Generation of Myeloid-Specific Bmal1 Knockout Mice and Identification of Bmal1-Regulated Ferroptosis in Macrophages

Circadian clocks have a fundamental role in many physiological processes. Bmal1 (basic helix-loop-helix ARNT like 1) is a central master circadian clock gene. The global Bmal1 knockout mice were shown to have a loss of circadian rhythms, acceleration of aging, and shortened life span. However, global Bmal1 knockout mice did not exactly reflect the Bmal1 function in specific cell or tissue types. To address the importance of circadian rhythms in macrophages, we generated myeloid-specific Bmal1 knockout mice. The efficacy of Bmal1 gene deletion in macrophages was identified at DNA, transcription, protein levels, and function. In contrast to global Bmal1 knockout mice, Bmal1^flox/flox and Bmal1^mye-/- mice did not exhibit aging phenotypes. However, the deletion of Bmal1 resulted in a loss of rhythmic expression of the circadian genes in macrophages. RNA-Seq revealed that Bmal1 regulated the expression of cell death-related genes in macrophages. Furthermore, these genes have been identified as clock-controlled genes in rhythmic cell models, and Bmal1 controlled the rhythmic expression of these genes in macrophages. Finally, Bmal1 inhibited RSL3-induced ferroptosis in macrophages through Phgdh. In summary, the model of myeloid-specific Bmal1 knockout mice was successfully constructed, providing a tool for the study of the roles of Bmal1 in macrophages and the peripheral circadian clock. Meanwhile, Bmal1 regulates ferroptosis in macrophages.