophagy, resulting in the accumulation of damaged mitochondria and subsequent increasing of reactive oxygen species (ROS). This in turn cleaves and activates GSDME via the NLRP3-Caspase inflammatory signaling axis. The molecular cascade makes ULK1 act as a crucial regulator of pyroptosis initiation mediated by p53 activation cells. Besides, mitophagy is enhanced in platinum-resistant tumors, and ULK1 depletion/p53 activation has a synergistic lethal effect on these tumors, inducing pyroptosis through GSDME directly.
Our research demonstrates that ULK1 deficiency can synergize with MDM2 inhibitors to induce pyroptosis. p53 plays a direct role in activating GSDME transcription, while ULK1 deficiency triggers upregulation of the ROS-NLRP3 signaling pathway, leading to GSDME cleavage and activation. These findings underscore the pivotal role of p53 in determining pyroptosis and provide new avenues for the clinical application of p53 |
doi_str_mv | 10.1186/s13046-024-03168-8 |
format | article |
fullrecord | <record><control><sourceid>gale_doaj_</sourceid><recordid>TN_cdi_doaj_primary_oai_doaj_org_article_36aa2479810046219abd7234f4f5e8a9</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><galeid>A808377654</galeid><doaj_id>oai_doaj_org_article_36aa2479810046219abd7234f4f5e8a9</doaj_id><sourcerecordid>A808377654</sourcerecordid><originalsourceid>FETCH-LOGICAL-c420t-5b33c737dc41da92eb85ad55ca989ddc4694fe13ab81a7dde30de6602f16dcaa3</originalsourceid><addsrcrecordid>eNptUsFu1DAUjBCIlsIPcECWkBCXtHacOPaxKqVULBR16dl6sZ2sq6yd2kmlfBj_h3e3LC1CPtgazczze2-y7C3Bx4RwdhIJxSXLcVHmmBLGc_4sOyR1xXIhGHv-6H2QvYrxFmNGBBEvswMqClJRVh5mv5azG1dmtAr1ZlxBb8cZ-RYpv26sMxrdLL4SpE1r1Gi9Q-A0GiqKgomjD7DFrNOTMmiYgx9GH21EzYy0DUnSz2gagummPlFdhy6Wn76dozGAiyrYYW-pegP30BkEqcz9znZcBT91K3R9tTz5vrj-QVG0nUs_dN3r7EULfTRvHu6j7Obz-c-zL_ni6uLy7HSRq7LAY141lKqa1lqVRIMoTMMr0FWlQHChE8pE2RpCoeEEaq0NxdowhouWMK0A6FF2ufPVHm7lEOwawiw9WLkFfOgkhDS73kjKAIqyFpzgtJSCCGh0XdCyLdvKcBDJ6-POawj-bkrjk2sblel7cMZPUVIsBMesrMpEff8P9dZPIbWeWAQXFcGU4b-sDlJ961qf5qo2pvKUY07rmm29jv_DSkebtVXemdYm_IngwyPBykA_rqLvp81O4lNisSOq4GMMpt0PiGC5CajcBVSmgMptQCVPoncPrU3N2ui95E8i6W_aE-E9</addsrcrecordid><sourcetype>Open Website</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>3102510360</pqid></control><display><type>article</type><title>Synthetic lethality of combined ULK1 defection and p53 restoration induce pyroptosis by directly upregulating GSDME transcription and cleavage activation through ROS/NLRP3 signaling</title><source>Publicly Available Content Database</source><source>PubMed</source><creator>Chen, Wei ; Yang, Kai-Bin ; Zhang, Yuan-Zhe ; Lin, Zai-Shan ; Chen, Jin-Wei ; Qi, Si-Fan ; Wu, Chen-Fei ; Feng, Gong-Kan ; Yang, Da-Jun ; Chen, Ming ; Zhu, Xiao-Feng ; Li, Xuan</creator><creatorcontrib>Chen, Wei ; Yang, Kai-Bin ; Zhang, Yuan-Zhe ; Lin, Zai-Shan ; Chen, Jin-Wei ; Qi, Si-Fan ; Wu, Chen-Fei ; Feng, Gong-Kan ; Yang, Da-Jun ; Chen, Ming ; Zhu, Xiao-Feng ; Li, Xuan</creatorcontrib><description>High expression of ubiquitin ligase MDM2 is a primary cause of p53 inactivation in many tumors, making it a promising therapeutic target. However, MDM2 inhibitors have failed in clinical trials due to p53-induced feedback that enhances MDM2 expression. This underscores the urgent need to find an effective adaptive genotype or combination of targets.
Kinome-wide CRISPR/Cas9 knockout screen was performed to identify genes that modulate the response to MDM2 inhibitor using TP53 wild type cancer cells and found ULK1 as a candidate. The MTT cell viability assay, flow cytometry and LDH assay were conducted to evaluate the activation of pyroptosis and the synthetic lethality effects of combining ULK1 depletion with p53 activation. Dual-luciferase reporter assay and ChIP-qPCR were performed to confirm that p53 directly mediates the transcription of GSDME and to identify the binding region of p53 in the promoter of GSDME. ULK1 knockout / overexpression cells were constructed to investigate the functional role of ULK1 both in vitro and in vivo. The mechanism of ULK1 depletion to activate GSMDE was mainly investigated by qPCR, western blot and ELISA.
By using high-throughput screening, we identified ULK1 as a synthetic lethal gene for the MDM2 inhibitor APG115. It was determined that deletion of ULK1 significantly increased the sensitivity, with cells undergoing typical pyroptosis. Mechanistically, p53 promote pyroptosis initiation by directly mediating GSDME transcription that induce basal-level pyroptosis. Moreover, ULK1 depletion reduces mitophagy, resulting in the accumulation of damaged mitochondria and subsequent increasing of reactive oxygen species (ROS). This in turn cleaves and activates GSDME via the NLRP3-Caspase inflammatory signaling axis. The molecular cascade makes ULK1 act as a crucial regulator of pyroptosis initiation mediated by p53 activation cells. Besides, mitophagy is enhanced in platinum-resistant tumors, and ULK1 depletion/p53 activation has a synergistic lethal effect on these tumors, inducing pyroptosis through GSDME directly.
Our research demonstrates that ULK1 deficiency can synergize with MDM2 inhibitors to induce pyroptosis. p53 plays a direct role in activating GSDME transcription, while ULK1 deficiency triggers upregulation of the ROS-NLRP3 signaling pathway, leading to GSDME cleavage and activation. These findings underscore the pivotal role of p53 in determining pyroptosis and provide new avenues for the clinical application of p53 restoration therapies, as well as suggesting potential combination strategies.</description><identifier>ISSN: 1756-9966</identifier><identifier>ISSN: 0392-9078</identifier><identifier>EISSN: 1756-9966</identifier><identifier>DOI: 10.1186/s13046-024-03168-8</identifier><identifier>PMID: 39215364</identifier><language>eng</language><publisher>England: BioMed Central Ltd</publisher><subject>Animals ; Antibodies ; Autophagy-Related Protein-1 Homolog - genetics ; Autophagy-Related Protein-1 Homolog - metabolism ; Cancer ; Cell culture ; Cell cycle ; Cell death ; Cell Line, Tumor ; Cloning ; CRISPR ; DNA methylation ; Enzyme-linked immunosorbent assay ; Enzymes ; Ethylenediaminetetraacetic acid ; Female ; Genes ; Genetic aspects ; Genetic transcription ; Health aspects ; Humans ; Intracellular Signaling Peptides and Proteins - genetics ; Intracellular Signaling Peptides and Proteins - metabolism ; Kinases ; Ligases ; MDM2 inhibitor ; Mice ; Mitophagy ; NLR Family, Pyrin Domain-Containing 3 Protein ; Penicillin ; Plasmids ; Proteins ; Pyroptosis ; Reactive oxygen species ; Reactive Oxygen Species - metabolism ; Signal Transduction ; Synthetic Lethal Mutations ; TP53 ; Tumor proteins ; Tumor Suppressor Protein p53 - genetics ; Tumor Suppressor Protein p53 - metabolism ; Tumors ; Ubiquitin ; ULK1 ; Up-Regulation</subject><ispartof>Journal of experimental & clinical cancer research, 2024-08, Vol.43 (1), p.248-20, Article 248</ispartof><rights>2024. The Author(s).</rights><rights>COPYRIGHT 2024 BioMed Central Ltd.</rights><rights>2024. This work is licensed under http://creativecommons.org/licenses/by/4.0/ (the “License”). Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><cites>FETCH-LOGICAL-c420t-5b33c737dc41da92eb85ad55ca989ddc4694fe13ab81a7dde30de6602f16dcaa3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://www.proquest.com/docview/3102510360?pq-origsite=primo$$EHTML$$P50$$Gproquest$$Hfree_for_read</linktohtml><link.rule.ids>314,780,784,25753,27924,27925,37012,37013,44590</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/39215364$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Chen, Wei</creatorcontrib><creatorcontrib>Yang, Kai-Bin</creatorcontrib><creatorcontrib>Zhang, Yuan-Zhe</creatorcontrib><creatorcontrib>Lin, Zai-Shan</creatorcontrib><creatorcontrib>Chen, Jin-Wei</creatorcontrib><creatorcontrib>Qi, Si-Fan</creatorcontrib><creatorcontrib>Wu, Chen-Fei</creatorcontrib><creatorcontrib>Feng, Gong-Kan</creatorcontrib><creatorcontrib>Yang, Da-Jun</creatorcontrib><creatorcontrib>Chen, Ming</creatorcontrib><creatorcontrib>Zhu, Xiao-Feng</creatorcontrib><creatorcontrib>Li, Xuan</creatorcontrib><title>Synthetic lethality of combined ULK1 defection and p53 restoration induce pyroptosis by directly upregulating GSDME transcription and cleavage activation through ROS/NLRP3 signaling</title><title>Journal of experimental & clinical cancer research</title><addtitle>J Exp Clin Cancer Res</addtitle><description>High expression of ubiquitin ligase MDM2 is a primary cause of p53 inactivation in many tumors, making it a promising therapeutic target. However, MDM2 inhibitors have failed in clinical trials due to p53-induced feedback that enhances MDM2 expression. This underscores the urgent need to find an effective adaptive genotype or combination of targets.
Kinome-wide CRISPR/Cas9 knockout screen was performed to identify genes that modulate the response to MDM2 inhibitor using TP53 wild type cancer cells and found ULK1 as a candidate. The MTT cell viability assay, flow cytometry and LDH assay were conducted to evaluate the activation of pyroptosis and the synthetic lethality effects of combining ULK1 depletion with p53 activation. Dual-luciferase reporter assay and ChIP-qPCR were performed to confirm that p53 directly mediates the transcription of GSDME and to identify the binding region of p53 in the promoter of GSDME. ULK1 knockout / overexpression cells were constructed to investigate the functional role of ULK1 both in vitro and in vivo. The mechanism of ULK1 depletion to activate GSMDE was mainly investigated by qPCR, western blot and ELISA.
By using high-throughput screening, we identified ULK1 as a synthetic lethal gene for the MDM2 inhibitor APG115. It was determined that deletion of ULK1 significantly increased the sensitivity, with cells undergoing typical pyroptosis. Mechanistically, p53 promote pyroptosis initiation by directly mediating GSDME transcription that induce basal-level pyroptosis. Moreover, ULK1 depletion reduces mitophagy, resulting in the accumulation of damaged mitochondria and subsequent increasing of reactive oxygen species (ROS). This in turn cleaves and activates GSDME via the NLRP3-Caspase inflammatory signaling axis. The molecular cascade makes ULK1 act as a crucial regulator of pyroptosis initiation mediated by p53 activation cells. Besides, mitophagy is enhanced in platinum-resistant tumors, and ULK1 depletion/p53 activation has a synergistic lethal effect on these tumors, inducing pyroptosis through GSDME directly.
Our research demonstrates that ULK1 deficiency can synergize with MDM2 inhibitors to induce pyroptosis. p53 plays a direct role in activating GSDME transcription, while ULK1 deficiency triggers upregulation of the ROS-NLRP3 signaling pathway, leading to GSDME cleavage and activation. These findings underscore the pivotal role of p53 in determining pyroptosis and provide new avenues for the clinical application of p53 restoration therapies, as well as suggesting potential combination strategies.</description><subject>Animals</subject><subject>Antibodies</subject><subject>Autophagy-Related Protein-1 Homolog - genetics</subject><subject>Autophagy-Related Protein-1 Homolog - metabolism</subject><subject>Cancer</subject><subject>Cell culture</subject><subject>Cell cycle</subject><subject>Cell death</subject><subject>Cell Line, Tumor</subject><subject>Cloning</subject><subject>CRISPR</subject><subject>DNA methylation</subject><subject>Enzyme-linked immunosorbent assay</subject><subject>Enzymes</subject><subject>Ethylenediaminetetraacetic acid</subject><subject>Female</subject><subject>Genes</subject><subject>Genetic aspects</subject><subject>Genetic transcription</subject><subject>Health aspects</subject><subject>Humans</subject><subject>Intracellular Signaling Peptides and Proteins - genetics</subject><subject>Intracellular Signaling Peptides and Proteins - metabolism</subject><subject>Kinases</subject><subject>Ligases</subject><subject>MDM2 inhibitor</subject><subject>Mice</subject><subject>Mitophagy</subject><subject>NLR Family, Pyrin Domain-Containing 3 Protein</subject><subject>Penicillin</subject><subject>Plasmids</subject><subject>Proteins</subject><subject>Pyroptosis</subject><subject>Reactive oxygen species</subject><subject>Reactive Oxygen Species - metabolism</subject><subject>Signal Transduction</subject><subject>Synthetic Lethal Mutations</subject><subject>TP53</subject><subject>Tumor proteins</subject><subject>Tumor Suppressor Protein p53 - genetics</subject><subject>Tumor Suppressor Protein p53 - metabolism</subject><subject>Tumors</subject><subject>Ubiquitin</subject><subject>ULK1</subject><subject>Up-Regulation</subject><issn>1756-9966</issn><issn>0392-9078</issn><issn>1756-9966</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2024</creationdate><recordtype>article</recordtype><sourceid>PIMPY</sourceid><sourceid>DOA</sourceid><recordid>eNptUsFu1DAUjBCIlsIPcECWkBCXtHacOPaxKqVULBR16dl6sZ2sq6yd2kmlfBj_h3e3LC1CPtgazczze2-y7C3Bx4RwdhIJxSXLcVHmmBLGc_4sOyR1xXIhGHv-6H2QvYrxFmNGBBEvswMqClJRVh5mv5azG1dmtAr1ZlxBb8cZ-RYpv26sMxrdLL4SpE1r1Gi9Q-A0GiqKgomjD7DFrNOTMmiYgx9GH21EzYy0DUnSz2gagummPlFdhy6Wn76dozGAiyrYYW-pegP30BkEqcz9znZcBT91K3R9tTz5vrj-QVG0nUs_dN3r7EULfTRvHu6j7Obz-c-zL_ni6uLy7HSRq7LAY141lKqa1lqVRIMoTMMr0FWlQHChE8pE2RpCoeEEaq0NxdowhouWMK0A6FF2ufPVHm7lEOwawiw9WLkFfOgkhDS73kjKAIqyFpzgtJSCCGh0XdCyLdvKcBDJ6-POawj-bkrjk2sblel7cMZPUVIsBMesrMpEff8P9dZPIbWeWAQXFcGU4b-sDlJ961qf5qo2pvKUY07rmm29jv_DSkebtVXemdYm_IngwyPBykA_rqLvp81O4lNisSOq4GMMpt0PiGC5CajcBVSmgMptQCVPoncPrU3N2ui95E8i6W_aE-E9</recordid><startdate>20240830</startdate><enddate>20240830</enddate><creator>Chen, Wei</creator><creator>Yang, Kai-Bin</creator><creator>Zhang, Yuan-Zhe</creator><creator>Lin, Zai-Shan</creator><creator>Chen, Jin-Wei</creator><creator>Qi, Si-Fan</creator><creator>Wu, Chen-Fei</creator><creator>Feng, Gong-Kan</creator><creator>Yang, Da-Jun</creator><creator>Chen, Ming</creator><creator>Zhu, Xiao-Feng</creator><creator>Li, Xuan</creator><general>BioMed Central Ltd</general><general>BioMed Central</general><general>BMC</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>3V.</scope><scope>7X7</scope><scope>7XB</scope><scope>88E</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>ABUWG</scope><scope>AFKRA</scope><scope>AZQEC</scope><scope>BENPR</scope><scope>CCPQU</scope><scope>DWQXO</scope><scope>FYUFA</scope><scope>GHDGH</scope><scope>K9.</scope><scope>M0S</scope><scope>M1P</scope><scope>PIMPY</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>PRINS</scope><scope>7X8</scope><scope>DOA</scope></search><sort><creationdate>20240830</creationdate><title>Synthetic lethality of combined ULK1 defection and p53 restoration induce pyroptosis by directly upregulating GSDME transcription and cleavage activation through ROS/NLRP3 signaling</title><author>Chen, Wei ; Yang, Kai-Bin ; Zhang, Yuan-Zhe ; Lin, Zai-Shan ; Chen, Jin-Wei ; Qi, Si-Fan ; Wu, Chen-Fei ; Feng, Gong-Kan ; Yang, Da-Jun ; Chen, Ming ; Zhu, Xiao-Feng ; Li, Xuan</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c420t-5b33c737dc41da92eb85ad55ca989ddc4694fe13ab81a7dde30de6602f16dcaa3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2024</creationdate><topic>Animals</topic><topic>Antibodies</topic><topic>Autophagy-Related Protein-1 Homolog - genetics</topic><topic>Autophagy-Related Protein-1 Homolog - metabolism</topic><topic>Cancer</topic><topic>Cell culture</topic><topic>Cell cycle</topic><topic>Cell death</topic><topic>Cell Line, Tumor</topic><topic>Cloning</topic><topic>CRISPR</topic><topic>DNA methylation</topic><topic>Enzyme-linked immunosorbent assay</topic><topic>Enzymes</topic><topic>Ethylenediaminetetraacetic acid</topic><topic>Female</topic><topic>Genes</topic><topic>Genetic aspects</topic><topic>Genetic transcription</topic><topic>Health aspects</topic><topic>Humans</topic><topic>Intracellular Signaling Peptides and Proteins - genetics</topic><topic>Intracellular Signaling Peptides and Proteins - metabolism</topic><topic>Kinases</topic><topic>Ligases</topic><topic>MDM2 inhibitor</topic><topic>Mice</topic><topic>Mitophagy</topic><topic>NLR Family, Pyrin Domain-Containing 3 Protein</topic><topic>Penicillin</topic><topic>Plasmids</topic><topic>Proteins</topic><topic>Pyroptosis</topic><topic>Reactive oxygen species</topic><topic>Reactive Oxygen Species - metabolism</topic><topic>Signal Transduction</topic><topic>Synthetic Lethal Mutations</topic><topic>TP53</topic><topic>Tumor proteins</topic><topic>Tumor Suppressor Protein p53 - genetics</topic><topic>Tumor Suppressor Protein p53 - metabolism</topic><topic>Tumors</topic><topic>Ubiquitin</topic><topic>ULK1</topic><topic>Up-Regulation</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Chen, Wei</creatorcontrib><creatorcontrib>Yang, Kai-Bin</creatorcontrib><creatorcontrib>Zhang, Yuan-Zhe</creatorcontrib><creatorcontrib>Lin, Zai-Shan</creatorcontrib><creatorcontrib>Chen, Jin-Wei</creatorcontrib><creatorcontrib>Qi, Si-Fan</creatorcontrib><creatorcontrib>Wu, Chen-Fei</creatorcontrib><creatorcontrib>Feng, Gong-Kan</creatorcontrib><creatorcontrib>Yang, Da-Jun</creatorcontrib><creatorcontrib>Chen, Ming</creatorcontrib><creatorcontrib>Zhu, Xiao-Feng</creatorcontrib><creatorcontrib>Li, Xuan</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>ProQuest Central (Corporate)</collection><collection>Health & Medical Collection</collection><collection>ProQuest Central (purchase pre-March 2016)</collection><collection>Medical Database (Alumni Edition)</collection><collection>Hospital Premium Collection</collection><collection>Hospital Premium Collection (Alumni Edition)</collection><collection>ProQuest Central (Alumni) (purchase pre-March 2016)</collection><collection>ProQuest Central (Alumni)</collection><collection>ProQuest Central</collection><collection>ProQuest Central Essentials</collection><collection>ProQuest Central</collection><collection>ProQuest One Community College</collection><collection>ProQuest Central Korea</collection><collection>Health Research Premium Collection</collection><collection>Health Research Premium Collection (Alumni)</collection><collection>ProQuest Health & Medical Complete (Alumni)</collection><collection>Health & Medical Collection (Alumni Edition)</collection><collection>PML(ProQuest Medical Library)</collection><collection>Publicly Available Content Database</collection><collection>ProQuest One Academic Eastern Edition (DO NOT USE)</collection><collection>ProQuest One Academic</collection><collection>ProQuest One Academic UKI Edition</collection><collection>ProQuest Central China</collection><collection>MEDLINE - Academic</collection><collection>Directory of Open Access Journals</collection><jtitle>Journal of experimental & clinical cancer research</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Chen, Wei</au><au>Yang, Kai-Bin</au><au>Zhang, Yuan-Zhe</au><au>Lin, Zai-Shan</au><au>Chen, Jin-Wei</au><au>Qi, Si-Fan</au><au>Wu, Chen-Fei</au><au>Feng, Gong-Kan</au><au>Yang, Da-Jun</au><au>Chen, Ming</au><au>Zhu, Xiao-Feng</au><au>Li, Xuan</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Synthetic lethality of combined ULK1 defection and p53 restoration induce pyroptosis by directly upregulating GSDME transcription and cleavage activation through ROS/NLRP3 signaling</atitle><jtitle>Journal of experimental & clinical cancer research</jtitle><addtitle>J Exp Clin Cancer Res</addtitle><date>2024-08-30</date><risdate>2024</risdate><volume>43</volume><issue>1</issue><spage>248</spage><epage>20</epage><pages>248-20</pages><artnum>248</artnum><issn>1756-9966</issn><issn>0392-9078</issn><eissn>1756-9966</eissn><abstract>High expression of ubiquitin ligase MDM2 is a primary cause of p53 inactivation in many tumors, making it a promising therapeutic target. However, MDM2 inhibitors have failed in clinical trials due to p53-induced feedback that enhances MDM2 expression. This underscores the urgent need to find an effective adaptive genotype or combination of targets.
Kinome-wide CRISPR/Cas9 knockout screen was performed to identify genes that modulate the response to MDM2 inhibitor using TP53 wild type cancer cells and found ULK1 as a candidate. The MTT cell viability assay, flow cytometry and LDH assay were conducted to evaluate the activation of pyroptosis and the synthetic lethality effects of combining ULK1 depletion with p53 activation. Dual-luciferase reporter assay and ChIP-qPCR were performed to confirm that p53 directly mediates the transcription of GSDME and to identify the binding region of p53 in the promoter of GSDME. ULK1 knockout / overexpression cells were constructed to investigate the functional role of ULK1 both in vitro and in vivo. The mechanism of ULK1 depletion to activate GSMDE was mainly investigated by qPCR, western blot and ELISA.
By using high-throughput screening, we identified ULK1 as a synthetic lethal gene for the MDM2 inhibitor APG115. It was determined that deletion of ULK1 significantly increased the sensitivity, with cells undergoing typical pyroptosis. Mechanistically, p53 promote pyroptosis initiation by directly mediating GSDME transcription that induce basal-level pyroptosis. Moreover, ULK1 depletion reduces mitophagy, resulting in the accumulation of damaged mitochondria and subsequent increasing of reactive oxygen species (ROS). This in turn cleaves and activates GSDME via the NLRP3-Caspase inflammatory signaling axis. The molecular cascade makes ULK1 act as a crucial regulator of pyroptosis initiation mediated by p53 activation cells. Besides, mitophagy is enhanced in platinum-resistant tumors, and ULK1 depletion/p53 activation has a synergistic lethal effect on these tumors, inducing pyroptosis through GSDME directly.
Our research demonstrates that ULK1 deficiency can synergize with MDM2 inhibitors to induce pyroptosis. p53 plays a direct role in activating GSDME transcription, while ULK1 deficiency triggers upregulation of the ROS-NLRP3 signaling pathway, leading to GSDME cleavage and activation. These findings underscore the pivotal role of p53 in determining pyroptosis and provide new avenues for the clinical application of p53 restoration therapies, as well as suggesting potential combination strategies.</abstract><cop>England</cop><pub>BioMed Central Ltd</pub><pmid>39215364</pmid><doi>10.1186/s13046-024-03168-8</doi><tpages>20</tpages><oa>free_for_read</oa></addata></record> |
fulltext | fulltext |
identifier | ISSN: 1756-9966 |
ispartof | Journal of experimental & clinical cancer research, 2024-08, Vol.43 (1), p.248-20, Article 248 |
issn | 1756-9966 0392-9078 1756-9966 |
language | eng |
recordid | cdi_doaj_primary_oai_doaj_org_article_36aa2479810046219abd7234f4f5e8a9 |
source | Publicly Available Content Database; PubMed |
subjects | Animals Antibodies Autophagy-Related Protein-1 Homolog - genetics Autophagy-Related Protein-1 Homolog - metabolism Cancer Cell culture Cell cycle Cell death Cell Line, Tumor Cloning CRISPR DNA methylation Enzyme-linked immunosorbent assay Enzymes Ethylenediaminetetraacetic acid Female Genes Genetic aspects Genetic transcription Health aspects Humans Intracellular Signaling Peptides and Proteins - genetics Intracellular Signaling Peptides and Proteins - metabolism Kinases Ligases MDM2 inhibitor Mice Mitophagy NLR Family, Pyrin Domain-Containing 3 Protein Penicillin Plasmids Proteins Pyroptosis Reactive oxygen species Reactive Oxygen Species - metabolism Signal Transduction Synthetic Lethal Mutations TP53 Tumor proteins Tumor Suppressor Protein p53 - genetics Tumor Suppressor Protein p53 - metabolism Tumors Ubiquitin ULK1 Up-Regulation |
title | Synthetic lethality of combined ULK1 defection and p53 restoration induce pyroptosis by directly upregulating GSDME transcription and cleavage activation through ROS/NLRP3 signaling |
url | http://sfxeu10.hosted.exlibrisgroup.com/loughborough?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2024-12-23T23%3A54%3A19IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-gale_doaj_&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=Synthetic%20lethality%20of%20combined%20ULK1%20defection%20and%20p53%20restoration%20induce%20pyroptosis%20by%20directly%20upregulating%20GSDME%20transcription%20and%20cleavage%20activation%20through%20ROS/NLRP3%20signaling&rft.jtitle=Journal%20of%20experimental%20&%20clinical%20cancer%20research&rft.au=Chen,%20Wei&rft.date=2024-08-30&rft.volume=43&rft.issue=1&rft.spage=248&rft.epage=20&rft.pages=248-20&rft.artnum=248&rft.issn=1756-9966&rft.eissn=1756-9966&rft_id=info:doi/10.1186/s13046-024-03168-8&rft_dat=%3Cgale_doaj_%3EA808377654%3C/gale_doaj_%3E%3Cgrp_id%3Ecdi_FETCH-LOGICAL-c420t-5b33c737dc41da92eb85ad55ca989ddc4694fe13ab81a7dde30de6602f16dcaa3%3C/grp_id%3E%3Coa%3E%3C/oa%3E%3Curl%3E%3C/url%3E&rft_id=info:oai/&rft_pqid=3102510360&rft_id=info:pmid/39215364&rft_galeid=A808377654&rfr_iscdi=true |