Itaconate regulates macrophage function through stressful iron‐sulfur cluster disrupting and iron metabolism rebalancing

Lipopolysaccharide (LPS)‐stimulated macrophages express an aconitate decarboxylase (IRG1, also called ACOD1), leading to accumulation of the endogenous metabolite itaconate. However, the precise mechanisms by which elevated itaconate levels alter macrophage function are not clear. Our hypothesis is itaconate affects macrophage function through some uncertain mechanism. Based on this, we established a transcriptional and proteomic signature of macrophages stimulated by itaconate and identified the pathways of IL‐1β secretion and altered iron metabolism. Consistently, the effect of IRG1 deficiency on IL‐1β secretion and iron metabolism was confirmed in IRG1 knockout THP‐1 cell lines. Several common inhibitors and other compounds were used to examine the molecular mechanisms involved. Only cysteine and antioxidants (catechin hydrate) could inhibit caspase‐1 activation and IL‐1β secretion in itaconate‐stimulated macrophages. We further found that aconitase activity was decreased by itaconate stimulation. Our results demonstrate the counteracting effects of overexpression of mitochondrial aconitase (ACO2, a tricarboxylic acid cycle enzyme) or cytosolic aconitase (ACO1, an iron regulatory protein) on IL‐1β secretion and altered iron metabolism. Both enzyme activities were inhibited by itaconate because of iron‐sulfur (Fe‐S) cluster destruction. Our findings indicate that the immunoregulatory functions of IRG1 and itaconate in macrophages are stressful Fe‐S cluster of aconitases disrupting and iron metabolism rebalancing.