Cysteine thiol oxidation on SIRT2 regulates inflammation in obese mice with sepsis
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Obesity increases morbidity and mortality in acute illnesses such as sepsis and septic shock. We showed previously that the early/hyper-inflammatory phase of sepsis is exaggerated in obese mice with sepsis; sirtuin 2 (SIRT2) modulates sepsis inflammation in obesity. Evidence suggests that obesity with sepsis is associated with increased oxidative stress. It is unknown whether exaggerated hyper-inflammation of obesity with sepsis modulates the SIRT2 function in return. We showed recently that SIRT6 oxidation during hyper-inflammation of sepsis modulates its glycolytic function. This study tested the hypothesis that increased oxidative stress and direct SIRT2 oxidation exaggerate hyper-inflammation in obesity with sepsis. Using spleen and liver tissue from mice with diet-induced obesity (DIO) we studied oxidized vs. total SIRT2 expression during hyper- and hypo-inflammation of sepsis. To elucidate the mechanism of SIRT2 oxidation (specific modifications of redox-sensitive cysteines) and its effect on inflammation, we performed site-directed mutations of redox-sensitive cysteines Cys221 and Cys224 on SIRT2 to serine (C221S and C224S), transfected HEK293 cells with mutants or WT SIRT2, and studied SIRT2 enzymatic activity and NFĸBp65 deacetylation. Finally, we studied the effect of SIRT2 mutation on LPS-induced inflammation using RAW 264.7 macrophages. In an inverse relationship, total SIRT2 decreased while oxidized SIRT2 expression increased during hyper-inflammation and SIRT2 was unable to deacetylate NFĸBp65 with increased oxidative stress of obesity with sepsis. Mechanistically, both the mutants (C221S and C224S) show decreased (1) SIRT2 enzymatic activity, (2) deacetylation of NFĸBp65, and (3) anti-inflammatory activity in response to LPS vs. WT SIRT2. Direct oxidation modulates SIRT2 function during hyper-inflammatory phase of obesity with sepsis via redox sensitive cysteines.
KEY WORDSobesity sepsis septic shock hyper-inflammation oxidative stress
Cecal ligation and puncture
Control diet mice
Diet induced obesity
Nuclear factor kappa B
Wild type (lean)
The plasmids were gifted to us by Addgene; pcDNA3β-FLAG-CBP-HA was a gift from Tso-Pang Yao (Addgene plasmid #32908); pCMV4 NFĸB p65 was a gift from Warner Greene (Addgene plasmid #21966). Wild-type plasmid SIRT2 flag was a gift from Eric Verdin (Addgene plasmid #13813).
Concept and design: VTV and CF; data collection: XW, NB, DL and VTV; data analysis and interpretation: VTV, XW, CEM and CF; generating the manuscript: VTV, XW, CEM and CF.
This work was supported by NIH grants: Vidula T. Vachharajani, R01GM099807; Charles E McCall, (1) R01AI065791, (2) R01AI079144 (3) 1R35GM126922.
Compliance with Ethical Standards
Conflict of Interest
The authors declare that the submitted work was not carried out in the presence of any personal, professional or financial relationships that could potentially be construed as a conflict of interest.
- 2.Torio, C. M., and B. J. Moore. 2006. National Inpatient Hospital Costs: The Most Expensive Conditions by Payer, 2013: Statistical Brief #204. In Healthcare Cost and Utilization Project (HCUP) Statistical Briefs. Rockville (MD).Google Scholar
- 5.Vachharajani, V.T., T. Fu Liu, C.M. Brown, X. Wang, N.L. Buechler, J.D. Wells, B.K. Yoza, and C.E. McCall. 2014. SIRT1 inhibition during the hypoinflammatory phenotype of sepsis enhances immunity and improves outcome. Journal of Leukocyte Biology. https://doi.org/10.1189/jlb.3MA0114-034RR.
- 8.Goncalves Damascena, K., C. Batisti Ferreira, P. Dos Santos Teixeira, B. Madrid, A. Goncalves, C. Cordova, O. de Toledo Nobrega, and A. Pimentel Ferreira. 2017. Functional capacity and obesity reflect the cognitive performance of older adults living in long-term care facilities. Psychogeriatrics 17 (6): 439–445. https://doi.org/10.1111/psyg.12273.CrossRefPubMedGoogle Scholar
- 9.Oguri, M., T. Fujimaki, H. Horibe, K. Kato, K. Matsui, I. Takeuchi, and Y. Yamada. 2017. Obesity-related changes in clinical parameters and conditions in a longitudinal population-based epidemiological study. Obes Res Clin Pract 11 (3): 299–314. https://doi.org/10.1016/j.orcp.2016.08.008.CrossRefPubMedGoogle Scholar
- 11.Arabi, Y. M., S. I. Dara, H. M. Tamim, A. H. Rishu, A. Bouchama, M. K. Khedr, D. Feinstein et al. . 2013. Clinical characteristics, sepsis interventions and outcomes in the obese patients with septic shock: An international multicenter cohort study. Critical Care 17 (2):R72. https://doi.org/10.1186/cc12680.
- 12.Prescott, H.C., V.W. Chang, J.M. O'Brien Jr., K.M. Langa, and T. Iwashyna. 2014. Obesity and 1-year outcomes in older Americans with severe sepsis. Critical Care Medicine. https://doi.org/10.1097/CCM.0000000000000336.
- 13.Robinson, M.K., K.M. Mogensen, J.D. Casey, C.K. McKane, T. Moromizato, J.D. Rawn, and K.B. Christopher. 2015. The relationship among obesity, nutritional status, and mortality in the critically ill. Critical Care Medicine 43 (1): 87–100. https://doi.org/10.1097/CCM.0000000000000602.CrossRefPubMedGoogle Scholar
- 17.Vachharajani, V., S. Vital, and J. Russell. 2010. Modulation of circulating cell-endothelial cell interaction by erythropoietin in lean and obese mice with cecal ligation and puncture. Pathophysiology : the official journal of the International Society for Pathophysiology / ISP 17 (1): 9–18. https://doi.org/10.1016/j.pathophys.2009.04.002. CrossRefGoogle Scholar
- 20.Wang, X., N.L. Buechler, B.K. Yoza, C.E. McCall, and V.T. Vachharajani. 2015. Resveratrol attenuates microvascular inflammation in sepsis via SIRT-1-induced modulation of adhesion molecules in ob/ob mice. Obesity (Silver Spring) 23 (6): 1209–1217. https://doi.org/10.1002/oby.21086.CrossRefGoogle Scholar
- 21.Liu, T.F., V. Vachharajani, P. Millet, M.S. Bharadwaj, A.J. Molina, and C.E. McCall. 2015. Sequential actions of SIRT1-RELB-SIRT3 coordinate nuclear-mitochondrial communication during immunometabolic adaptation to acute inflammation and sepsis. The Journal of Biological Chemistry 290 (1): 396–408. https://doi.org/10.1074/jbc.M114.566349.CrossRefPubMedGoogle Scholar
- 22.Liu, T.F., V.T. Vachharajani, B.K. Yoza, and C.E. McCall. 2012. NAD+−dependent sirtuin 1 and 6 proteins coordinate a switch from glucose to fatty acid oxidation during the acute inflammatory response. The Journal of Biological Chemistry 287 (31): 25758–25769. https://doi.org/10.1074/jbc.M112.362343.CrossRefPubMedPubMedCentralGoogle Scholar
- 23.Liu, T.F., B.K. Yoza, M. El Gazzar, V.T. Vachharajani, and C.E. McCall. 2011. NAD+−dependent SIRT1 deacetylase participates in epigenetic reprogramming during endotoxin tolerance. The Journal of Biological Chemistry 286 (11): 9856–9864. https://doi.org/10.1074/jbc.M110.196790.CrossRefPubMedPubMedCentralGoogle Scholar
- 24.Kauppinen, A., T. Suuronen, J. Ojala, K. Kaarniranta, and A. Salminen. 2013. Antagonistic crosstalk between NF-kappaB and SIRT1 in the regulation of inflammation and metabolic disorders. Cellular Signalling 25 (10): 1939–1948. https://doi.org/10.1016/j.cellsig.2013.06.007.CrossRefPubMedGoogle Scholar
- 27.Haigis, M.C., and D.A. Sinclair. 2010. Mammalian sirtuins: Biological insights and disease relevance. Annual Review of Pathology 5: 253–295. https://doi.org/10.1146/annurev.pathol.4.110807.092250.CrossRefPubMedPubMedCentralGoogle Scholar
- 30.Mariani, S., G. Di Rocco, G. Toietta, M.A. Russo, E. Petrangeli, and L. Salvatori. 2017. Sirtuins 1-7 expression in human adipose-derived stem cells from subcutaneous and visceral fat depots: Influence of obesity and hypoxia. Endocrine 57 (3): 455–463. https://doi.org/10.1007/s12020-016-1170-8. CrossRefPubMedGoogle Scholar
- 31.Moschen, A.R., V. Wieser, R.R. Gerner, A. Bichler, B. Enrich, P. Moser, C.F. Ebenbichler, S. Kaser, and H. Tilg. 2013. Adipose tissue and liver expression of SIRT1, 3, and 6 increase after extensive weight loss in morbid obesity. Journal of Hepatology 59 (6): 1315–1322. https://doi.org/10.1016/j.jhep.2013.07.027.CrossRefPubMedGoogle Scholar
- 32.Krishnan, J., C. Danzer, T. Simka, J. Ukropec, K.M. Walter, S. Kumpf, P. Mirtschink, et al. 2012. Dietary obesity-associated Hif1alpha activation in adipocytes restricts fatty acid oxidation and energy expenditure via suppression of the Sirt2-NAD+ system. Genes & Development 26 (3): 259–270. https://doi.org/10.1101/gad.180406.111. CrossRefGoogle Scholar
- 34.Haden, D.W., H.B. Suliman, M.S. Carraway, K.E. Welty-Wolf, A.S. Ali, H. Shitara, H. Yonekawa, and C.A. Piantadosi. 2007. Mitochondrial biogenesis restores oxidative metabolism during Staphylococcus aureus sepsis. American Journal of Respiratory and Critical Care Medicine 176 (8): 768–777. https://doi.org/10.1164/rccm.200701-161OC.CrossRefPubMedPubMedCentralGoogle Scholar
- 36.Jung, S.B., C.S. Kim, Y.R. Kim, A. Naqvi, T. Yamamori, S. Kumar, A. Kumar, and K. Irani. 2013. Redox factor-1 activates endothelial SIRTUIN1 through reduction of conserved cysteine sulfhydryls in its deacetylase domain. PLoS One 8 (6): e65415. https://doi.org/10.1371/journal.pone.0065415.CrossRefPubMedPubMedCentralGoogle Scholar
- 37.Chen, L., Y. Feng, Y. Zhou, W. Zhu, X. Shen, K. Chen, H. Jiang, and D. Liu. 2010. Dual role of Zn2+ in maintaining structural integrity and suppressing deacetylase activity of SIRT1. Journal of Inorganic Biochemistry 104 (2): 180–185. https://doi.org/10.1016/j.jinorgbio.2009.10.021.CrossRefPubMedGoogle Scholar
- 39.Long, D., H. Wu, A.W. Tsang, L.B. Poole, B.K. Yoza, X. Wang, V. Vachharajani, C.M. Furdui, and C.E. McCall. 2017. The oxidative state of cysteine thiol 144 regulates the SIRT6 glucose homeostat. Scientific Reports 7 (1): 11005. https://doi.org/10.1038/s41598-017-11388-6.CrossRefPubMedPubMedCentralGoogle Scholar
- 40.Zee, R.S., C.B. Yoo, D.R. Pimentel, D.H. Perlman, J.R. Burgoyne, X. Hou, M.E. McComb, C.E. Costello, R.A. Cohen, and M.M. Bachschmid. 2010. Redox regulation of sirtuin-1 by S-glutathiolation. Antioxidants & Redox Signaling 13 (7): 1023–1032. https://doi.org/10.1089/ars.2010.3251.CrossRefGoogle Scholar
- 42.Furukawa, S., T. Fujita, M. Shimabukuro, M. Iwaki, Y. Yamada, Y. Nakajima, O. Nakayama, M. Makishima, M. Matsuda, and I. Shimomura. 2004. Increased oxidative stress in obesity and its impact on metabolic syndrome. The Journal of Clinical Investigation 114 (12): 1752–1761. https://doi.org/10.1172/JCI21625.CrossRefPubMedPubMedCentralGoogle Scholar
- 45.Vachharajani, V.T., T. Liu, C.M. Brown, X. Wang, N.L. Buechler, J.D. Wells, B.K. Yoza, and C.E. McCall. 2014. SIRT1 inhibition during the hypoinflammatory phenotype of sepsis enhances immunity and improves outcome. Journal of Leukocyte Biology 96 (5): 785–796. https://doi.org/10.1189/jlb.3MA0114-034RR.CrossRefPubMedPubMedCentralGoogle Scholar
- 47.Zhao, X., T. Sternsdorf, T.A. Bolger, R.M. Evans, and T.P. Yao. 2005. Regulation of MEF2 by histone deacetylase 4- and SIRT1 deacetylase-mediated lysine modifications. Molecular and Cellular Biology 25 (19): 8456–8464. https://doi.org/10.1128/MCB.25.19.8456-8464.2005.CrossRefPubMedPubMedCentralGoogle Scholar
- 48.Ballard, D.W., E.P. Dixon, N.J. Peffer, H. Bogerd, S. Doerre, B. Stein, and W.C. Greene. 1992. The 65-kDa subunit of human NF-kappa B functions as a potent transcriptional activator and a target for v-Rel-mediated repression. Proceedings of the National Academy of Sciences of the United States of America 89 (5): 1875–1879.CrossRefPubMedPubMedCentralGoogle Scholar
- 53.Jung, U., K.E. Norman, K. Scharffetter-Kochanek, A.L. Beaudet, and K. Ley. 1998. Transit time of leukocytes rolling through venules controls cytokine-induced inflammatory cell recruitment in vivo. The Journal of Clinical Investigation 102 (8): 1526–1533. https://doi.org/10.1172/JCI119893.CrossRefPubMedPubMedCentralGoogle Scholar
- 58.Chang, J., S.L. Kunkel, and C.H. Chang. 2009. Negative regulation of MyD88-dependent signaling by IL-10 in dendritic cells. Proceedings of the National Academy of Sciences of the United States of America 106 (43): 18327–18332. https://doi.org/10.1073/pnas.0905815106.CrossRefPubMedPubMedCentralGoogle Scholar
- 59.Li, Y. P., J. Huang, S. G. Huang, Y. G. Xu, Y. Y. Xu, J. Y. Liao, X. Feng, X. G. Zhang, J. H. Wang, and J. Wang. 2014. The compromised inflammatory response to bacterial components after pediatric cardiac surgery is associated with cardiopulmonary bypass-suppressed toll-like receptor signal transduction pathways. Journal of Critical Care 29 (2):312 e317–313. https://doi.org/10.1016/j.jcrc.2013.10.008.
- 60.Eo, S.H., S.Y. Choi, and S.J. Kim. 2016. PEP-1-SIRT2-induced matrix metalloproteinase-1 and -13 modulates type II collagen expression via ERK signaling in rabbit articular chondrocytes. Experimental Cell Research 348 (2): 201–208. https://doi.org/10.1016/j.yexcr.2016.09.024.CrossRefPubMedGoogle Scholar
- 63.Wu, D., W. Lu, Z. Wei, M. Xu, and X. Liu. 2018. Neuroprotective effect of Sirt2-specific inhibitor AK-7 against acute cerebral ischemia is P38 activation-dependent in mice. Neuroscience 374: 61–69. https://doi.org/10.1016/j.neuroscience.2018.01.040.CrossRefPubMedGoogle Scholar
- 67.Pandithage, R., R. Lilischkis, K. Harting, A. Wolf, B. Jedamzik, J. Luscher-Firzlaff, J. Vervoorts, et al. 2008. The regulation of SIRT2 function by cyclin-dependent kinases affects cell motility. The Journal of Cell Biology 180 (5): 915–929. https://doi.org/10.1083/jcb.200707126. CrossRefPubMedPubMedCentralGoogle Scholar
- 68.Ramakrishnan, G., G. Davaakhuu, L. Kaplun, W.C. Chung, A. Rana, A. Atfi, L. Miele, and G. Tzivion. 2014. Sirt2 deacetylase is a novel AKT binding partner critical for AKT activation by insulin. The Journal of Biological Chemistry 289 (9): 6054–6066. https://doi.org/10.1074/jbc.M113.537266.CrossRefPubMedPubMedCentralGoogle Scholar
- 69.Hu, S., H. Liu, Y. Ha, X. Luo, M. Motamedi, M.P. Gupta, J.X. Ma, R.G. Tilton, and W. Zhang. 2015. Posttranslational modification of Sirt6 activity by peroxynitrite. Free Radical Biology & Medicine 79: 176–185. https://doi.org/10.1016/j.freeradbiomed.2014.11.011.CrossRefGoogle Scholar