The ability of Mycobacterium tuberculosis (Mtb) to establish latency affects disease and response to treatment. The host factors that influence the establishment of latency remain elusive. We engineered a multi-fluorescent Mtb strain that reports survival, active replication, and stressed non-replication states and determined the host transcriptome of the infected macrophages in these states. Additionally, we conducted a genome-wide CRISPR screen to identify host factors that modulated the phenotypic state of Mtb. We validated hits in a phenotype-specific manner and prioritized membrane magnesium transporter 1 (MMGT1) for a detailed mechanistic investigation. Mtb infection of MMGT1-deficient macrophages promoted a switch to persistence, upregulated lipid metabolism genes, and accumulated lipid droplets during infection. Targeting triacylglycerol synthesis reduced both droplet formation and Mtb persistence. The orphan G protein-coupled receptor GPR156 is a key inducer of droplet accumulation in ΔMMGT1 cells. Our work uncovers the role of MMGT1-GPR156-lipid droplets in the induction of Mtb persistence.

结核分枝杆菌( Mtb ) 建立潜伏期的能力会影响疾病和治疗反应。影响潜伏期建立的宿主因素仍然难以捉摸。我们设计了一种多荧光Mtb菌株，可以报告存活、活跃复制和应激非复制状态，并确定这些状态下受感染巨噬细胞的宿主转录组。此外，我们进行了全基因组CRISPR 筛选，以确定调节Mtb表型状态的宿主因子。我们以表型特异性方式验证了命中，并优先考虑膜镁转运蛋白 1 (MMGT1) 进行详细的机制研究。 MMGT1缺陷型巨噬细胞的Mtb感染促进了向持久性的转变，上调脂质代谢基因，并在感染过程中积累脂滴。以三酰甘油合成为目标可减少液滴形成和结核分枝杆菌持久性。孤儿 G 蛋白偶联受体 GPR156 是ΔMMGT1细胞中液滴积累的关键诱导物。我们的工作揭示了 MMGT1-GPR156-脂滴在诱导Mtb持久性中的作用。