Exploiting induced pluripotent stem cell-derived macrophages to unravel host factors influencing Chlamydia trachomatis pathogenesis

Abstract

Chlamydia trachomatis remains a leading cause of bacterial sexually transmitted infections and preventable blindness worldwide. There are, however, limited in vitro models to study the role of host genetics in the response of macrophages to this obligate human pathogen. Here, we describe an approach using macrophages derived from human induced pluripotent stem cells (iPSdMs) to study macrophage–Chlamydia interactions in vitro. We show that iPSdMs support the full infectious life cycle of C. trachomatis in a manner that mimics the infection of human blood-derived macrophages. Transcriptomic and proteomic profiling of the macrophage response to chlamydial infection highlighted the role of the type I interferon and interleukin 10-mediated responses. Using CRISPR/Cas9 technology, we generated biallelic knockout mutations in host genes encoding IRF5 and IL-10RA in iPSCs, and confirmed their roles in limiting chlamydial infection in macrophages. This model can potentially be extended to other pathogens and tissue systems to advance our understanding of host-pathogen interactions and the role of human genetics in influencing the outcome of infections.

Human Induced Pluripotent Stem Cell–Derived Macrophages for Unraveling Human Macrophage Biology

“…IPSDM can be infected with various strains of HIV-1, 34 and support Salmonella 26 and Chlamydia infection. 35 Knockout of IRF5/IL10RA by CRIPSR/Cas9 led to increased susceptibility to Chlamydia trachomatis infection in IPSDM. This approach can potentially be extended to study other pathogens to advance our understanding of mechanisms of host-pathogen interactions in macrophages and the role of human genetics in influencing the susceptibility to specific pathogens and outcome of infections.…”

Loss of IL-10 signaling in macrophages limits bacterial killing driven by prostaglandin E2

“…method and designated this line as IL-10RB comp . Separately, the IL10RA gene was knocked out (IL-10RA −/− ) in the control kolf_2 iPSC line using CRISPR/Cas9 methods (Forbester et al, 2018;Yeung et al, 2017). The IL-10RB comp and IL-10RA −/− iPSCs were differentiated into Mφs and functionally validated in an IL-10-mediated LPS suppression assay.…”

“…Detailed methods for generating IL-10RA −/− iPSCs by CRISPR/ CAS9-based genome editing in the control kolf_2 line (Yeung et al, 2017) and complementation of IL-10RB −/− iPSCs by introducing functional copies of IL-10RB gene using a TALENbased approach have been previously reported by our group (Forbester et al, 2018).…”

Human‐induced pluripotent stem cell‐derived blood products: state of the art and future directions

“…Myeloid cell differentiation toward monocytes/MФs has been performed by multiple groups, using both EB or monolayer approaches, with or without the use of a feeder-layer [87][88][89][90][91]. By contrast to iPSC-MK/erythroid differentiation, no overexpression or forward programming strategies are reported to progress iPSC-MФ differentiation.…”