Graft-versus-host disease of the CNS is mediated by TNF upregulation in microglia
Nimitha R. Mathew, Janaki M. Vinnakota, Petya Apostolova, Daniel Erny, Shaimaa Hamarsheh, Geoffroy Andrieux, Jung-Seok Kim, Kathrin Hanke, Tobias Goldmann, Louise Chappell-Maor, Nadia El-Khawanky, Gabriele Ihorst, Dominik Schmidt, Justus Duyster, Jürgen Finke, Thomas Blank, Melanie Boerries, Bruce R. Blazar, Steffen Jung, Marco Prinz, Robert Zeiser
Nimitha R. Mathew, Janaki M. Vinnakota, Petya Apostolova, Daniel Erny, Shaimaa Hamarsheh, Geoffroy Andrieux, Jung-Seok Kim, Kathrin Hanke, Tobias Goldmann, Louise Chappell-Maor, Nadia El-Khawanky, Gabriele Ihorst, Dominik Schmidt, Justus Duyster, Jürgen Finke, Thomas Blank, Melanie Boerries, Bruce R. Blazar, Steffen Jung, Marco Prinz, Robert Zeiser
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Research Article

Graft-versus-host disease of the CNS is mediated by TNF upregulation in microglia

Abstract

Acute graft-versus-host disease (GVHD) can affect the central nervous system (CNS). The role of microglia in CNS-GVHD remains undefined. In agreement with microglia activation, we found that profound morphological changes and MHC-II and CD80 upregulation occurred upon GVHD induction. RNA sequencing–based analysis of purified microglia obtained from mice with CNS-GVHD revealed TNF upregulation. Selective TNF gene deletion in microglia of Cx3cr1creER Tnffl/– mice reduced MHC-II expression and decreased CNS T cell infiltrates and VCAM-1+ endothelial cells. GVHD increased microglia TGF-β–activated kinase-1 (TAK1) activation and NF-κB/p38 MAPK signaling. Selective Tak1 deletion in microglia using Cx3cr1creER Tak1fl/fl mice resulted in reduced TNF production and microglial MHC-II and improved neurocognitive activity. Pharmacological TAK1 inhibition reduced TNF production and MHC-II expression by microglia, Th1 and Th17 T cell infiltrates, and VCAM-1+ endothelial cells and improved neurocognitive activity, without blocking graft-versus-leukemia effects. Consistent with these findings in mice, we observed increased activation and TNF production of microglia in the CNS of GVHD patients. In summary, we prove a role for microglia in CNS-GVHD, identify the TAK1/TNF/MHC-II axis as a mediator of CNS-GVHD, and provide a TAK1 inhibitor–based approach against GVHD-induced neurotoxicity.

Authors

Nimitha R. Mathew, Janaki M. Vinnakota, Petya Apostolova, Daniel Erny, Shaimaa Hamarsheh, Geoffroy Andrieux, Jung-Seok Kim, Kathrin Hanke, Tobias Goldmann, Louise Chappell-Maor, Nadia El-Khawanky, Gabriele Ihorst, Dominik Schmidt, Justus Duyster, Jürgen Finke, Thomas Blank, Melanie Boerries, Bruce R. Blazar, Steffen Jung, Marco Prinz, Robert Zeiser

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Figure 7

VCAM-1 and ICAM-1 expression after allo-HCT declines upon TAK1 inhibition.

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VCAM-1 and ICAM-1 expression after allo-HCT declines upon TAK1 inhibitio...
(A–D) Representative flow cytometry plots and the respective cumulative scatter plots showing quantification of the fold change of MFI of VCAM-1 (A and B) and ICAM-1 (C and D) expression in endothelial cells (CD31+CD105+) from the CNS of BALB/c mice treated with vehicle (n = 6) or takinib (n = 7) isolated on day 14 after allo-HCT. The experiment was performed once. (E–G) Immunofluorescence staining and scatter plots indicating the percentage of brain CD34+ endothelial cells expressing VCAM-1 and DAPI derived from BALB/c mice treated with vehicle (n = 7) or takinib (n = 7) (E) and from Tnffl/– (n = 7) or Cx3cr1creER Tnffl/– (n = 6) mice (F) on day 14 after allo-HCT. (G) Representative images from Tnffl/– and Cx3cr1creER Tnffl/– mice, respectively, are shown. Scale bars: 50 μm. The experiment was performed once. (H) Scatter plot showing the percentage of open-arm entries by mice treated with vehicle (n = 11), takinib (n = 12), or 5-Oz (n = 12) in an elevated plus maze test. (I) Scatter plot showing the percentage of time spent by mice treated with vehicle (n = 13), takinib (n = 12), or 5-Oz (n = 12) in exploring a novel object in a novel object recognition test. The experiments were performed 3 times, and the results (mean ± SEM) were pooled. (J) Survival rates of C57BL/6 mice with transplanted AML (FLT3-ITD/MLL-PTD) cells and BALB/c (WT) bone marrow (BM) along with (white and blue circles) and without (black circles) allogeneic T cells. (K) Survival rates of BALB/c mice with transplanted AML (WEHI-3B) cells and C57BL/6 BM (WT) along with (white and blue circles) and without (black circles) allogeneic T cells. The experiments were performed twice, and the results were pooled. P values were calculated using 2-sided Student’s unpaired t test (B, D, and F), Mann-Whitney U test (E), 1-way ANOVA (H and I), or 2-sided Mantel-Cox test (J and K).