Gentili Matteo, Carlson Rebecca J, Liu Bingxu, Hellier Quentin, Andrews Jocelyn, Qin Yue, Blainey Paul C, Hacohen Nir
Broad Institute of MIT and Harvard, Cambridge, MA, USA.
Broad Institute of MIT and Harvard, Cambridge, MA, USA; Massachusetts Institute of Technology, Department of Health Sciences and Technology, Cambridge, MA, USA.
Cell Syst. 2024 Dec 18;15(12):1264-1277.e8. doi: 10.1016/j.cels.2024.11.004. Epub 2024 Dec 9.
Stimulator of interferon genes (STING) traffics across intracellular compartments to trigger innate responses. Mutations in factors regulating this process lead to inflammatory disorders. To systematically identify factors involved in STING trafficking, we performed a genome-wide optical pooled screen (OPS). Based on the subcellular localization of STING in 45 million cells, we defined 464 clusters of gene perturbations based on their cellular phenotypes. A secondary, higher-dimensional OPS identified 73 finer clusters. We show that the loss of the gene of unknown function C19orf25, which clustered with USE1, a protein involved in Golgi-to-endoplasmic reticulum (ER) transport, enhances STING signaling. Additionally, HOPS deficiency delayed STING degradation and consequently increased signaling. Similarly, GARP/RIC1-RGP1 loss increased STING signaling by delaying STING Golgi exit. Our findings demonstrate that genome-wide genotype-phenotype maps based on high-content cell imaging outperform other screening approaches and provide a community resource for mining factors that impact STING trafficking and other cellular processes.
干扰素基因刺激因子（STING）穿梭于细胞内区室以触发先天性免疫反应。调节这一过程的因子发生突变会导致炎症性疾病。为了系统地鉴定参与STING穿梭的因子，我们进行了全基因组光学池筛选（OPS）。基于4500万个细胞中STING的亚细胞定位，我们根据细胞表型定义了464个基因扰动簇。二次的、更高维度的OPS鉴定出73个更精细的簇。我们发现，与参与高尔基体到内质网（ER）运输的蛋白质USE1聚集在一起的功能未知基因C19orf25的缺失增强了STING信号传导。此外，同型融合与蛋白分选复合物（HOPS）缺陷延迟了STING的降解，从而增加了信号传导。同样，GARP/RIC1-RGP1缺失通过延迟STING从高尔基体输出而增加了STING信号传导。我们的研究结果表明，基于高内涵细胞成像的全基因组基因型-表型图谱优于其他筛选方法，并为挖掘影响STING穿梭及其他细胞过程的因子提供了一个公共资源。
