Abstract
Objective and design
Phagocytosis and clearance of apoptotic cells are essential for inflammation resolution, efficient wound healing, and tissue homeostasis. MicroRNAs are critical modulators of macrophage polarization and function. The current study aimed to investigate the role of miR-181c-5p in macrophage phagocytosis.
Materials and methods
miR-181c-5p was identified as a potential candidate in microRNA screening of RAW264.7 macrophages fed with apoptotic cells. To investigate the role of miR-181c-5p in phagocytosis, the expression of miR-181c-5p was assessed in phagocyting bone marrow-derived macrophages. Phagocytosis efficiency was measured by fluorescence microscopy. Gain- and loss-of-function studies were performed using miR-181c-5p-specific mimic and inhibitor. The expression of the phagocytosis-associated genes and proteins of interest was evaluated by RT2 profiler PCR array and western blotting, respectively.
Results
miR-181c-5p expression was significantly upregulated in the phagocyting macrophages. Furthermore, mimic-induced overexpression of miR-181c-5p resulted in the increased phagocytic ability of macrophages. Moreover, overexpression of miR-181c-5p resulted in upregulation of WAVE-2 in phagocyting macrophages, suggesting that miR-181c-5p may regulate cytoskeletal arrangement during macrophage phagocytosis.
Conclusion
Altogether, our data provide a novel function of miR-181c-5p in macrophage biology and suggest that targeting macrophage miR-181c-5p in injured tissues might improve clearance of dead cells and lead to efficient inflammation resolution.
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Acknowledgements
This work is supported, in part, by National Institutes of Health (NIH) grants HL116729 (to P.K.), HL137411 (to P.K. and J.Z.), American Heart Association Transformational Project Award 19TPA34850100 (to P.K.) and T32 Training Grant T32EB023872 (to J.H.).
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All authors contributed substantially to the work presented in this paper. JH and PK conceptualized and design the study. SS, MP, JH, PD, and SD performed the study. SS, JH, MP, and PK analyzed and organized the data. SS, JH, MP, RK, and PK interpreted the results. SS, JH, and PK were involved in writing manuscript drafts and revisions. RK and JZ provided critical appraisal and conceptual insights during the preparation of the manuscript.
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Singh, S., Henderson, J., Patil, M. et al. MicroRNA-181c-5p modulates phagocytosis efficiency in bone marrow-derived macrophages. Inflamm. Res. 71, 321–330 (2022). https://doi.org/10.1007/s00011-022-01539-5
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DOI: https://doi.org/10.1007/s00011-022-01539-5