Macrophages exhibit remarkable functional plasticity, a requirement for their central role in tissue homeostasis. During chronic inflammation, macrophages acquire sustained inflammatory ‘states’ that contribute to disease, but there is limited understanding of the regulatory mechanisms that drive their generation. Here we describe a systematic functional genomics approach that combines genome-wide phenotypic screening in primary murine macrophages with transcriptional and cytokine profiling of genetic perturbations in primary human macrophages to uncover regulatory circuits of inflammatory states. This process identifies regulators of five distinct states associated with key features of macrophage function. Among these regulators, loss of the N⁶-methyladenosine (m6A) writer components abolishes m6A modification of TNF transcripts, thereby enhancing mRNA stability and TNF production associated with multiple inflammatory pathologies. Thus, phenotypic characterization of primary murine and human macrophages describes the regulatory circuits underlying distinct inflammatory states, revealing post-transcriptional control of TNF mRNA stability as an immunosuppressive mechanism in innate immunity.

巨噬细胞表现出显着的功能可塑性，这是它们在组织稳态中发挥核心作用的要求。在慢性炎症期间，巨噬细胞获得导致疾病的持续炎症“状态”，但对驱动其生成的调节机制的了解有限。在这里，我们描述了一种系统的功能基因组学方法，该方法将原代小鼠巨噬细胞中的全基因组表型筛选与原代人巨噬细胞中遗传扰动的转录和细胞因子分析相结合，以揭示炎症状态的调节回路。该过程识别与巨噬细胞功能的关键特征相关的五种不同状态的调节因子。在这些调节因子中，N⁶-甲基腺苷 （m6A） 写入组分的缺失消除了 TNF 转录本的 m6A 修饰，从而增强了与多种炎症相关的 mRNA 稳定性和 TNF 产生。因此，原代小鼠和人巨噬细胞的表型特征描述了不同炎症状态背后的调节回路，揭示了 TNF mRNA 稳定性的转录后控制作为先天免疫中的免疫抑制机制。