Dual GSK-3β/HDAC Inhibitors Enhance the Efficacy of Macrophages to Control <i>Mycobacterium tuberculosis</i> Infection

Multitarget drug discovery, including host-directed therapy, is particularly promising for tuberculosis (TB) due to the resilience of <i>Mycobacterium tuberculosis</i> (Mtb) as well as the complexity of the host’s immune response. In this proof-of-concept study, we used high-content imag...

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Main Authors: Sadaf Kalsum, Ruilan Xu, Mira Akber, Shengjie Huang, Maria Lerm, Yuqing Chen, Magda Lourda, Yang Zhou, Susanna Brighenti
Format: Article
Language:English
Published: MDPI AG 2025-04-01
Series:Biomolecules
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Online Access:https://www.mdpi.com/2218-273X/15/4/550
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author Sadaf Kalsum
Ruilan Xu
Mira Akber
Shengjie Huang
Maria Lerm
Yuqing Chen
Magda Lourda
Yang Zhou
Susanna Brighenti
author_facet Sadaf Kalsum
Ruilan Xu
Mira Akber
Shengjie Huang
Maria Lerm
Yuqing Chen
Magda Lourda
Yang Zhou
Susanna Brighenti
author_sort Sadaf Kalsum
collection DOAJ
description Multitarget drug discovery, including host-directed therapy, is particularly promising for tuberculosis (TB) due to the resilience of <i>Mycobacterium tuberculosis</i> (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.
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spelling doaj-art-1d2a2a4053e64e26be35a5b1f4477c342025-08-20T03:14:17ZengMDPI AGBiomolecules2218-273X2025-04-0115455010.3390/biom15040550Dual GSK-3β/HDAC Inhibitors Enhance the Efficacy of Macrophages to Control <i>Mycobacterium tuberculosis</i> InfectionSadaf Kalsum0Ruilan Xu1Mira Akber2Shengjie Huang3Maria Lerm4Yuqing Chen5Magda Lourda6Yang Zhou7Susanna Brighenti8Center for Infectious Medicine (CIM), Department of Medicine Huddinge, Karolinska Institutet, ANA Futura, 141 52 Huddinge, SwedenCenter for Infectious Medicine (CIM), Department of Medicine Huddinge, Karolinska Institutet, ANA Futura, 141 52 Huddinge, SwedenCenter for Infectious Medicine (CIM), Department of Medicine Huddinge, Karolinska Institutet, ANA Futura, 141 52 Huddinge, SwedenState Key Laboratory of Bioactive Molecules and Druggability Assessment, Guangdong Basic Research Center of Excellence for Natural Bioactive Molecules and Discovery of Innovative Drugs, International Cooperative Laboratory of Traditional Chinese Medicine Modernization and Innovative Drug Discovery of Chinese Ministry of Education School of Pharmacy, Jinan University, 855 Xingye Avenue, Guangzhou 510632, ChinaDivision of Medical Microbiology and Molecular Medicine, Department of Clinical and Experimental Medicine, Linköping University, 581 83 Linköping, SwedenState Key Laboratory of Bioactive Molecules and Druggability Assessment, Guangdong Basic Research Center of Excellence for Natural Bioactive Molecules and Discovery of Innovative Drugs, International Cooperative Laboratory of Traditional Chinese Medicine Modernization and Innovative Drug Discovery of Chinese Ministry of Education School of Pharmacy, Jinan University, 855 Xingye Avenue, Guangzhou 510632, ChinaDivision of Clinical Microbiology, Department of Laboratory Medicine, Karolinska Institutet, ANA Futura, 141 52 Huddinge, SwedenState Key Laboratory of Bioactive Molecules and Druggability Assessment, Guangdong Basic Research Center of Excellence for Natural Bioactive Molecules and Discovery of Innovative Drugs, International Cooperative Laboratory of Traditional Chinese Medicine Modernization and Innovative Drug Discovery of Chinese Ministry of Education School of Pharmacy, Jinan University, 855 Xingye Avenue, Guangzhou 510632, ChinaCenter for Infectious Medicine (CIM), Department of Medicine Huddinge, Karolinska Institutet, ANA Futura, 141 52 Huddinge, SwedenMultitarget drug discovery, including host-directed therapy, is particularly promising for tuberculosis (TB) due to the resilience of <i>Mycobacterium tuberculosis</i> (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.https://www.mdpi.com/2218-273X/15/4/550tuberculosis<i>Mycobacterium tuberculosis</i>macrophagehost-directed therapycompoundglycogen synthase kinase 3 beta (GSK-3β)
spellingShingle Sadaf Kalsum
Ruilan Xu
Mira Akber
Shengjie Huang
Maria Lerm
Yuqing Chen
Magda Lourda
Yang Zhou
Susanna Brighenti
Dual GSK-3β/HDAC Inhibitors Enhance the Efficacy of Macrophages to Control <i>Mycobacterium tuberculosis</i> Infection
Biomolecules
tuberculosis
<i>Mycobacterium tuberculosis</i>
macrophage
host-directed therapy
compound
glycogen synthase kinase 3 beta (GSK-3β)
title Dual GSK-3β/HDAC Inhibitors Enhance the Efficacy of Macrophages to Control <i>Mycobacterium tuberculosis</i> Infection
title_full Dual GSK-3β/HDAC Inhibitors Enhance the Efficacy of Macrophages to Control <i>Mycobacterium tuberculosis</i> Infection
title_fullStr Dual GSK-3β/HDAC Inhibitors Enhance the Efficacy of Macrophages to Control <i>Mycobacterium tuberculosis</i> Infection
title_full_unstemmed Dual GSK-3β/HDAC Inhibitors Enhance the Efficacy of Macrophages to Control <i>Mycobacterium tuberculosis</i> Infection
title_short Dual GSK-3β/HDAC Inhibitors Enhance the Efficacy of Macrophages to Control <i>Mycobacterium tuberculosis</i> Infection
title_sort dual gsk 3β hdac inhibitors enhance the efficacy of macrophages to control i mycobacterium tuberculosis i infection
topic tuberculosis
<i>Mycobacterium tuberculosis</i>
macrophage
host-directed therapy
compound
glycogen synthase kinase 3 beta (GSK-3β)
url https://www.mdpi.com/2218-273X/15/4/550
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