Cyclosporine modulates neutrophil functions via the SIRT6–HIF‐1α–glycolysis axis to alleviate severe ulcerative colitis
Abstract Background Cyclosporine A (CsA) is routinely used to treat patients with steroid‐refractory acute severe ulcerative colitis (ASUC). Here, we studied the underlying mechanisms of CsA‐mediated alleviation in ASUC patients. Methods Neutrophil functions including expression of cytokines, apopto...
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| Format: | Article |
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Wiley
2021-02-01
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| Series: | Clinical and Translational Medicine |
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| Online Access: | https://doi.org/10.1002/ctm2.334 |
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| author | Huiying Lu Jian Lin Chunjin Xu Mingming Sun Keqiang Zuo Xiaoping Zhang Mingsong Li Hailiang Huang Zhong Li Wei Wu Baisui Feng Zhanju Liu |
| author_facet | Huiying Lu Jian Lin Chunjin Xu Mingming Sun Keqiang Zuo Xiaoping Zhang Mingsong Li Hailiang Huang Zhong Li Wei Wu Baisui Feng Zhanju Liu |
| author_sort | Huiying Lu |
| collection | DOAJ |
| description | Abstract Background Cyclosporine A (CsA) is routinely used to treat patients with steroid‐refractory acute severe ulcerative colitis (ASUC). Here, we studied the underlying mechanisms of CsA‐mediated alleviation in ASUC patients. Methods Neutrophil functions including expression of cytokines, apoptosis, and migration were measured by qRT‐PCR, flow cytometry, and Transwell assay. Dynamic changes of glycolysis and tricarboxylic acid (TCA) cycle were measured by a Seahorse extracellular flux analyzer. Gene differences were determined and verified by RNA sequencing, qRT‐PCR, and Western blotting. Small interfering RNA and inhibitors were used to knock down Sirtuin 6 (SIRT6) in HL‐60 cells and block expression of SIRT6, hypoxia‐inducible factor‐1α (HIF‐1α), and pyruvate dehydrogenase lipoamide kinase isozyme 4 (PDK4) in neutrophils. Results We found that HIF‐1α expression and glycolysis significantly increased, while the release of IL‐8, myeloperoxidase (MPO) and reactive oxygen species (ROS), the apoptosis, and ability of migration markedly decreased in neutrophils of ASUC patients who responded to CsA (Response group) compared with those who did not respond to CsA (Nonresponse group). We also observed that CsA‐induced functional alternation of neutrophils was initiated through suppressing SIRT6 expression, which is responsible for expression of the downstream signaling molecules (e.g., HIF‐1α, PFKFB3) and PDK4 ubiquitination, leading to fueling neutrophil glycolysis and TCA cycle. Furthermore, blockage of SIRT6 signaling demonstrated to be the same functional changes as CsA to decrease the migration of neutrophils. Conclusions The data reveal a novel mechanism of CsA in alleviating ASUC by promoting neutrophil HIF‐1α expression and restricting excessive neutrophil activation in a SIRT6−HIF‐1α−glycolysis axis, suggesting SIRT6 as a candidate target for maintaining mucosal homeostasis and treating intestinal inflammation. |
| first_indexed | 2024-04-10T07:08:38Z |
| format | Article |
| id | doaj.art-a34473255b2546e08c354254f260e50a |
| institution | Directory Open Access Journal |
| issn | 2001-1326 |
| language | English |
| last_indexed | 2024-04-10T07:08:38Z |
| publishDate | 2021-02-01 |
| publisher | Wiley |
| record_format | Article |
| series | Clinical and Translational Medicine |
| spelling | doaj.art-a34473255b2546e08c354254f260e50a2023-02-27T05:10:47ZengWileyClinical and Translational Medicine2001-13262021-02-01112n/an/a10.1002/ctm2.334Cyclosporine modulates neutrophil functions via the SIRT6–HIF‐1α–glycolysis axis to alleviate severe ulcerative colitisHuiying Lu0Jian Lin1Chunjin Xu2Mingming Sun3Keqiang Zuo4Xiaoping Zhang5Mingsong Li6Hailiang Huang7Zhong Li8Wei Wu9Baisui Feng10Zhanju Liu11Center for IBD Research Department of Gastroenterology Shanghai Tenth People's Hospital of Tongji University Shanghai ChinaCenter for IBD Research Department of Gastroenterology Shanghai Tenth People's Hospital of Tongji University Shanghai ChinaDepartment of Gastroenterology First People's Hospital of Shangqiu City Affiliated to Xinxiang Medical University Shangqiu ChinaCenter for IBD Research Department of Gastroenterology Shanghai Tenth People's Hospital of Tongji University Shanghai ChinaCenter for IBD Research Department of Gastroenterology Shanghai Tenth People's Hospital of Tongji University Shanghai ChinaCenter for IBD Research Department of Gastroenterology Shanghai Tenth People's Hospital of Tongji University Shanghai ChinaDepartment of Gastroenterology Third Affiliated Hospital of Guangzhou Medical University Guangzhou ChinaAnalytic and Translational Genetics Unit Massachusetts General Hospital Boston Massachusetts USAShanghai Cell Therapy Group Shanghai ChinaCenter for IBD Research Department of Gastroenterology Shanghai Tenth People's Hospital of Tongji University Shanghai ChinaDepartment of Gastroenterology Second Affiliated Hospital of Zhengzhou University Zhengzhou ChinaCenter for IBD Research Department of Gastroenterology Shanghai Tenth People's Hospital of Tongji University Shanghai ChinaAbstract Background Cyclosporine A (CsA) is routinely used to treat patients with steroid‐refractory acute severe ulcerative colitis (ASUC). Here, we studied the underlying mechanisms of CsA‐mediated alleviation in ASUC patients. Methods Neutrophil functions including expression of cytokines, apoptosis, and migration were measured by qRT‐PCR, flow cytometry, and Transwell assay. Dynamic changes of glycolysis and tricarboxylic acid (TCA) cycle were measured by a Seahorse extracellular flux analyzer. Gene differences were determined and verified by RNA sequencing, qRT‐PCR, and Western blotting. Small interfering RNA and inhibitors were used to knock down Sirtuin 6 (SIRT6) in HL‐60 cells and block expression of SIRT6, hypoxia‐inducible factor‐1α (HIF‐1α), and pyruvate dehydrogenase lipoamide kinase isozyme 4 (PDK4) in neutrophils. Results We found that HIF‐1α expression and glycolysis significantly increased, while the release of IL‐8, myeloperoxidase (MPO) and reactive oxygen species (ROS), the apoptosis, and ability of migration markedly decreased in neutrophils of ASUC patients who responded to CsA (Response group) compared with those who did not respond to CsA (Nonresponse group). We also observed that CsA‐induced functional alternation of neutrophils was initiated through suppressing SIRT6 expression, which is responsible for expression of the downstream signaling molecules (e.g., HIF‐1α, PFKFB3) and PDK4 ubiquitination, leading to fueling neutrophil glycolysis and TCA cycle. Furthermore, blockage of SIRT6 signaling demonstrated to be the same functional changes as CsA to decrease the migration of neutrophils. Conclusions The data reveal a novel mechanism of CsA in alleviating ASUC by promoting neutrophil HIF‐1α expression and restricting excessive neutrophil activation in a SIRT6−HIF‐1α−glycolysis axis, suggesting SIRT6 as a candidate target for maintaining mucosal homeostasis and treating intestinal inflammation.https://doi.org/10.1002/ctm2.334glycolysisneutrophilSIRT6ulcerative colitis |
| spellingShingle | Huiying Lu Jian Lin Chunjin Xu Mingming Sun Keqiang Zuo Xiaoping Zhang Mingsong Li Hailiang Huang Zhong Li Wei Wu Baisui Feng Zhanju Liu Cyclosporine modulates neutrophil functions via the SIRT6–HIF‐1α–glycolysis axis to alleviate severe ulcerative colitis Clinical and Translational Medicine glycolysis neutrophil SIRT6 ulcerative colitis |
| title | Cyclosporine modulates neutrophil functions via the SIRT6–HIF‐1α–glycolysis axis to alleviate severe ulcerative colitis |
| title_full | Cyclosporine modulates neutrophil functions via the SIRT6–HIF‐1α–glycolysis axis to alleviate severe ulcerative colitis |
| title_fullStr | Cyclosporine modulates neutrophil functions via the SIRT6–HIF‐1α–glycolysis axis to alleviate severe ulcerative colitis |
| title_full_unstemmed | Cyclosporine modulates neutrophil functions via the SIRT6–HIF‐1α–glycolysis axis to alleviate severe ulcerative colitis |
| title_short | Cyclosporine modulates neutrophil functions via the SIRT6–HIF‐1α–glycolysis axis to alleviate severe ulcerative colitis |
| title_sort | cyclosporine modulates neutrophil functions via the sirt6 hif 1α glycolysis axis to alleviate severe ulcerative colitis |
| topic | glycolysis neutrophil SIRT6 ulcerative colitis |
| url | https://doi.org/10.1002/ctm2.334 |
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