Dataset Information

Transcriptome and translatome profiling of Mtb-infected human DC identifies GSK-3beta as a molecular switch of rapamycin-driven immune stimulation

ABSTRACT: In human primary dendritic cells (DC) rapamycin - an autophagy inducer and protein synthesis inhibitor - overcomes the autophagy block induced by Mycobacterium tuberculosis (Mtb) and promotes Th1 response via IL-12 secretion. Here, the immunostimulatory activity of rapamycin in Mtb-infected DC was investigated by analyzing the transcriptome and the translatome profiles. Several differentially expressed genes (DEGs) were found in transcriptome and translatome of Mtb-infected DC, showing an additional increase in presence of rapamycin, which was instead ineffective in uninfected cells. The majority of DEGs identified in the translatome resulted also present in the transcriptome-modulated genes, suggesting that both RNAs are actively translated mRNAs. The DEG in silico analysis showed significant changes in intracellular cascades related to cytokine production, cytokine-induced signaling and immune response to pathogens. In particular, rapamycin treatment of Mtb-infected DC caused a significant enrichment of IFN-beta, IFN-lambda and IFN-stimulated genes in the polysome-associated RNA fraction. In addition, IL-12, IL-23, IL-1B, IL-6, and TNF-A were induced at RNA and protein level by rapamycin in Mtb-infected DC, while IL-10 was reduced. Interestingly, upon siRNA or pharmacologically GSK-3beta inhibition, rapamycin-driven modulation of the pro- and anti-inflammatory cytokine balance was lost, thus indicating that in Mtb-infected DC GSK-3beta functions as molecular switch for the regulation of cytokine milieu. In conclusion, our study sheds light on the molecular mechanism by which autophagy contributes to DC activation during Mtb infection and highlights that rapamycin and its analogs as well as GSK-3beta modulators are promising compounds for host-directed therapy in the control of Mtb infection.

ORGANISM(S): Homo sapiens

PROVIDER: GSE163531 | GEO | 2021/04/12

REPOSITORIES: GEO

ACCESS DATA

Similar Datasets

Project description:Analysis of Mtb infected murine macrophages derived from C57Bl/6 WT, TPL2KO, IFNARKO & TPL2IFNAR DKO mice [Set 1] Tuberculosis, caused by Mycobacterium tuberculosis (Mtb), remains a leading cause of mortality and morbidity worldwide, causing approximately 1.4 million deaths per year. Key immune components for host protection during tuberculosis include the cytokines IL?12, IL?1 and TNF??, as well as IFN?? and CD4+ Th1 cells. However, immune factors determining whether individuals control infection or progress to active tuberculosis are incompletely understood. Excess amounts of type I interferon have been linked to exacerbated disease during tuberculosis in mouse models and to active disease in patients, suggesting tight regulation of this family of cytokines is critical to host resistance. In addition, the immunosuppressive cytokine IL?10 is known to inhibit the immune response to Mtb in murine models through the negative regulation of key pro-inflammatory cytokines and the subsequent Th1 response. We show here, using a combination of transcriptomic analysis, genetics and pharmacological inhibitors that the TPL-2-ERK1/2 signaling pathway is important in mediating host resistance to tuberculosis through negative regulation of type I interferon production. The TPL-2-ERK1/2 signalling pathway regulated production by macrophages of several cytokines important in the immune response to Mtb as well as regulating induction of a large number of additional genes, many in a type I IFN dependent manner. In the absence of TPL-2 in vivo, excess type I interferon promoted IL-10 production and exacerbated disease. These findings describe an important regulatory mechanism for controlling tuberculosis and reveal mechanisms by which type I interferon may promote susceptibility to this important disease. Macrophages were derived from C57Bl/6 bone marrow, plated and infected with Mtb H37Rv (or not) in duplicate wells. Samples were then harvested for RNA at time 0 (uninfected only), 15m, 30m, 1hr, 3hr, 6hr and 24hr.

Dataset Information

Transcriptome and translatome profiling of Mtb-infected human DC identifies GSK-3beta as a molecular switch of rapamycin-driven immune stimulation

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure