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Glucocorticoids (GCs) like dexamethasone (DEX) are standard long-term SLE treatments but cause significant side effects, highlighting the need for safer steroid-sparing options. Preclinical and clinical studies suggest that dietary supplementation with omega-3 fatty acids (O3FAs), particularly docosahexaenoic acid (DHA), suppress inflammation and autoimmunity associated with SLE disease progression. We explored the steroid-sparing potential of DHA to influence suppressive effects of DEX on pathogenic gene expression.<\/jats:p>\n                <\/jats:sec>\n                <jats:sec>\n                  <jats:title>Methods<\/jats:title>\n                  <jats:p>Macrophages from SLE-prone NZBWF1 mice were first subjected to DHA (5, 10, or 25 \u00b5M), DEX (1, 10, 100, or 1000 nM), or DHA+DEX cotreatment. Following pretreatment, cells were exposed to lipopolysaccharide (LPS; 20 ng\/mL) to model SLE hyperinflammation. Effects on gene expression were analyzed by qRT-PCR and RNA-seq.<\/jats:p>\n                <\/jats:sec>\n                <jats:sec>\n                  <jats:title>Results<\/jats:title>\n                  <jats:p>\n                    qRT-PCR indicated that subinhibitory concentrations of DHA (5-10 \u00b5M) potentiated the efficacy of low-dose DEX (1-100 nM) in suppressing LPS-induced ISG expression (e.g.,\n                    <jats:italic>Irf7<\/jats:italic>\n                    ,\n                    <jats:italic>Oasl1<\/jats:italic>\n                    ,\n                    <jats:italic>Rsad2<\/jats:italic>\n                    ), amplifying the effects of DEX monotherapy by 10- to 100-fold. SynergyFinder analysis confirmed that DHA and DEX interacted synergistically in suppressing ISG expression, with significant inhibition observed at concentrations as low as 1 nM DEX and 5 \u00b5M DHA. RNA-seq revealed that combining suboptimal DHA (10 \u03bcM) and DEX (100 nM) induced 247 differentially expressed genes (DEGs) at 4 hr and 347 DEGs at 8 hr post-LPS, dramatically surpassing the effects of each treatment alone. Functional enrichment analysis indicated that DHA+DEX cotreatment robustly suppressed immune and inflammatory pathways while promoting proliferative and metabolic processes, reflecting a shift from inflammatory (M1) to pro-resolving (M2) macrophage phenotypes. DHA and DEX countered LPS effects by i) downregulating common transcription factors (TFs) canonically associated with inflammation (e.g., NF-\u03baB, AP-1, STATs, and IRF1), ii) upregulating shared regulatory factors involved in inflammation resolution (e.g., YBX1, EGR1, and BCL6), and iii) selectively influencing other regulatory factors.\n                  <\/jats:p>\n                <\/jats:sec>\n                <jats:sec>\n                  <jats:title>Discussion<\/jats:title>\n                  <jats:p>Altogether, DHA and DEX synergistically suppress inflammation by targeting common and unique molecular pathways in SLE macrophages, favoring the pro-resolving M2 phenotype. O3FA-GC cotreatment might facilitate reducing requisite steroid dosages for SLE management.<\/jats:p>\n                <\/jats:sec>","DOI":"10.1101\/2025.06.12.658906","type":"posted-content","created":{"date-parts":[[2025,6,17]],"date-time":"2025-06-17T18:20:16Z","timestamp":1750184416000},"source":"Crossref","is-referenced-by-count":0,"title":["Omega-3 Fatty Acid Synergy with Glucocorticoid in Lupus Macrophages: Targeting Pathogenic Pathways to Reduce Steroid Dependence"],"prefix":"10.64898","author":[{"given":"Lauren K.","family":"Heine","sequence":"first","affiliation":[{"name":"Department of Pharmacology and Toxicology, Michigan State University, East Lansing, MI, United States"},{"name":"Institute for Integrative Toxicology, Michigan State University, East Lansing, MI, United 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States"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4682-0824","authenticated-orcid":false,"given":"Jack R.","family":"Harkema","sequence":"additional","affiliation":[{"name":"Department of Pharmacology and Toxicology, Michigan State University, East Lansing, MI, United States"},{"name":"Institute for Integrative Toxicology, Michigan State University, East Lansing, MI, United States"},{"name":"Department of Pathobiology and Diagnostic Investigation, Michigan State University, East Lansing, MI, United States"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3441-3113","authenticated-orcid":false,"given":"Andrew J.","family":"Olive","sequence":"additional","affiliation":[{"name":"Department of Microbiology, Genetics, and Immunology, Michigan State University, East Lansing, MI, United 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