Antimicrobial Resistance Interdisciplinary Research Group, Singapore-MIT Alliance for Research and Technology, Singapore 138602, Singapore; Synthetic Biology Group, MIT Synthetic Biology Center, Massachusetts Institute of Technology (MIT), Cambridge, MA 02139, USA; Research Laboratory of Electronics, MIT, Cambridge, MA 02139, USA.
Department of Immunology and Infectious Diseases, Harvard T.H. Chan School of Public Health, Boston, MA 02115, USA.
Cell Syst. 2020 Sep 23;11(3):239-251.e7. doi: 10.1016/j.cels.2020.08.010.
Existing antibiotics are inadequate to defeat tuberculosis (TB), a leading cause of death worldwide. We sought potential targets for host-directed therapies (HDTs) by investigating the host immune response to mycobacterial infection. We used high-throughput CRISPR knockout and CRISPR interference (CRISPRi) screens to identify perturbations that improve the survival of human phagocytic cells infected with Mycobacterium bovis BCG (Bacillus Calmette-Guérin), as a proxy for Mycobacterium tuberculosis (Mtb). Many of these perturbations constrained the growth of intracellular mycobacteria. We identified over 100 genes associated with diverse biological pathways as potential HDT targets. We validated key components of the type I interferon and aryl hydrocarbon receptor signaling pathways that respond to the small-molecule inhibitors cerdulatinib and CH223191, respectively; these inhibitors enhanced human macrophage survival and limited the intracellular growth of Mtb. Thus, high-throughput functional genomic screens, by elucidating highly complex host-pathogen interactions, can serve to identify HDTs to potentially improve TB treatment.
现有的抗生素不足以对抗结核病（TB），这是全球主要的死亡原因。我们通过研究宿主对分枝杆菌感染的免疫反应，寻找宿主定向治疗（HDT）的潜在靶点。我们使用高通量 CRISPR 敲除和 CRISPR 干扰（CRISPRi）筛选来识别可改善感染牛分枝杆菌卡介苗（BCG）（卡介苗）的人吞噬细胞生存能力的扰动，作为结核分枝杆菌（Mtb）的替代物。这些扰动中的许多都限制了细胞内分枝杆菌的生长。我们鉴定了 100 多个与多种生物途径相关的基因，这些基因可能是 HDT 的潜在靶点。我们验证了对小分子抑制剂塞杜林替尼和 CH223191 分别作出反应的 I 型干扰素和芳基烃受体信号通路的关键组成部分；这些抑制剂增强了人类巨噬细胞的存活并限制了 Mtb 的细胞内生长。因此，通过阐明高度复杂的宿主-病原体相互作用的高通量功能基因组筛选，可以鉴定出潜在的 HDT，以改善结核病治疗。
