Kuijl Coenraad, Savage Nigel D L, Marsman Marije, Tuin Adriaan W, Janssen Lennert, Egan David A, Ketema Mirjam, van den Nieuwendijk Rian, van den Eeden Susan J F, Geluk Annemieke, Poot Alex, van der Marel Gijs, Beijersbergen Roderick L, Overkleeft Hermen, Ottenhoff Tom H M, Neefjes Jacques
Division of Tumor Biology, The Netherlands Cancer Institute, Plesmanlaan 121, 1066 CX Amsterdam, The Netherlands.
Nature. 2007 Nov 29;450(7170):725-30. doi: 10.1038/nature06345.
With the emergence of multidrug resistant (MDR) bacteria, it is imperative to develop new intervention strategies. Current antibiotics typically target pathogen rather than host-specific biochemical pathways. Here we have developed kinase inhibitors that prevent intracellular growth of unrelated pathogens such as Salmonella typhimurium and Mycobacterium tuberculosis. An RNA interference screen of the human kinome using automated microscopy revealed several host kinases capable of inhibiting intracellular growth of S. typhimurium. The kinases identified clustered in one network around AKT1 (also known as PKB). Inhibitors of AKT1 prevent intracellular growth of various bacteria including MDR-M. tuberculosis. AKT1 is activated by the S. typhimurium effector SopB, which promotes intracellular survival by controlling actin dynamics through PAK4, and phagosome-lysosome fusion through the AS160 (also known as TBC1D4)-RAB14 pathway. AKT1 inhibitors counteract the bacterial manipulation of host signalling processes, thus controlling intracellular growth of bacteria. By using a reciprocal chemical genetics approach, we identified kinase inhibitors with antibiotic properties and their host targets, and we determined host signalling networks that are activated by intracellular bacteria for survival.
随着多重耐药(MDR)细菌的出现,开发新的干预策略势在必行。目前的抗生素通常靶向病原体而非宿主特异性生化途径。在此,我们开发了激酶抑制剂,可阻止鼠伤寒沙门氏菌和结核分枝杆菌等不相关病原体在细胞内生长。利用自动显微镜对人类激酶组进行的RNA干扰筛选揭示了几种能够抑制鼠伤寒沙门氏菌细胞内生长的宿主激酶。所鉴定的激酶聚集在围绕AKT1(也称为PKB)的一个网络中。AKT1抑制剂可阻止包括多重耐药结核分枝杆菌在内的各种细菌在细胞内生长。AKT1由鼠伤寒沙门氏菌效应蛋白SopB激活,SopB通过PAK4控制肌动蛋白动力学,并通过AS160(也称为TBC1D4)-RAB14途径促进吞噬体-溶酶体融合,从而促进细胞内存活。AKT1抑制剂可抵消细菌对宿主信号传导过程的操纵,从而控制细菌在细胞内的生长。通过使用相互化学遗传学方法,我们鉴定了具有抗生素特性的激酶抑制剂及其宿主靶点,并确定了被细胞内细菌激活以实现存活的宿主信号网络。