Abstract Body: Macrophages (Mφ) play central roles in inducing the robust inflammatory response to acute ischemia and promoting anti-inflammatory actions necessary for myocardial repair. The broad range of functional repertoire stems from Mφ plasticity dictated by their interaction with the dynamic ischemic milieu. The limited understanding of such interactions contributes to the lack of means to mitigate inflammatory injury in MI patients. We hypothesize that the dynamic Mφ plasma membrane-mediated process macropinocytosis (MP) is essential for Mφ’s activation and function evolvement in ischemia. By MP, Mφs engulf large amounts of soluble contents from the ischemic environment, and the macropinosomes formed from MP become signaling hubs of DAMP (Damage-Associated Molecular Pattern)-PRR (Pattern Recognition Receptor). We aim to determine if ischemia activates MP in Mφs, the signaling pathways that MP promotes, and the role of this process in post-ischemia repair. The rationale is based on our data demonstrating only soluble DAMP (sDAMP), not tissue debris, activates Mφs. We tested our hypothesis using the murine MI model and bone marrow-derived Mφs (BMDM) and used unpaired t-tests for statistical analyses. We uncover that ischemia markedly increases MP in Mφs; blocking MP by the inhibitors or siRNA-mediated knock-down of the Na+H+ exchanger abolished the production of potent cytokines such as IL1β from Mφs in vitro and in vivo. Furthermore, MP activates NF-κB and MAP kinase triggered by sDAMP and ischemia, critical pathways to produce inflammatory cytokine and chemokine. Intriguingly, mTORC1 activation in BMDM and Mφs in the ischemic myocardium depends on MP. These data suggest MP organizes the activation signal and the pathway required for metabolic rewiring in inflammatory Mφs. Suppressing MP at the earlier stage of ischemia (72h) when sDAMP is abundant in the ischemic tissue significantly reduces mortality and improves cardiac function. In conclusion, we identify MP functionalizes the danger signals in bulk and integrates activation signals with metabolic rewiring to promote and sustain the inflammatory function. Targeting MP is better equipped to reduce inflammatory injury as it modifies the dynamic interaction of Mφs with the ischemic environment.