Kalsum Sadaf, Xu Ruilan, Akber Mira, Huang Shengjie, Lerm Maria, Chen Yuqing, Lourda Magda, Zhou Yang, Brighenti Susanna
Center for Infectious Medicine (CIM), Department of Medicine Huddinge, Karolinska Institutet, ANA Futura, 141 52 Huddinge, Sweden.
Division of Medical Microbiology and Molecular Medicine, Department of Clinical and Experimental Medicine, Linköping University, 581 83 Linköping, Sweden.
Biomolecules. 2025 Apr 9;15(4):550. doi: 10.3390/biom15040550.
Multitarget drug discovery, including host-directed therapy, is particularly promising for tuberculosis (TB) due to the resilience of (Mtb) as well as the complexity of the host's immune response. In this proof-of-concept study, we used high-content imaging to test a novel panel of dual glycogen synthase kinase 3 beta (GSK-3β) and histone deacetylase (HDAC) 1 and 6 inhibitor candidates for their efficacy in reducing the growth of green fluorescent protein (GFP)-expressing mycobacteria in human primary macrophages. We demonstrate that all ten test compounds, also including the GSK-3β inhibitor SB415286, exhibit an antimycobacterial effect of 20-60% at low micromolar doses and are non-toxic to host cells. Mtb growth showed a positive correlation with the respective 50% inhibitory concentration (IC50) values of GSK-3β, HDAC1, and HDAC6 in each compound, indicating that compounds with a potent IC50 value for HDAC1, in particular, corresponded to higher antimycobacterial activity. Furthermore, the results from multiparametric flow cytometry and a customized multiplex RNA array demonstrated that SB415286 and selected compounds, C02 and C06, could modulate immune polarization and inflammation in Mtb-infected macrophages involving an enhanced expression of CCL2, IL-10 and S100A9, but a decrease in inflammatory mediators including COX-2, TNF-α, and NFκB. These data suggest that GSK-3β inhibition alone can decrease the intracellular growth of mycobacteria and regulate macrophage inflammation, while dual GSK-3β/HDAC inhibitors enhance this efficacy. Accordingly, the tailored design of dual GSK-3β/HDAC inhibitors could represent an innovative approach to host-directed therapy in TB.
多靶点药物发现，包括宿主导向疗法，由于结核分枝杆菌（Mtb）的抗逆性以及宿主免疫反应的复杂性，在治疗结核病（TB）方面具有特别广阔的前景。在这项概念验证研究中，我们使用高内涵成像技术，测试了一组新型的双重糖原合酶激酶3β（GSK-3β）和组蛋白去乙酰化酶（HDAC）1及6抑制剂候选物，以评估它们在减少人原代巨噬细胞中表达绿色荧光蛋白（GFP）的分枝杆菌生长方面的功效。我们证明，所有十种测试化合物，包括GSK-3β抑制剂SB415286，在低微摩尔剂量下均表现出20%-60%的抗分枝杆菌作用，且对宿主细胞无毒。Mtb的生长与每种化合物中GSK-3β、HDAC1和HDAC6各自的50%抑制浓度（IC50）值呈正相关，这表明特别是对HDAC1具有有效IC50值的化合物，其抗分枝杆菌活性更高。此外，多参数流式细胞术和定制的多重RNA阵列结果表明，SB415286以及选定的化合物C02和C06，可以调节Mtb感染的巨噬细胞中的免疫极化和炎症，包括CCL2、IL-10和S100A9表达增强，但炎症介质如COX-2、TNF-α和NFκB减少。这些数据表明，单独抑制GSK-3β可以减少分枝杆菌的细胞内生长并调节巨噬细胞炎症，而双重GSK-3β/HDAC抑制剂可增强这种功效。因此，定制设计双重GSK-3β/HDAC抑制剂可能代表了一种创新的结核病宿主导向治疗方法。
