Ferroptotic cardiomyocyte-derived exosomes promote cardiac macrophage M1 polarization during myocardial infarction

Abstract

Ferroptosis is a mode of cell death that occurs in myocardial infarction (MI). Signals emanating from apoptotic cells are able to induce macrophage polarization through exosome-loading cargos, which plays a vital role in the process of disease. However, whether ferroptotic cardiomyocytes derived exosome (MI-Exo) during MI act on macrophage polarization and its mechanism remain unclear. In this study, a MI mouse model was established, and cardiac function evaluation and pathological staining were performed. The effect of MI-Exo on polarization of RAW264.7 cells was assessed by the expression of IL-10 and NOS2. Ferroptosis inhibitor of ferrostatin-1 was used to verify whether MI-Exo function was dependents on ferroptosis. Cardiac function and myocardial histomorphology were markedly impaired and massive immune cell infiltration in MI mice, compared with the sham group. The significantly increased MDA content and Fe2+ accumulation in the heart tissue of MI mice suggested cardiomyocyte ferroptosis. Compared with the sham group, the expression of M1 marker NOS2 was significantly up-regulated and M2 marker IL-10 was significantly down-regulated in the heart tissue of MI mice. Exosome-derived from MI HL-1 cell-treated with ferrostatin-1 (Fer-1-Exo) and MI-Exo were internalized by RAW 264.7 cells. Compared with culture alone, co-cultured with MI-Exo significantly promoted NOS2 expression and suppressed IL-10 expression, and decreased proportion of Arginase-1-labeled M2 macrophages, also inhibited phagocytosis of RAW 264.7 cells. Wnt1 and β-cantenin expression also elevated after treated with MI-Exo. However, co-cultured with Fer-1-Exo significantly reversed the above changes on RAW 264.7 cells induced by MI-Exo. In conclusion, ferroptotic cardiomyocytes-derived exosome crosstalk macrophage to induce M1 polarization via Wnt/β-cantenin pathway, resulting in pathological progress in MI. This understanding provides novel therapeutic target for MI.

Ischemia-reperfusion injury: molecular mechanisms and therapeutic targets

“…This activation of the canonical pathway plays a beneficial role in injured organs through various processes such as inflammation, ECM remodeling, angiogenesis, fibrosis, and nerve regeneration. Within different organs, 49 , 120 – 122 , 134 , 135 , 146 , 499 – 501 it serves as a compensatory protective response aimed at mitigating damage caused by ischemia and promoting the repair of resulting injuries. However, when these mechanisms become decompensated, corresponding pathological processes occur.…”

“…The authors suggested that cardiomyocyte ferroptosis occurs during the ischemia phase, accompanied by macrophage polarization toward the M1 phenotype and verified that hypoxic HL-1 cells undergo ferroptosis in vitro and the exosomes derived from ferroptotic HL-1 cells induce M1 macrophage transformation in RAW264.7 cells by activating the Wnt/β-catenin pathway. 120

Fig. 3 Wnt signaling pathway and targeted therapy in ECM remodeling, angiogenesis, cardiac hypertrophy, and fibrosis during myocardial I/R injury.

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Development of a prognostic model based on ferroptosis-related genes for colorectal cancer patients and exploration of the biological functions of NOS2 in vivo and in vitro

“…NOS2 is an inducible isoform of NOS enzymes and functions as a key inflammatory enzyme responsible for nitric oxide biosynthesis ( 33 ). Recent studies connected NOS2 and ferroptosis were almost based on bioinformatic analysis, and NOS2 had been identified as the marker of ferroptosis functions in the process of HIF-1 signaling pathway, NOD-like receptor signaling pathway, central carbon metabolism and macrophage polarization ( 13 , 34 , 35 ). The dysregulation of NOS2 expression can be observed under pathological conditions, including cytokine exposure, inflammation and tumors ( 33 , 36 ).…”

Effect of hypercholesterolemia on circulating and cardiomyocyte-derived extracellular vesicles

“…Earlier studies showed that CM EVs activate monocytes in various disease conditions, such as in angiotensin II-induced hypertrophy, thereby contributing to inflammation 21 , 22 . In contrast, in hypoxic conditions, EVs promote macrophage polarization to the reparative M2 phenotype 23 , 24 . These results suggest that CM EVs might play a role in HC-induced inflammation.…”