Human pluripotent stem cell-derived macrophages host Mycobacterium abscessus infection
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ABSTRACT:
PROVIDER: PRJNA762242 | ENA |
REPOSITORIES: ENA
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Project description:Human macrophages are a natural host of many mycobacterium species, including Mycobacterium abscessus (M. abscessus), an emerging pathogen affecting patients with lung diseases and immunocompromised individuals. There are few available treatments and the search for effective antibiotics against M. abscessus has been hindered by the lack of a tractable in vitro intracellular model of infection. Here, we established a reliable model for M. abscessus infection using human pluripotent stem cell-derived macrophages (hPSC-macrophages). hPSC differentiation permitted a reproducible generation of functional human macrophages that were highly susceptible to M. abscessus infection. Electron microscopy demonstrated that M. abscessus was present in the vacuoles of hPSC-macrophages. RNA-sequencing analysis revealed a time dependent host cell response to M. abscessus, with differing gene and protein expression patterns observed at 3-hours, 24-hours and 48-hours post-infection. Culture of engineered tdTOMATO-expressing hPSC-macrophages with GFP-expressing M. abscessus enabled rapid and image-based high-throughput analysis of intracellular infection and quantitative assessment of antibiotic resistance and efficacy. Our study describes the first hPSC-based model for M. abscessus infection, which represents a novel platform for studying M. abscessus-host interaction and an accessible tool for drug discovery.
2022-08-10 | GSE207456 | GEO
Project description:Generation and analysis of human macrophages from induced pluripotent stem cells were infected by Mycobacterium abscessus to examine susceptibility and transcriptomic response
2022-08-10 | GSE183865 | GEO
Project description:Human pluripotent stem cell-derived macrophages host Mycobacterium abscessus infection
| PRJNA855683 | ENA
Project description:The initial interactions between a host cell and a pathogen can dictate disease outcomes and are important targets for host-directed therapies. Mycobacterium abscessus (Mab) is a highly antibiotic resistant, rapidly growing non-tuberculous mycobacterium that infects patients with chronic lung diseases, like cystic fibrosis. Mab can infect host immune cells, such as macrophages, which contribute to its pathogenesis. However, our understanding of host genes that modulate the initial host-Mab interactions remains unclear. Here, we developed a functional genetic approach by coupling a Mab fluorescent reporter with a genome-wide knockout library in murine macrophages. We used this approach to conduct a forward genetic screen to uncover host genes that contribute to the uptake of Mab by macrophages. We identified known regulators of phagocytosis, such as the integrin Itgb2, and uncovered a key requirement for glycosaminoglycan (sGAG) synthesis for macrophages to efficiently bind to Mab. CRISPR-Cas9 targeting of three key sGAG biosynthesis regulators, UGDH, B4GALT7 and B3GAT3 resulted in reduced uptake of both smooth and rough Mab variants by macrophages. Mechanistic studies suggest that sGAGs function upstream of pathogen engulfment and are required for uptake of Mab but not Escherichia coli or latex beads. Further investigation found that the loss of sGAGs reduces the surface expression, but not the mRNA expression, of key integrins, suggesting a key role for sGAGs in modulating surface receptor availability. Together these studies globally defined and characterized important regulators of macrophage-Mab interactions and are a first step better understand important host genes that contribute to Mab pathogenesis and disease.
2023-01-13 | GSE221413 | GEO
Project description:RNA sequencing (RNA-seq) of Mycobacterium abscessus in four infection-relevant culture conditions: hypoxic stress, artificial sputum medium, kanamycin-treated medium, and erythromycin-treated medium. Triplicate cultures of M. abscessus were grown in (1) Artificial Sputum media, (2) hypoxic conditions, (3) the presence of kanamycin, and (4) the presence of erythromycin. Triplicate controls were prepared for sample (1) and samples (2-4).
2016-03-02 | E-GEOD-78787 | biostudies-arrayexpress
Project description:Mycobacterium abscessus (Mab) infection can be a deadly infection in patients with chronic lung disease like cystic fibrosis. In vitro and in vivo, Mab may adopt a smooth (S) or rough (R) morphotype, the latter with fewer cell wall glycopeptidolipids. Although there are studies of the host response against constitutive Mab-S and Mab-R, we report the first analysis of early transcriptional events and responses in mouse bone marrow derived macrophages (BMDMs) upon infection with our media-selected, isogenic, interchangeable Mab-S and Mab-R. The early transcriptional events after infection with both the morphotypes showed considerable overlap of the pro-inflammatory genes that were differentially regulated compared to the uninfected macrophages. However, we also observed some signature genes which were upregulated during Mab-S compared to Mab-R infections. Media selected Mab-S and Mab-R behave in a similar fashion to constitutive S and R types with respect to pathogenesis and immune response and will likely be a better model of in vivo infection than multiply passaged, mutated organisms.
2023-01-31 | GSE221966 | GEO
Project description:We report the transcriptome of M. abscessus in amoebae and macrophages. M. abscessus intra-amoebal and intra-macrophagic transcriptomes demonstrate of the potential of M. abscessus to adapt to an intracellular lifestyle, though amoebae largely contribute to the enhancement of M. abscessus survival macrophages.
2019-09-20 | GSE124972 | GEO
Project description:RNA sequencing (RNA-seq) of Mycobacterium abscessus in four infection-relevant culture conditions: hypoxic stress, artificial sputum medium, kanamycin-treated medium, and erythromycin-treated medium.
2016-03-02 | GSE78787 | GEO
Project description:Mycobacterium abscessus is nowadays under the spotlight of the scientific community. This pathogenic mycobacteria is indeed responsible for a wide spectrum of infections involving mostly pulmonary infections in patients with cystic fibrosis. M. abscessus is intrinsically resistant to a broad range of antibiotics, including most antitubercular drugs, and is considered the most pathogenic and chemotherapy-resistant rapidly growing mycobacterium. Consequently, with very limited treatment options, the development of new therapeutic approaches to fight this pathogen are urgently needed. In this context, 19 oxadiazolone (OX) derivatives have been investigated for their antibacterial activity against both the rough (R) and smooth (S) variants of M. abscessus. Several OXs were active against extracellular M. abscessus growth with moderated minimal inhibitory concentrations (MIC), or intracellularly by inhibiting M. abscessus growth inside infected macrophages with MIC values similar to those of imipenem. Such promising results prompted us to identify the potential target enzymes of the sole extra and intracellular inhibitor of M. abscessus growth, i.e., iBpPPOX via activity-based protein profiling combined with mass spectrometry. This approach led to the identification of 21 potential protein candidates being mostly involved in M. abscessus lipid metabolism and/or in cell wall biosynthesis.
2020-08-14 | PXD015680 | Pride
Project description:Mycobacterium abscessus is nowadays under the spotlight of the scientific community. This pathogenic mycobacteria is indeed responsible for a wide spectrum of infections involving mostly pulmonary infections in patients with cystic fibrosis. M. abscessus is intrinsically resistant to a broad range of antibiotics, including most antitubercular drugs, and is considered the most pathogenic and chemotherapy-resistant rapidly growing mycobacterium. Consequently, with very limited treatment options, the development of new therapeutic approaches to fight this pathogen are urgently needed. 38 new analogs of Cyclipostins & Cyclophostin (CyC), compounds naturally produced by Streptomyces species, have been synthesized. Their antibacterial activities against clinical isolates belonging to the M. chelonae-abscessus clade, as well as Gram-negative and Gram-positive bacteria have been evaluated by the REMA method. The intracellular activities of the CyC against intramacrophagic M. abscessus have also been investigated and compared to those of imipenem. The CyCs displayed very low toxicity towards host cells and their inhibitory activity was exclusively restricted to mycobacteria. The best candidate, CyC17, showed a high selectivity for mycobacteria with MIC values (<2 up to 40 µg/mL) comparable to those of most classical antibiotics used to treat M. abscessus infections. Of importance, several CyCs were active against extracellular M. abscessus growth (i.e., CyC17 / CyC18β / CyC25 / CyC26) or against intracellular mycobacteria inside macrophages (i.e., CyC7α,β / CyC8α,β) with MIC values similar to or better than those of standard antibiotics. Based on these results, we intended to identify the potential target enzymes of CyC17/CyC26 in M. abscessus by activity-based protein profiling (ABPP) approach coupled with mass spectrometry differential analysis.
2019-10-08 | PXD014255 | Pride