Inhibition of BRD4 attenuated IFNγ-induced apoptosis in colorectal cancer organoids
Abstract Background This study aimed to analyze the functional role of Brd4 in colorectal cancer (CRC) organoids. Brd4 was identified as a CRC-related gene by our previous Sleeping Beauty mutagenesis transposon screening in mice. Brd4 is a transcriptional regulator that recognizes acetylated histone...
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BMC
2025-01-01
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| Series: | BMC Cancer |
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| Online Access: | https://doi.org/10.1186/s12885-025-13544-y |
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| author | Akimi Yonezawa Kana Shimomura Koji Okamoto Haruna Takeda |
| author_facet | Akimi Yonezawa Kana Shimomura Koji Okamoto Haruna Takeda |
| author_sort | Akimi Yonezawa |
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| description | Abstract Background This study aimed to analyze the functional role of Brd4 in colorectal cancer (CRC) organoids. Brd4 was identified as a CRC-related gene by our previous Sleeping Beauty mutagenesis transposon screening in mice. Brd4 is a transcriptional regulator that recognizes acetylated histones and is known to be involved in inflammatory responses. The role of Brd4 in CRC development remains largely unknown. Methods We knocked out Brd4 in tumor organoids carrying mutations in Apc and Kras to generate Brd4KO organoids, and performed RNA-seq. The response of Brd4KO organoids to IFNγ was analyzed via a cell viability assay, an apoptosis assay, and RNAseq. The results were validated by pharmacological inhibition experiments with JQ1 in human CRC organoids. Results In Brd4KO organoids, the IFNγ signaling genes Il33 and Myc target genes were downregulated. The addition of IFNγ to the colon organoids induced apoptosis, but IFNγ-induced apoptosis was attenuated in the Brd4KO organoids compared with the control organoids (two-sided t-test, P < 0.05). Similar results were obtained from pharmacological inhibition with JQ1 in human CRC organoids; IL33 expression was decreased, and IFNγ-induced apoptosis was attenuated in the presence of JQ1. Conclusions Our results showed that the inhibition of Brd4 suppressed IFNγ-induced cytotoxicity by modulating the Jak-Stat pathway. These data suggested that the inhibition of Brd4 could increase cell viability in the cancer microenvironment where IFNγ is abundant, revealing a new aspect of the molecular mechanism of CRC development. Our results may help in evaluating the application of Bet inhibitors in treating CRC. Additionally, our RNA-seq data sets will be helpful for clarifying the relationship between Brd4 and immunomodulators, such as Il33, or for studying the responses of colonic epithelial cells to IFNγ. |
| format | Article |
| id | doaj-art-360977123f08478f9552bcaf11332115 |
| institution | Kabale University |
| issn | 1471-2407 |
| language | English |
| publishDate | 2025-01-01 |
| publisher | BMC |
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| series | BMC Cancer |
| spelling | doaj-art-360977123f08478f9552bcaf113321152025-01-26T12:38:09ZengBMCBMC Cancer1471-24072025-01-012511910.1186/s12885-025-13544-yInhibition of BRD4 attenuated IFNγ-induced apoptosis in colorectal cancer organoidsAkimi Yonezawa0Kana Shimomura1Koji Okamoto2Haruna Takeda3Laboratory of Molecular Genetics, National Cancer Center Research InstituteLaboratory of Molecular Genetics, National Cancer Center Research InstituteAdvanced Comprehensive Research Organization, Teikyo UniversityLaboratory of Molecular Genetics, National Cancer Center Research InstituteAbstract Background This study aimed to analyze the functional role of Brd4 in colorectal cancer (CRC) organoids. Brd4 was identified as a CRC-related gene by our previous Sleeping Beauty mutagenesis transposon screening in mice. Brd4 is a transcriptional regulator that recognizes acetylated histones and is known to be involved in inflammatory responses. The role of Brd4 in CRC development remains largely unknown. Methods We knocked out Brd4 in tumor organoids carrying mutations in Apc and Kras to generate Brd4KO organoids, and performed RNA-seq. The response of Brd4KO organoids to IFNγ was analyzed via a cell viability assay, an apoptosis assay, and RNAseq. The results were validated by pharmacological inhibition experiments with JQ1 in human CRC organoids. Results In Brd4KO organoids, the IFNγ signaling genes Il33 and Myc target genes were downregulated. The addition of IFNγ to the colon organoids induced apoptosis, but IFNγ-induced apoptosis was attenuated in the Brd4KO organoids compared with the control organoids (two-sided t-test, P < 0.05). Similar results were obtained from pharmacological inhibition with JQ1 in human CRC organoids; IL33 expression was decreased, and IFNγ-induced apoptosis was attenuated in the presence of JQ1. Conclusions Our results showed that the inhibition of Brd4 suppressed IFNγ-induced cytotoxicity by modulating the Jak-Stat pathway. These data suggested that the inhibition of Brd4 could increase cell viability in the cancer microenvironment where IFNγ is abundant, revealing a new aspect of the molecular mechanism of CRC development. Our results may help in evaluating the application of Bet inhibitors in treating CRC. Additionally, our RNA-seq data sets will be helpful for clarifying the relationship between Brd4 and immunomodulators, such as Il33, or for studying the responses of colonic epithelial cells to IFNγ.https://doi.org/10.1186/s12885-025-13544-yOrganoidColorectal cancerBrd4ApoptosisJQ1IFNγ |
| spellingShingle | Akimi Yonezawa Kana Shimomura Koji Okamoto Haruna Takeda Inhibition of BRD4 attenuated IFNγ-induced apoptosis in colorectal cancer organoids BMC Cancer Organoid Colorectal cancer Brd4 Apoptosis JQ1 IFNγ |
| title | Inhibition of BRD4 attenuated IFNγ-induced apoptosis in colorectal cancer organoids |
| title_full | Inhibition of BRD4 attenuated IFNγ-induced apoptosis in colorectal cancer organoids |
| title_fullStr | Inhibition of BRD4 attenuated IFNγ-induced apoptosis in colorectal cancer organoids |
| title_full_unstemmed | Inhibition of BRD4 attenuated IFNγ-induced apoptosis in colorectal cancer organoids |
| title_short | Inhibition of BRD4 attenuated IFNγ-induced apoptosis in colorectal cancer organoids |
| title_sort | inhibition of brd4 attenuated ifnγ induced apoptosis in colorectal cancer organoids |
| topic | Organoid Colorectal cancer Brd4 Apoptosis JQ1 IFNγ |
| url | https://doi.org/10.1186/s12885-025-13544-y |
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