Transcriptomic profiling reveals RetS-mediated regulation of type VI secretion system and host cell responses in Pseudomonas aeruginosa infections
Pseudomonas aeruginosa is a major opportunistic pathogen that causes chronic infections, particularly in patients with cystic fibrosis and chronic obstructive pulmonary disease (COPD). The type VI secretion system (T6SS) is a primary virulence factor of P. aeruginosa in chronic infections. The objec...
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Frontiers Media S.A.
2025-06-01
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| Series: | Frontiers in Cellular and Infection Microbiology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fcimb.2025.1582339/full |
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| author | Yinglin Wu Yinglin Wu Yinglin Wu Shan Huang Shan Huang Shan Huang Kai Zhang Yangcheng Shen Yangcheng Shen Yangcheng Shen Shebin Zhang Shebin Zhang Shebin Zhang Haining Xia Haining Xia Haining Xia Jieying Pu Jieying Pu Jieying Pu Cong Shen Cong Shen Cong Shen Cha Chen Cha Chen Cha Chen Jianming Zeng Jianming Zeng Jianming Zeng |
| author_facet | Yinglin Wu Yinglin Wu Yinglin Wu Shan Huang Shan Huang Shan Huang Kai Zhang Yangcheng Shen Yangcheng Shen Yangcheng Shen Shebin Zhang Shebin Zhang Shebin Zhang Haining Xia Haining Xia Haining Xia Jieying Pu Jieying Pu Jieying Pu Cong Shen Cong Shen Cong Shen Cha Chen Cha Chen Cha Chen Jianming Zeng Jianming Zeng Jianming Zeng |
| author_sort | Yinglin Wu |
| collection | DOAJ |
| description | Pseudomonas aeruginosa is a major opportunistic pathogen that causes chronic infections, particularly in patients with cystic fibrosis and chronic obstructive pulmonary disease (COPD). The type VI secretion system (T6SS) is a primary virulence factor of P. aeruginosa in chronic infections. The objective of this study was to elucidate the regulatory mechanisms and pathogenic effects of the T6SS during P. aeruginosa infection, utilizing transcriptome sequencing and functional assays. We found that T6SS expression is elevated in P. aeruginosa isolated from chronically infected patients. Deletion of the retS gene activates P. aeruginosa PAO1 T6SS while repressing T3SS in vitro. Bacterial and cellular transcriptome sequencing analyses showed that T6SS genes were upregulated, while T3SS genes were downregulated in the ΔretS mutant. Additionally, the expression levels of the fimbriae gene cupC, the histidine phosphotransfer protein hptC (PA0033), and the transcription factor PA0034 were significantly increased. Subsequent experiments revealed that adhesion mediated by cupC enhances the contact-killing activity of the T6SS. Deletion of the hptC-PA0034 operon results in the down-regulation of cupC expression. The ΔretSΔcupC and ΔretSΔhptC-PA0034 mutants exhibited reduced cytotoxicity compared to the ΔretS mutant, similar to the ΔretSΔclpV1ΔclpV2 mutant. The ΔretS infection increased cell death, inflammatory factors (IL-1β, IL-6, TNF-α), and reactive oxygen species compared to a T6SS-inactive strain. Importantly, our study demonstrates that the T6SS activates the PDE4C pathway in epithelial cells, leading to significant cellular alterations. The application of PDE inhibitors effectively mitigates cell damage and inflammatory responses. These findings highlight the critical role of T6SS in modulating host cell signaling and suggest potential therapeutic strategies for conditions associated with T6SS-mediated inflammation. |
| format | Article |
| id | doaj-art-b14ea88183594a8f85afeae0cc84688e |
| institution | Kabale University |
| issn | 2235-2988 |
| language | English |
| publishDate | 2025-06-01 |
| publisher | Frontiers Media S.A. |
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| series | Frontiers in Cellular and Infection Microbiology |
| spelling | doaj-art-b14ea88183594a8f85afeae0cc84688e2025-08-20T03:45:57ZengFrontiers Media S.A.Frontiers in Cellular and Infection Microbiology2235-29882025-06-011510.3389/fcimb.2025.15823391582339Transcriptomic profiling reveals RetS-mediated regulation of type VI secretion system and host cell responses in Pseudomonas aeruginosa infectionsYinglin Wu0Yinglin Wu1Yinglin Wu2Shan Huang3Shan Huang4Shan Huang5Kai Zhang6Yangcheng Shen7Yangcheng Shen8Yangcheng Shen9Shebin Zhang10Shebin Zhang11Shebin Zhang12Haining Xia13Haining Xia14Haining Xia15Jieying Pu16Jieying Pu17Jieying Pu18Cong Shen19Cong Shen20Cong Shen21Cha Chen22Cha Chen23Cha Chen24Jianming Zeng25Jianming Zeng26Jianming Zeng27The Second Clinical Medical College, Guangzhou University of Chinese Medicine, State Key Laboratory of Traditional Chinese Medicine Syndrome, Guangdong Provincial Hospital of Chinese Medicine, Guangzhou, ChinaDepartment of Laboratory Medicine, The Second Affiliated Hospital of Guangzhou University of Chinese Medicine, Guangzhou, ChinaGuangdong Provincial Key Laboratory of Research on Emergency in Traditional Chinese Medicine (TCM), The Second Affiliated Hospital of Guangzhou University of Chinese Medicine, Guangzhou, ChinaThe Second Clinical Medical College, Guangzhou University of Chinese Medicine, State Key Laboratory of Traditional Chinese Medicine Syndrome, Guangdong Provincial Hospital of Chinese Medicine, Guangzhou, ChinaDepartment of Laboratory Medicine, The Second Affiliated Hospital of Guangzhou University of Chinese Medicine, Guangzhou, ChinaGuangdong Provincial Key Laboratory of Research on Emergency in Traditional Chinese Medicine (TCM), The Second Affiliated Hospital of Guangzhou University of Chinese Medicine, Guangzhou, ChinaSchool of Laboratory Medicine, Guangzhou Health Science College, Guangzhou, ChinaThe Second Clinical Medical College, Guangzhou University of Chinese Medicine, State Key Laboratory of Traditional Chinese Medicine Syndrome, Guangdong Provincial Hospital of Chinese Medicine, Guangzhou, ChinaDepartment of Laboratory Medicine, The Second Affiliated Hospital of Guangzhou University of Chinese Medicine, Guangzhou, ChinaGuangdong Provincial Key Laboratory of Research on Emergency in Traditional Chinese Medicine (TCM), The Second Affiliated Hospital of Guangzhou University of Chinese Medicine, Guangzhou, ChinaThe Second Clinical Medical College, Guangzhou University of Chinese Medicine, State Key Laboratory of Traditional Chinese Medicine Syndrome, Guangdong Provincial Hospital of Chinese Medicine, Guangzhou, ChinaDepartment of Laboratory Medicine, The Second Affiliated Hospital of Guangzhou University of Chinese Medicine, Guangzhou, ChinaGuangdong Provincial Key Laboratory of Research on Emergency in Traditional Chinese Medicine (TCM), The Second Affiliated Hospital of Guangzhou University of Chinese Medicine, Guangzhou, ChinaThe Second Clinical Medical College, Guangzhou University of Chinese Medicine, State Key Laboratory of Traditional Chinese Medicine Syndrome, Guangdong Provincial Hospital of Chinese Medicine, Guangzhou, ChinaDepartment of Laboratory Medicine, The Second Affiliated Hospital of Guangzhou University of Chinese Medicine, Guangzhou, ChinaGuangdong Provincial Key Laboratory of Research on Emergency in Traditional Chinese Medicine (TCM), The Second Affiliated Hospital of Guangzhou University of Chinese Medicine, Guangzhou, ChinaThe Second Clinical Medical College, Guangzhou University of Chinese Medicine, State Key Laboratory of Traditional Chinese Medicine Syndrome, Guangdong Provincial Hospital of Chinese Medicine, Guangzhou, ChinaDepartment of Laboratory Medicine, The Second Affiliated Hospital of Guangzhou University of Chinese Medicine, Guangzhou, ChinaGuangdong Provincial Key Laboratory of Research on Emergency in Traditional Chinese Medicine (TCM), The Second Affiliated Hospital of Guangzhou University of Chinese Medicine, Guangzhou, ChinaThe Second Clinical Medical College, Guangzhou University of Chinese Medicine, State Key Laboratory of Traditional Chinese Medicine Syndrome, Guangdong Provincial Hospital of Chinese Medicine, Guangzhou, ChinaDepartment of Laboratory Medicine, The Second Affiliated Hospital of Guangzhou University of Chinese Medicine, Guangzhou, ChinaGuangdong Provincial Key Laboratory of Research on Emergency in Traditional Chinese Medicine (TCM), The Second Affiliated Hospital of Guangzhou University of Chinese Medicine, Guangzhou, ChinaThe Second Clinical Medical College, Guangzhou University of Chinese Medicine, State Key Laboratory of Traditional Chinese Medicine Syndrome, Guangdong Provincial Hospital of Chinese Medicine, Guangzhou, ChinaDepartment of Laboratory Medicine, The Second Affiliated Hospital of Guangzhou University of Chinese Medicine, Guangzhou, ChinaGuangdong Provincial Key Laboratory of Research on Emergency in Traditional Chinese Medicine (TCM), The Second Affiliated Hospital of Guangzhou University of Chinese Medicine, Guangzhou, ChinaThe Second Clinical Medical College, Guangzhou University of Chinese Medicine, State Key Laboratory of Traditional Chinese Medicine Syndrome, Guangdong Provincial Hospital of Chinese Medicine, Guangzhou, ChinaDepartment of Laboratory Medicine, The Second Affiliated Hospital of Guangzhou University of Chinese Medicine, Guangzhou, ChinaGuangdong Provincial Key Laboratory of Research on Emergency in Traditional Chinese Medicine (TCM), The Second Affiliated Hospital of Guangzhou University of Chinese Medicine, Guangzhou, ChinaPseudomonas aeruginosa is a major opportunistic pathogen that causes chronic infections, particularly in patients with cystic fibrosis and chronic obstructive pulmonary disease (COPD). The type VI secretion system (T6SS) is a primary virulence factor of P. aeruginosa in chronic infections. The objective of this study was to elucidate the regulatory mechanisms and pathogenic effects of the T6SS during P. aeruginosa infection, utilizing transcriptome sequencing and functional assays. We found that T6SS expression is elevated in P. aeruginosa isolated from chronically infected patients. Deletion of the retS gene activates P. aeruginosa PAO1 T6SS while repressing T3SS in vitro. Bacterial and cellular transcriptome sequencing analyses showed that T6SS genes were upregulated, while T3SS genes were downregulated in the ΔretS mutant. Additionally, the expression levels of the fimbriae gene cupC, the histidine phosphotransfer protein hptC (PA0033), and the transcription factor PA0034 were significantly increased. Subsequent experiments revealed that adhesion mediated by cupC enhances the contact-killing activity of the T6SS. Deletion of the hptC-PA0034 operon results in the down-regulation of cupC expression. The ΔretSΔcupC and ΔretSΔhptC-PA0034 mutants exhibited reduced cytotoxicity compared to the ΔretS mutant, similar to the ΔretSΔclpV1ΔclpV2 mutant. The ΔretS infection increased cell death, inflammatory factors (IL-1β, IL-6, TNF-α), and reactive oxygen species compared to a T6SS-inactive strain. Importantly, our study demonstrates that the T6SS activates the PDE4C pathway in epithelial cells, leading to significant cellular alterations. The application of PDE inhibitors effectively mitigates cell damage and inflammatory responses. These findings highlight the critical role of T6SS in modulating host cell signaling and suggest potential therapeutic strategies for conditions associated with T6SS-mediated inflammation.https://www.frontiersin.org/articles/10.3389/fcimb.2025.1582339/fullPseudomonas aeruginosavirulencetype VI secretion systemchaperone-usher pathwayCupC fimbriaeA549 epithelial cells |
| spellingShingle | Yinglin Wu Yinglin Wu Yinglin Wu Shan Huang Shan Huang Shan Huang Kai Zhang Yangcheng Shen Yangcheng Shen Yangcheng Shen Shebin Zhang Shebin Zhang Shebin Zhang Haining Xia Haining Xia Haining Xia Jieying Pu Jieying Pu Jieying Pu Cong Shen Cong Shen Cong Shen Cha Chen Cha Chen Cha Chen Jianming Zeng Jianming Zeng Jianming Zeng Transcriptomic profiling reveals RetS-mediated regulation of type VI secretion system and host cell responses in Pseudomonas aeruginosa infections Frontiers in Cellular and Infection Microbiology Pseudomonas aeruginosa virulence type VI secretion system chaperone-usher pathway CupC fimbriae A549 epithelial cells |
| title | Transcriptomic profiling reveals RetS-mediated regulation of type VI secretion system and host cell responses in Pseudomonas aeruginosa infections |
| title_full | Transcriptomic profiling reveals RetS-mediated regulation of type VI secretion system and host cell responses in Pseudomonas aeruginosa infections |
| title_fullStr | Transcriptomic profiling reveals RetS-mediated regulation of type VI secretion system and host cell responses in Pseudomonas aeruginosa infections |
| title_full_unstemmed | Transcriptomic profiling reveals RetS-mediated regulation of type VI secretion system and host cell responses in Pseudomonas aeruginosa infections |
| title_short | Transcriptomic profiling reveals RetS-mediated regulation of type VI secretion system and host cell responses in Pseudomonas aeruginosa infections |
| title_sort | transcriptomic profiling reveals rets mediated regulation of type vi secretion system and host cell responses in pseudomonas aeruginosa infections |
| topic | Pseudomonas aeruginosa virulence type VI secretion system chaperone-usher pathway CupC fimbriae A549 epithelial cells |
| url | https://www.frontiersin.org/articles/10.3389/fcimb.2025.1582339/full |
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