Myeloid-specific TAK1 deletion results in reduced brain monocyte infiltration and improved outcomes after stroke

Abstract

Background: Activation of transforming growth factor-β-activated kinase 1 (TAK1) occurs after stroke and leads to an exacerbation of brain injury. TAK1 is involved in innate and adaptive immune responses, but it has divergent inflammatory effects that are dependent on the cell type in which it is activated. There is a robust infiltration of myeloid cells after stroke; however, the contribution of myeloid TAK1 to cerebral ischemia is currently unknown. We hypothesized that myeloid-specific deletion of TAK1 would protect against ischemic brain injury.

Methods: Myeloid TAK1^ΔM and wild-type (WT) mice were subjected to middle cerebral artery occlusion (MCAo). Brain-infiltrating and splenic immune cells were evaluated at 3 days after stroke. Assessment of infarct size and behavioral deficits were performed on days 3 and 7 post-stroke.

Results: Infarcts were significantly smaller in TAK1^ΔM mice (p < 0.01), and behavioral deficits were less severe despite equivalent reduction in cerebral blood flow. Flow cytometry demonstrated an increase in the frequency of splenic monocytes and neutrophils (p < 0.05) and a decrease in splenic CD3⁺ T (p < 0.01) and CD19⁺ B (p = 0.06) cells in TAK1^ΔM mice compared to WT at baseline. Three days after stroke, a significant increase in the number of brain-infiltrating immune cell was observed in both TAK1^ΔM (p < 0.05) and WT (p < 0.001) mice compared to their respective shams. However, there was a significant decrease in the infiltrating CD45^hi immune cell counts (p < 0.05), with a pronounced reduction in infiltrating monocytes (p < 0.001) in TAK1^ΔM after stroke compared to WT stroke mice. Additionally, a significant reduction in CD49d⁺ monocytes was seen in the brains of TAK1^ΔM stroke mice compared to wild-type mice. Importantly, TAK1^ΔM MCAo mice had smaller infarcts and improved behavioral outcomes at day 7 post-stroke.

Conclusion: Our results showed that deletion of myeloid TAK1 resulted in smaller infarcts and improved functional outcomes at the peak of inflammation (day 3) and a reduction in brain-infiltrating immune cells that were primarily monocytes. Myeloid TAK1 deletion was also protective at 7 days post MCAo, reflecting a detrimental role of myeloid TAK1 in the progression of ischemic injury.

Keywords: Inflammation; Ischemic stroke; Monocytes; Neutrophils; TAK1.