Abstract
Experimental autoimmune encephalomyelitis (EAE) is a mouse model of multiple sclerosis (MS) in which Th17 cells have a crucial but unclear function. Here we show that choline acetyltransferase (ChAT), which synthesizes acetylcholine (ACh), is a critical driver of pathogenicity in EAE. Mice with ChAT-deficient Th17 cells resist disease progression and show reduced brain-infiltrating immune cells. ChAT expression in Th17 cells is linked to strong TCR signaling, expression of the transcription factor Bhlhe40, and increased Il2, Il17, Il22, and Il23r mRNA levels. ChAT expression in Th17 cells is independent of IL21r signaling but dampened by TGFβ, implicating ChAT in controlling the dichotomous nature of Th17 cells. Our study establishes a cholinergic program in which ACh signaling primes chronic activation of Th17 cells, and thereby constitutes a pathogenic determinant of EAE. Our work may point to novel targets for therapeutic immunomodulation in MS.
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Data availability
The bulk RNA-seq and the ATAC-seq raw and processed data files can be accessed using the GEO SuperSeries with accession GSE215673. The Bhlhe40 TFBS track shown in Fig. S2C is from the JASPAR CORE 2022 Predicted Transcription Factor Binding Sites repository incorporated as a standard track into the UCSC Genome Browser. The Bhlhe40 ChIPseq track shown in Fig. S2E corresponds to the IgG-rab ChIP-seq Signal from the Snyder Lab dataset (ENCODE/SYDH, GSM935440). In addition, all data needed to evaluate the conclusions in this publication are present in the main article or the Supplementary Information.
Code availability
Critical scripts used to analyze the RNAseq and ATACseq data presented in the paper are available at https://github.com/CodeInTheSkies/Th17-Cell-Pathogenicity-Data.
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Acknowledgements
We thank the Princess Margaret Genomics Centre and the UHN Bioinformatics and HPC Core for RNA sequencing; the Weill Cornell Medicine and Genome Technology Center (GTC) at NYU for ATAC sequencing; and the Pathology Core at The Centre for Phenogenomics in Toronto for all histology services, antibody optimization, and immunohistochemistry. We are grateful to M.E.S. for scientific editing and Dr. Thorsten Berger for careful reading of the manuscript. We also thank the Genotyping Facility and the Animal Resource Centre at the Princess Margaret Cancer Centre (Toronto).
Funding
This work was supported by a grant to T.W.M. and R.N. from the Canadian Institutes of Health Research (CIHR) and a postdoctoral fellowship to R.N. from EMBO and Marie Curie (ALTF 725-2015).
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R.N. and T.W.M conceived the project. R.N. and D.N. initiated the project, designed and performed experiments, and analyzed data. G.S.D., C.Z., C.G., V.B.C., and M.G. performed experiments and analyzed data. K.T.G., J.H., A.C.W., and B.S. assisted with experiments. P.R. and Z.L. performed bioinformatic analyses. R.N. wrote the original draft of the manuscript with input from D.N., P.R., M.E.S., and T.W.M. R.A.F. kindly provided Il17a-Katushka mice. T.W.M supervised the research. All authors have reviewed and approved the manuscript in its current form.
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The authors do not have competing financial interests concerning the work described. T. Mak owns equity in Treadwell Therapeutics Inc. and Agios Pharmaceuticals and is a consultant for AstraZeneca and Tessa Therapeutics.
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Nechanitzky, R., Nechanitzky, D., Ramachandran, P. et al. Cholinergic control of Th17 cell pathogenicity in experimental autoimmune encephalomyelitis. Cell Death Differ 30, 407–416 (2023). https://doi.org/10.1038/s41418-022-01092-y
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DOI: https://doi.org/10.1038/s41418-022-01092-y
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