Human iPSC-derived alveolar macrophages reveal macrophage subtype functions of itaconate in M. tuberculosis defense
Adam S. Krebs, Tomi Lazarov, Anthony T. Reynolds, Kimberly A. Dill-McFarland, Abigail Xie, James M. Bean, Muxue Du, Olivier Levy, John A. Buglino, Aaron Zhong, Anna-Lena Neehus, Stéphanie Boisson-Dupuis, Jean-Laurent Casanova, Elouise E. Kroon, Marlo Möller, Thomas R. Hawn, Ting Zhou, Lydia W.S. Finley, Marc Antoine Jean Juste, Dan W. Fitzgerald, Frederic Geissmann, Michael S. Glickman
Adam S. Krebs, Tomi Lazarov, Anthony T. Reynolds, Kimberly A. Dill-McFarland, Abigail Xie, James M. Bean, Muxue Du, Olivier Levy, John A. Buglino, Aaron Zhong, Anna-Lena Neehus, Stéphanie Boisson-Dupuis, Jean-Laurent Casanova, Elouise E. Kroon, Marlo Möller, Thomas R. Hawn, Ting Zhou, Lydia W.S. Finley, Marc Antoine Jean Juste, Dan W. Fitzgerald, Frederic Geissmann, Michael S. Glickman
View: Text | PDF
Research Article Immunology Infectious disease Pulmonology

Human iPSC-derived alveolar macrophages reveal macrophage subtype functions of itaconate in M. tuberculosis defense

Abstract

Mycobacterium tuberculosis (Mtb) survives within multiple macrophage populations during infection, including alveolar macrophages (AMs) and recruited inflammatory macrophages. In mice, itaconate, produced in macrophages by ACOD1-mediated decarboxylation of aconitate, has direct antimicrobial activity, modulates inflammatory cytokines, and is required for resistance to Mtb infection. The role of itaconate in human macrophages is less clear, and it is unknown whether itaconate mediates distinct effects in macrophage subtypes. Here, we investigated the role of itaconate in macrophages derived from human induced pluripotent stem cells (iPSCs), induced by either GM-CSF to resemble AMs (AM-like cells, hereafter ipAM-Ls) or M-CSF to resemble monocyte-derived macrophages (MDM-like cells, hereafter ipMDM-Ls). Both human macrophage types produced substantially less itaconate than mouse macrophages, and ipAM-Ls produced 4-fold less itaconate than ipMDM-Ls. Surprisingly, ACOD1-deficient ipAM-Ls, but not ipMDM-Ls, were permissive for Mtb growth. Moreover, itaconate functioned to dampen the Mtb-induced inflammatory response in ipMDM-Ls, but not ipAM-Ls, affecting both the type I IFN and TNF pathways. These results indicate that itaconate is involved in human macrophage responses to tuberculosis, with distinct roles in different macrophage subsets. These results also show that genetically tractable iPSC-derived macrophages are a useful model to dissect cellular host-pathogen interactions in human macrophages.

Authors

Adam S. Krebs, Tomi Lazarov, Anthony T. Reynolds, Kimberly A. Dill-McFarland, Abigail Xie, James M. Bean, Muxue Du, Olivier Levy, John A. Buglino, Aaron Zhong, Anna-Lena Neehus, Stéphanie Boisson-Dupuis, Jean-Laurent Casanova, Elouise E. Kroon, Marlo Möller, Thomas R. Hawn, Ting Zhou, Lydia W.S. Finley, Marc Antoine Jean Juste, Dan W. Fitzgerald, Frederic Geissmann, Michael S. Glickman

×

Figure 6

Human macrophages are feeble producers of itaconate with Mtb infection.

Options: View larger image (or click on image) Download as PowerPoint
Human macrophages are feeble producers of itaconate with Mtb infection. ...
(A and B) Expression of ACOD1 protein in iPSC-derived macrophage subtypes. Anti-ACOD1/Vinculin immunoblot of ipMDM-L or ipAM-L cells after 6 hours of stimulation with LPS (100 ng/mL) (A) and normalized quantitation of ACOD1 protein levels (B). ****P < 0.0001 by unpaired, 2-tailed t test.(C) Production of ACOD1–/– macrophages. ipMDM-Ls and ipAM-Ls were differentiated from CRISPR-edited iPSCs (see Supplemental Figure 5) or isogenic control cells. Anti-ACOD1 immunoblot of ipMDM-L and ipAM-L cells derived from ACOD1-deficient iPSCs or isogenic controls stimulated with LPS as in A and B. (D) Human macrophage production of itaconate. GC-MS quantification of itaconate produced by mouse BMDMs, ipMDM-Ls, ipAM-Ls, or ACOD1–/– ipMDM-Ls and ipAM-Ls without (–) or with (+) 6 hours of LPS stimulation. Itaconate levels are graphed AU by normalization to total protein levels; 2-way ANOVA used to assess variance of the samples within the same treatment group and based on stimulation status. ****P < 0.0001. (E) Gene counts of ACOD1 mRNA after 4 hours or 24 hours without (–) or with (+) Mtb infection of ipMDM-Ls or ipAM-Ls. Data derived from 3 donors in biological duplicates. ***P < 0.005 ****P < 0.0001 by 2-way ANOVA. (F) Anti-ACOD1/Vinculin immunoblot of WT ipMDM-L cells after no treatment (NT), LPS stimulation (100 ng/mL), or Mtb infection at MOI of 3 and assayed at days 1 and 2 (top), or days 0, 3, and 5 (bottom). (G) Anti-ACOD1/Vinculin immunoblot of WT ipMDM-L cells after no treatment (NT), 4 hours of LPS stimulation (100 ng/mL), or infection with Mtb at MOI of 10 for 4 hours, or both. (H) GC-MS quantification of itaconate produced by ipMDM-Ls and ipAM-Ls after 4 hours of infection with Mtb (MOI 10) or LPS (100 ng/mL). Itaconate levels were normalized to total protein levels.