M. tuberculosis infection of human iPSC-derived macrophages reveals complex membrane dynamics during xenophagy evasion
Open Access
- 1 March 2021
- journal article
- research article
- Published by The Company of Biologists in Journal of Cell Science
- Vol. 134 (5)
- https://doi.org/10.1242/jcs.252973
Abstract
Xenophagy is an important cellular defence mechanism against cytosol-invading pathogens, such as Mycobacterium tuberculosis (Mtb). Activation of xenophagy in macrophages targets Mtb to autophagosomes; however, how Mtb is targeted to autophagosomes in human macrophages at a high spatial and temporal resolution is unknown. Here, we use human induced pluripotent stem cell-derived macrophages (iPSDMs) to study the human macrophage response to Mtb infection and the role of the ESX-1 type VII secretion system. Using RNA-seq, we identify ESX-1-dependent transcriptional responses in iPSDMs after infection with Mtb. This analysis revealed differential inflammatory responses and dysregulated pathways such as eukaryotic initiation factor 2 (eIF2) signalling and protein ubiquitylation. Moreover, live-cell imaging revealed that Mtb infection in human macrophages induces dynamic ESX-1-dependent, LC3B-positive tubulovesicular autophagosomes (LC3-TVS). Through a correlative live-cell and focused ion beam scanning electron microscopy (FIB SEM) approach, we show that upon phagosomal rupture, Mtb induces the formation of LC3-TVS, from which the bacterium is able to escape to reside in the cytosol. Thus, iPSDMs represent a valuable model for studying spatiotemporal dynamics of human macrophage-Mtb interactions, and Mtb is able to evade capture by autophagic compartments. This article has an associated First Person interview with the first author of the paper.Funding Information
- Cancer Research UK (FC001092)
- Medical Research Council (FC001092)
- Wellcome Trust (FC001092)
- European Research Council (772022)
- European Respiratory Society (713406)
- H2020 Marie Sklodowska-Curie grant agreement (Project SpaTime_AnTB n° 892859)
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