{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,18]],"date-time":"2026-09-18T11:31:53Z","timestamp":1789731113583,"version":"4.0.1"},"reference-count":50,"publisher":"MDPI AG","issue":"7","license":[{"start":{"date-parts":[[2025,7,20]],"date-time":"2025-07-20T00:00:00Z","timestamp":1752969600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100002584","name":"Department of Anesthesiology (Duke University Medical Center)","doi-asserted-by":"publisher","award":["19IPLOI34760123"],"award-info":[{"award-number":["19IPLOI34760123"]}],"id":[{"id":"10.13039\/100002584","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100002584","name":"Department of Anesthesiology (Duke University Medical Center)","doi-asserted-by":"publisher","award":["HL130443"],"award-info":[{"award-number":["HL130443"]}],"id":[{"id":"10.13039\/100002584","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100002584","name":"Department of Anesthesiology (Duke University Medical Center)","doi-asserted-by":"publisher","award":["HL157354"],"award-info":[{"award-number":["HL157354"]}],"id":[{"id":"10.13039\/100002584","id-type":"DOI","asserted-by":"publisher"}]},{"name":"NIH","award":["19IPLOI34760123"],"award-info":[{"award-number":["19IPLOI34760123"]}]},{"name":"NIH","award":["HL130443"],"award-info":[{"award-number":["HL130443"]}]},{"name":"NIH","award":["HL157354"],"award-info":[{"award-number":["HL157354"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Pharmaceuticals"],"abstract":"<jats:p>Introduction: Accumulating evidence indicates that neutrophils undergo reprogramming of their effector functions as they migrate from the bloodstream into an inflamed tissue. Here, we examined the role of the small ubiquitin-like modifier (SUMO) conjugation in modulating neutrophil functional changes in the inflammatory microenvironment. Methods: Primary human and murine neutrophils were used to assess SUMOylation levels in vitro by Western blotting and results were validated in clinical samples from patients undergoing surgery involving hypothermic circulatory arrest. SUMOylation was inhibited with TAK-981, and its impact on neutrophil migration, NETosis, and phagocytosis was assessed in vitro. The in vivo effect of TAK-981 on neutrophil tissue infiltration was further evaluated using a sterile sponge assay in mice. Results: Our results demonstrated that neutrophil SUMOylation was induced by factors of the inflammatory microenvironment (temperature and oxidative stress) and inflammatory stimulants in vitro, and under conditions of general inflammatory activation in patients. Further, we found that blocking SUMOylation with TAK-981 in vitro blunted neutrophil migration and phagocytosis but did not affect NETosis. Notably, TAK-981 treatment reduced neutrophil accumulation in sterile sponges in mice. Conclusions: Our work identifies SUMOylation as a novel mechanism of neutrophil tissue reprogramming. Blocking SUMOylation may provide a therapeutic option to limit the contribution of neutrophils to inflammation-associated tissue damage.<\/jats:p>","DOI":"10.3390\/ph18071070","type":"journal-article","created":{"date-parts":[[2025,7,21]],"date-time":"2025-07-21T07:56:14Z","timestamp":1753084574000},"page":"1070","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["SUMOylation Regulates Neutrophil Phagocytosis and Migration"],"prefix":"10.3390","volume":"18","author":[{"given":"Ran","family":"Zhang","sequence":"first","affiliation":[{"name":"Department of Anesthesiology, Duke University Medical Center, Durham, NC 27710, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wanying","family":"Miao","sequence":"additional","affiliation":[{"name":"Department of Anesthesiology, Duke University Medical Center, Durham, NC 27710, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jin","family":"Zhang","sequence":"additional","affiliation":[{"name":"Department of Anesthesiology, Duke University Medical Center, Durham, NC 27710, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xinyuan","family":"Yu","sequence":"additional","affiliation":[{"name":"Department of Anesthesiology, Duke University Medical Center, Durham, NC 27710, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lihong","family":"Dang","sequence":"additional","affiliation":[{"name":"Department of Anesthesiology, Duke University Medical Center, Durham, NC 27710, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ata Ur","family":"Rehman","sequence":"additional","affiliation":[{"name":"Department of Anesthesiology, Duke University Medical Center, Durham, NC 27710, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5270-6353","authenticated-orcid":false,"given":"Feng","family":"Xu","sequence":"additional","affiliation":[{"name":"Department of Anesthesiology, Duke University Medical Center, Durham, NC 27710, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4325-2940","authenticated-orcid":false,"given":"Huaxin","family":"Sheng","sequence":"additional","affiliation":[{"name":"Department of Anesthesiology, Duke University Medical Center, Durham, NC 27710, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"G. Chad","family":"Hughes","sequence":"additional","affiliation":[{"name":"Department of Surgery, Division of Thoracic and Cardiovascular Surgery, Duke University Medical Center, Durham, NC 27710, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Joseph P.","family":"Mathew","sequence":"additional","affiliation":[{"name":"Department of Anesthesiology, Duke University Medical Center, Durham, NC 27710, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"J\u00f6rn","family":"Karhausen","sequence":"additional","affiliation":[{"name":"Department of Anesthesiology, Duke University Medical Center, Durham, NC 27710, USA"},{"name":"Department of Anesthesiology, IRCCS Humanitas Research Hospital, 20089 Rozzano, MI, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5719-4393","authenticated-orcid":false,"given":"Wei","family":"Yang","sequence":"additional","affiliation":[{"name":"Department of Anesthesiology, Duke University Medical Center, Durham, NC 27710, USA"},{"name":"Department of Neurology, Duke University Medical Center, Durham, NC 27710, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2025,7,20]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"1377","DOI":"10.1016\/j.immuni.2021.06.006","article-title":"The Neutrophil","volume":"54","author":"Burn","year":"2021","journal-title":"Immunity"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"565","DOI":"10.1016\/j.it.2019.04.012","article-title":"Neutrophil Diversity in Health and Disease","volume":"40","author":"Fridlender","year":"2019","journal-title":"Trends Immunol."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"575","DOI":"10.1038\/s41590-023-01452-x","article-title":"Strategies of neutrophil diversification","volume":"24","author":"Sicilia","year":"2023","journal-title":"Nat. Immunol."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"1282","DOI":"10.1016\/j.cell.2020.10.003","article-title":"Co-option of Neutrophil Fates by Tissue Environments","volume":"183","author":"Ballesteros","year":"2020","journal-title":"Cell"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"1196","DOI":"10.1182\/blood-2009-11-254490","article-title":"CCR7 is involved in the migration of neutrophils to lymph nodes","volume":"117","author":"Beauvillain","year":"2011","journal-title":"Blood"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"715","DOI":"10.1038\/s41580-022-00500-y","article-title":"Signalling mechanisms and cellular functions of SUMO","volume":"23","author":"Vertegaal","year":"2022","journal-title":"Nat. Rev. Mol. Cell Biol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"rs4","DOI":"10.1126\/scisignal.2001484","article-title":"Comparative Proteomic Analysis Identifies a Role for SUMO in Protein Quality Control","volume":"4","author":"Tatham","year":"2011","journal-title":"Sci. Signal."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"349","DOI":"10.1016\/j.molcel.2005.12.019","article-title":"Regulation of SUMOylation by Reversible Oxidation of SUMO Conjugating Enzymes","volume":"21","author":"Bossis","year":"2006","journal-title":"Mol. Cell"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"1197","DOI":"10.1074\/mcp.RA119.001401","article-title":"Hypoxia-induced Changes in SUMO Conjugation Affect Transcriptional Regulation Under Low Oxygen","volume":"18","author":"Chachami","year":"2019","journal-title":"Mol. Cell. Proteom."},{"key":"ref_10","doi-asserted-by":"crossref","unstructured":"Joshi, G., Arya, P., Mahajan, V., Chaturvedi, A., and Mishra, R.K. (2021). SUMO and SUMOylation Pathway at the Forefront of Host Immune Response. Front. Cell Dev. Biol., 9.","DOI":"10.3389\/fcell.2021.681057"},{"key":"ref_11","doi-asserted-by":"crossref","unstructured":"Karhausen, J., Ulloa, L., and Yang, W. (2021). SUMOylation Connects Cell Stress Responses and Inflammatory Control: Lessons From the Gut as a Model Organ. Front. Immunol., 12.","DOI":"10.3389\/fimmu.2021.646633"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"140","DOI":"10.1038\/ni.3342","article-title":"Sumoylation coordinates the repression of inflammatory and anti-viral gene-expression programs during innate sensing","volume":"17","author":"Decque","year":"2016","journal-title":"Nat. Immunol."},{"key":"ref_13","doi-asserted-by":"crossref","unstructured":"Zitti, B., Molfetta, R., Fionda, C., Quatrini, L., Stabile, H., Lecce, M., de Turris, V., Ricciardi, M.R., Petrucci, M.T., and Cippitelli, M. (2017). Innate immune activating ligand SUMOylation affects tumor cell recognition by NK cells. Sci. Rep., 7.","DOI":"10.1038\/s41598-017-10403-0"},{"key":"ref_14","doi-asserted-by":"crossref","unstructured":"Huang, C.-H., Yang, T.-T., and Lin, K.-I. (2024). Mechanisms and functions of SUMOylation in health and disease: A review focusing on immune cells. J. Biomed. Sci., 31.","DOI":"10.1186\/s12929-024-01003-y"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"40867","DOI":"10.1074\/jbc.M111.279984","article-title":"Mechanistic Studies of Substrate-assisted Inhibition of Ubiquitin-activating Enzyme by Adenosine Sulfamate Analogues","volume":"286","author":"Chen","year":"2011","journal-title":"J. Biol. Chem."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"2501","DOI":"10.1021\/acs.jmedchem.0c01491","article-title":"Discovery of TAK-981, a First-in-Class Inhibitor of SUMO-Activating Enzyme for the Treatment of Cancer","volume":"64","author":"Langston","year":"2021","journal-title":"J. Med. Chem."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"58","DOI":"10.1186\/s40164-023-00420-3","article-title":"The emerging roles of SUMOylation in the tumor microenvironment and therapeutic implications","volume":"12","author":"Gu","year":"2023","journal-title":"Exp. Hematol. Oncol."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"886","DOI":"10.1038\/jcbfm.2009.16","article-title":"Deep Hypothermia Markedly Activates the Small Ubiquitin-Like Modifier Conjugation Pathway; Implications for the Fate of Cells Exposed to Transient Deep Hypothermic Cardiopulmonary Bypass","volume":"29","author":"Yang","year":"2009","journal-title":"J. Cereb. Blood Flow Metab."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"68","DOI":"10.1016\/j.jtcvs.2016.05.063","article-title":"Mast cell activation and arterial hypotension during proximal aortic repair requiring hypothermic circulatory arrest","volume":"153","author":"Kertai","year":"2016","journal-title":"J. Thorac. Cardiovasc. Surg."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"349","DOI":"10.1111\/j.1471-4159.2012.07916.x","article-title":"Moderate hypothermia induces marked increase in levels and nuclear accumulation of SUMO2\/3-conjugated proteins in neurons","volume":"123","author":"Wang","year":"2012","journal-title":"J. Neurochem."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"378","DOI":"10.1038\/nri.2016.49","article-title":"Neutrophil migration in infection and wound repair: Going forward in reverse","volume":"16","author":"Rosowski","year":"2016","journal-title":"Nat. Rev. Immunol."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"135","DOI":"10.1038\/s41590-019-0571-2","article-title":"Programmed \u2018disarming\u2019 of the neutrophil proteome reduces the magnitude of inflammation","volume":"21","author":"Adrover","year":"2020","journal-title":"Nat. Immunol."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"390","DOI":"10.1016\/j.immuni.2019.01.002","article-title":"A Neutrophil Timer Coordinates Immune Defense and Vascular Protection","volume":"50","author":"Adrover","year":"2019","journal-title":"Immunity"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"eadf6493","DOI":"10.1126\/science.adf6493","article-title":"Deterministic reprogramming of neutrophils within tumors","volume":"383","author":"Ng","year":"2024","journal-title":"Science"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"528","DOI":"10.1038\/nature15367","article-title":"Neutrophil ageing is regulated by the microbiome","volume":"525","author":"Zhang","year":"2015","journal-title":"Nature"},{"key":"ref_26","doi-asserted-by":"crossref","unstructured":"D\u00f6lling, M., Eckstein, M., Singh, J., Schauer, C., Schoen, J., Shan, X., Bozec, A., Knopf, J., Schett, G., and Mu\u00f1oz, L.E. (2022). Hypoxia Promotes Neutrophil Survival After Acute Myocardial Infarction. Front. Immunol., 13.","DOI":"10.3389\/fimmu.2022.726153"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"1367","DOI":"10.1002\/JLB.4A0319-079RR","article-title":"Prolonged exposure to hypoxia induces an autophagy-like cell survival program in human neutrophils","volume":"106","author":"Talla","year":"2019","journal-title":"J. Leukoc. Biol."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"136","DOI":"10.4049\/jimmunol.2100475","article-title":"Extracellular pH Controls Chemotaxis of Neutrophil Granulocytes by Regulating Leukotriene B4 Production and Cdc42 Signaling","volume":"209","author":"Oster","year":"2022","journal-title":"J. Immunol."},{"key":"ref_29","doi-asserted-by":"crossref","unstructured":"Khachaturyan, G., Holle, A.W., Ende, K., Frey, C., Schwederski, H.A., Eiseler, T., Paschke, S., Micoulet, A., Spatz, J.P., and Kemkemer, R. (2022). Temperature-sensitive migration dynamics in neutrophil-differentiated HL-60 cells. Sci. Rep., 12.","DOI":"10.1038\/s41598-022-10858-w"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"290","DOI":"10.1007\/s00018-024-05319-8","article-title":"Unconventional posttranslational modification in innate immunity","volume":"81","author":"Chen","year":"2024","journal-title":"Cell. Mol. Life Sci."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"38","DOI":"10.1016\/j.semcdb.2021.12.010","article-title":"Stress\u2014Regulation of SUMO conjugation and of other Ubiquitin-Like Modifiers","volume":"132","author":"Ilic","year":"2022","journal-title":"Semin. Cell Dev. Biol."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"799","DOI":"10.1038\/s41374-018-0035-6","article-title":"Ubc9 overexpression and SUMO1 deficiency blunt inflammation after intestinal ischemia\/reperfusion","volume":"98","author":"Karhausen","year":"2018","journal-title":"Lab. Investig."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"965","DOI":"10.15252\/embr.201338386","article-title":"Sumoylation controls host anti-bacterial response to the gut invasive pathogen Shigella flexneri","volume":"15","author":"Fritah","year":"2014","journal-title":"Embo Rep."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"1192","DOI":"10.1038\/nature08963","article-title":"Listeria monocytogenes impairs SUMOylation for efficient infection","volume":"464","author":"Ribet","year":"2010","journal-title":"Nature"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"2266","DOI":"10.1136\/gutjnl-2021-324834","article-title":"Targeting pancreatic cancer by TAK-981: A SUMOylation inhibitor that activates the immune system and blocks cancer cell cycle progression in a preclinical model","volume":"71","author":"Kumar","year":"2022","journal-title":"Gut"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"eaba7791","DOI":"10.1126\/scitranslmed.aba7791","article-title":"A small-molecule SUMOylation inhibitor activates antitumor immune responses and potentiates immune therapies in preclinical models","volume":"13","author":"Lightcap","year":"2021","journal-title":"Sci. Transl. Med."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"878","DOI":"10.15252\/embr.201438534","article-title":"SUMO2 is essential while SUMO3 is dispensable for mouse embryonic development","volume":"15","author":"Wang","year":"2014","journal-title":"Embo Rep."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"4106","DOI":"10.1242\/jcs.038570","article-title":"Loss of SUMO1 in mice affects RanGAP1 localization and formation of PML nuclear bodies, but is not lethal as it can be compensated by SUMO2 or SUMO3","volume":"121","author":"Evdokimov","year":"2008","journal-title":"J. Cell Sci."},{"key":"ref_39","first-page":"98","article-title":"SUMOylation inhibitor TAK-981 (subasumstat) synergizes with 5-azacitidine in preclinical models of acute myeloid leukemia","volume":"109","author":"Gabellier","year":"2024","journal-title":"Haematologica"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"2149","DOI":"10.1182\/blood-2018-12-844605","article-title":"Neutrophil transendothelial migration: Updates and new perspectives","volume":"133","author":"Filippi","year":"2019","journal-title":"Blood"},{"key":"ref_41","first-page":"32","article-title":"Linking the SUMO protease SENP5 to neutrophil differentiation of AML cells","volume":"4","author":"Federzoni","year":"2015","journal-title":"Leuk. Res. Rep."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"417","DOI":"10.1016\/j.yexcr.2018.07.005","article-title":"Talin is a substrate for SUMOylation in migrating cancer cells","volume":"370","author":"Huang","year":"2018","journal-title":"Exp. Cell Res."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"871","DOI":"10.1074\/mcp.RA117.000014","article-title":"Global Identification of Small Ubiquitin-related Modifier (SUMO) Substrates Reveals Crosstalk between SUMOylation and Phosphorylation Promotes Cell Migration","volume":"17","author":"Uzoma","year":"2018","journal-title":"Mol. Cell. Proteom."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"834","DOI":"10.1038\/s41467-020-14581-w","article-title":"Quantitative SUMO proteomics identifies PIAS1 substrates involved in cell migration and motility","volume":"11","author":"Li","year":"2020","journal-title":"Nat. Commun."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"496","DOI":"10.1016\/j.trecan.2020.11.009","article-title":"Targeting SUMO Signaling to Wrestle Cancer","volume":"7","author":"Kroonen","year":"2021","journal-title":"Trends Cancer"},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"658","DOI":"10.1161\/CIRCULATIONAHA.123.067022","article-title":"Cognitive Effects of Body Temperature During Hypothermic Circulatory Arrest Trial (GOT ICE): A Randomized Clinical Trial Comparing Outcomes After Aortic Arch Surgery","volume":"149","author":"Hughes","year":"2024","journal-title":"Circulation"},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"1690","DOI":"10.1038\/srep01690","article-title":"Analysis of SUMOylated proteins using SUMO-traps","volume":"3","author":"Xolalpa","year":"2013","journal-title":"Sci. Rep."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"100948","DOI":"10.1016\/j.xpro.2021.100948","article-title":"Protocol for analysis of mouse neutrophil NETosis by flow cytometry","volume":"2","author":"McGill","year":"2021","journal-title":"STAR Protoc."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"95","DOI":"10.1186\/s13073-024-01368-7","article-title":"Microglial heterogeneity in the ischemic stroke mouse brain of both sexes","volume":"16","author":"Zhang","year":"2024","journal-title":"Genome Med."},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"eadl2184","DOI":"10.1126\/scitranslmed.adl2184","article-title":"Vagal stimulation ameliorates murine colitis by regulating SUMOylation","volume":"16","author":"Youssef","year":"2024","journal-title":"Sci. Transl. Med."}],"container-title":["Pharmaceuticals"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1424-8247\/18\/7\/1070\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,9]],"date-time":"2025-10-09T18:12:55Z","timestamp":1760033575000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1424-8247\/18\/7\/1070"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,7,20]]},"references-count":50,"journal-issue":{"issue":"7","published-online":{"date-parts":[[2025,7]]}},"alternative-id":["ph18071070"],"URL":"https:\/\/doi.org\/10.3390\/ph18071070","relation":{},"ISSN":["1424-8247"],"issn-type":[{"value":"1424-8247","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,7,20]]}}}